1 00:00:05,240 --> 00:00:08,560 WELCOME BACK TO DAY 2 OF THE 2 00:00:08,560 --> 00:00:09,040 THIS FANTASTIC MEETING. 3 00:00:09,040 --> 00:00:13,560 IT'S MY PLEASURE TO INTRODUCE 4 00:00:13,560 --> 00:00:15,080 DR. SONG WHO IS OUR FIRST 5 00:00:15,080 --> 00:00:29,640 SPEAKER TODAY FOR A SHORT TALK. 6 00:00:29,640 --> 00:00:29,880 WELCOME. 7 00:00:29,880 --> 00:00:31,360 >> THANK YOU. 8 00:00:31,360 --> 00:00:32,760 THANK YOU NIKI, THANK YOU PAM 9 00:00:32,760 --> 00:00:38,640 FOR DOING THESE FOR ALL THESE 10 00:00:38,640 --> 00:00:38,960 YEARS. 11 00:00:38,960 --> 00:00:47,120 DID I HEAR AN ECHO? 12 00:00:47,120 --> 00:00:48,600 ALL RIGHT. 13 00:00:48,600 --> 00:00:49,160 OKAY, RIGHT HERE. 14 00:00:49,160 --> 00:00:54,680 ALL RIGHT, SO I'M GOING TO--IT'S 15 00:00:54,680 --> 00:00:55,000 NOT--OKAY. 16 00:00:55,000 --> 00:00:59,720 SO I'M GOING TO TALK ABOUT THE 17 00:00:59,720 --> 00:01:01,360 GRANT WE WERE AWARDED AND HAVE 18 00:01:01,360 --> 00:01:03,560 BYE-BYE WORKING ON FOR SEVERAL 19 00:01:03,560 --> 00:01:14,080 YEARS, SO IT'S A FOSSIL CHEMICAL 20 00:01:14,080 --> 00:01:18,360 METHOD TO AN ISOLATE GLYCAN TO 21 00:01:18,360 --> 00:01:20,240 ADDRESS THE LACK OF GLYCANS IN A 22 00:01:20,240 --> 00:01:21,760 DIFFERENT WAY, SO I DON'T HAVE 23 00:01:21,760 --> 00:01:23,760 TO SHOW THESE FOR THE WHOLE 24 00:01:23,760 --> 00:01:25,480 AUDIENCE BUT THERE ARE MANY, 25 00:01:25,480 --> 00:01:27,760 MANY DIFFERENT KIND OF GLYCANS 26 00:01:27,760 --> 00:01:32,080 BUT THEY OCCUR MOSTLY AS THE 27 00:01:32,080 --> 00:01:34,360 GLYCAN CONJUGATES INSTEAD OF 28 00:01:34,360 --> 00:01:36,360 FREE PRODUCING GLYCANS. 29 00:01:36,360 --> 00:01:38,600 SO IF WE--OH, SORRY. 30 00:01:38,600 --> 00:01:39,000 THIS 1. 31 00:01:39,000 --> 00:01:41,800 SO WE ALL AGREE, AT LEAST IN 32 00:01:41,800 --> 00:01:43,840 THIS COMMUNITY THAT 1 OF THE 33 00:01:43,840 --> 00:01:46,960 MAJOR BARRIER IF IT'S NOT THE 34 00:01:46,960 --> 00:01:49,160 ONLY 1, BUT THE 1 OF THE MAJOR 35 00:01:49,160 --> 00:02:03,400 BARRIER IS THE LACK OF--OH. 36 00:02:03,400 --> 00:02:04,040 READILY RELEVANT CARBOHYDRATES. 37 00:02:04,040 --> 00:02:05,680 SO HOW DO WE GET THESE? 38 00:02:05,680 --> 00:02:07,360 THE COMMUNITY HAS BEEN WORKING 39 00:02:07,360 --> 00:02:09,160 ON THESE VERY DILIGENTLY, SO 40 00:02:09,160 --> 00:02:10,560 THESE ARE RELEVANT DPLI 41 00:02:10,560 --> 00:02:13,600 KAN--KANAS, IT'S BASICALLY 2 42 00:02:13,600 --> 00:02:14,680 WAYS, EITHER THROUGH VETTING 43 00:02:14,680 --> 00:02:17,360 POSSIBLE, OR THROUGH THE NATURAL 44 00:02:17,360 --> 00:02:21,920 PRODUCTS ISOLATION, SEPARATION. 45 00:02:21,920 --> 00:02:30,760 SO, FOR--I AM GOING TO USE THE 46 00:02:30,760 --> 00:02:32,120 OTHER 1. 47 00:02:32,120 --> 00:02:34,160 YEAH, SO CHEMICAL, HAVE YOU 48 00:02:34,160 --> 00:02:37,320 HEARD YESTERDAY, A VERY 49 00:02:37,320 --> 00:02:41,160 IMPRESSIVE TALKS FROM 50 00:02:41,160 --> 00:02:43,200 DR. WILSON, AND DR. JAN LUOT 51 00:02:43,200 --> 00:02:44,800 SYNTHESIS OF IMMUNO GLYCANS, BUT 52 00:02:44,800 --> 00:02:48,760 YOU WILL HEAR A LOT MORE ON 53 00:02:48,760 --> 00:02:52,480 THE--SYNTHESIS OF OTHER CLASS OF 54 00:02:52,480 --> 00:02:54,360 GLYCANS, BUT WHAT WE'RE GOING TO 55 00:02:54,360 --> 00:02:57,240 FOCUS ON IS THE WAY TO ISOLATE 56 00:02:57,240 --> 00:03:00,760 AND SEPARATE THE NATURAL 57 00:03:00,760 --> 00:03:03,760 GLYCANS. 58 00:03:03,760 --> 00:03:09,880 SO AS WE--OKAY, SO CAN WE REALLY 59 00:03:09,880 --> 00:03:11,840 PRODUCE LARGER AMOUNT OF GLYCAN 60 00:03:11,840 --> 00:03:13,840 FROM MEASURABLE SOURCES, SO UP 61 00:03:13,840 --> 00:03:18,640 UNTIL NOW, THERE'S PRODUCING 62 00:03:18,640 --> 00:03:20,160 NATURAL GLYCANS FOR REASON HAS 63 00:03:20,160 --> 00:03:22,320 BEEN IN THE GRIEK O BIOLOGY AREA 64 00:03:22,320 --> 00:03:25,360 FOR AT LEAST AS LONG AS 65 00:03:25,360 --> 00:03:26,720 SYNTHESIS, SO, IT'S--IT'S NOT A 66 00:03:26,720 --> 00:03:29,520 PROBLEM TO GET A SMALL AMOUNT OF 67 00:03:29,520 --> 00:03:31,160 A GLYCAN. 68 00:03:31,160 --> 00:03:34,040 WE HAVE ENZYMES, WE HAVE SOME 69 00:03:34,040 --> 00:03:36,720 CHEMICALS THAT CAN BE USED TO 70 00:03:36,720 --> 00:03:43,680 RELEASE GLYCANS, BUT THERE ARE 71 00:03:43,680 --> 00:03:46,560 SEVERAL PROBLEM OF THESE--EXCUSE 72 00:03:46,560 --> 00:03:48,480 ME,--VERY SENSITIVE. 73 00:03:48,480 --> 00:03:50,960 SO THE LOW AMOUNT OF NATURAL 74 00:03:50,960 --> 00:03:53,320 MATERIALS, AND THERE'S NO 75 00:03:53,320 --> 00:03:54,600 SCALABLE PROCESS TO RELEASE GLUE 76 00:03:54,600 --> 00:03:56,360 MARIOUS ANS AND THE GLYCANS 77 00:03:56,360 --> 00:03:59,360 WHATEVER YOU RELEASE IS A 78 00:03:59,360 --> 00:04:00,600 HETEROGENIUS DPLI KAN--KANA MIX 79 00:04:00,600 --> 00:04:01,440 TOUR THAT REQUIRES SEPARATION, 80 00:04:01,440 --> 00:04:04,320 SO WHAT WE WANT TO ADDRESS IS 81 00:04:04,320 --> 00:04:14,360 WEATHER WE CAN ACTUALLY LOOK AT 82 00:04:14,360 --> 00:04:17,360 AND TO DEAL WITH A LARGER AMOUNT 83 00:04:17,360 --> 00:04:18,760 OF NATURAL MATERIAL SO WE CAN 84 00:04:18,760 --> 00:04:20,840 GET LARGER AMOUNT OF NATURAL GUY 85 00:04:20,840 --> 00:04:22,760 KAN--KANAS SO A FEW YEARS AGO, I 86 00:04:22,760 --> 00:04:28,760 NOTICED THIS PAPER IF YOU HAVE 87 00:04:28,760 --> 00:04:30,120 YOU CANOXIDATE THIS TO A TRIAL. 88 00:04:30,120 --> 00:04:31,960 SO WE'RE THINKING IF YOU HAVE A 89 00:04:31,960 --> 00:04:40,560 GLYCO PROTEIN, WE KNOW PRONAIS 90 00:04:40,560 --> 00:04:43,080 CAN TO GET THE LAST GLYCO IMMUNO 91 00:04:43,080 --> 00:05:08,000 ASSAY, SO WHAT WE WILL GET IF WE 92 00:05:08,000 --> 00:05:09,120 ORIGINAL REDUCING GLYCANS IF YOU 93 00:05:09,120 --> 00:05:10,760 HEATED THE MIXTURE UP, BUT THIS 94 00:05:10,760 --> 00:05:13,800 METHOD IS NOT IDEAL BECAUSE IT'S 95 00:05:13,800 --> 00:05:15,080 STILL INVOLVED ENZYME IS IT'S 96 00:05:15,080 --> 00:05:16,280 REALLY YOU CAN IMAGINE DOING 97 00:05:16,280 --> 00:05:20,480 THIS WITH A HUNDRED GRAMS OF 98 00:05:20,480 --> 00:05:22,360 MATERIAL MATERIAL BUT YOU CANNOT 99 00:05:22,360 --> 00:05:23,360 WORK WITH KILOGRAMS OF MATERIAL 100 00:05:23,360 --> 00:05:27,200 AND ALSO THIS THING IS STILL 101 00:05:27,200 --> 00:05:28,840 CHEMICAL BY ITSELF DOESN'T 102 00:05:28,840 --> 00:05:30,920 WELCOMER ON LIKE UNTREATED 103 00:05:30,920 --> 00:05:32,640 MATERIAL AND ALSO IT DOESN'T 104 00:05:32,640 --> 00:05:35,000 WORK ON O-GLUE MARIOUS ANS AND 105 00:05:35,000 --> 00:05:36,760 THE LIPIDS SO WE WERE LOOKING 106 00:05:36,760 --> 00:05:40,000 BUT WE WERE ENCOURAGED BY 107 00:05:40,000 --> 00:05:41,160 THEOXIDATIVE DEVIATION PATHWAY, 108 00:05:41,160 --> 00:05:49,080 SO WE WERE LOOKING FOR OTHEROX 109 00:05:49,080 --> 00:05:50,280 DABTS, SO THE FIRST THING MEAN 110 00:05:50,280 --> 00:05:54,600 WE TRY, ACTUALLY WORKED WHICH IS 111 00:05:54,600 --> 00:05:55,760 SODIUM HYDROCHLORIDE, WHICH IS 112 00:05:55,760 --> 00:05:57,760 PROBABLY THE SIMPLEST INOX DACE 113 00:05:57,760 --> 00:06:00,160 IN EVERYONE'S HOME, IT'S THE 114 00:06:00,160 --> 00:06:00,960 HOUSEHOLD BLEACH. 115 00:06:00,960 --> 00:06:08,080 SO, YOU TREAT A GLYCO PROTEIN 116 00:06:08,080 --> 00:06:10,200 WITH BLEACH, IS THEY ARE 117 00:06:10,200 --> 00:06:11,640 RELEASED, THE PROTEINS DEGRADED 118 00:06:11,640 --> 00:06:13,680 AND THE PROTEINS MOVE ON THE 119 00:06:13,680 --> 00:06:16,360 PLATE AND THEY VIEW--IF YOU LOOK 120 00:06:16,360 --> 00:06:18,160 AT THE MASS SPEC, CAN YOU SEE A 121 00:06:18,160 --> 00:06:20,480 LOT OF REDUCING GLYCANS HERE AND 122 00:06:20,480 --> 00:06:23,560 OF COURSE, THE SCHEME IS LIKE 123 00:06:23,560 --> 00:06:24,880 THESE, FOR NGLYCAN PROTEINS THEN 124 00:06:24,880 --> 00:06:29,080 WHAT YOU GET IS A FREE REDUCING 125 00:06:29,080 --> 00:06:29,680 GLYCAN. 126 00:06:29,680 --> 00:06:31,160 THIS DIRECTLY TREATS THE GLYCO 127 00:06:31,160 --> 00:06:34,960 PRACTICES TEEN, WE DON'T NEED 128 00:06:34,960 --> 00:06:38,240 ANY PROTEIN TREATED WITH ANY 129 00:06:38,240 --> 00:06:39,360 ENZYME TO BREAK THIS APART. 130 00:06:39,360 --> 00:06:43,680 AND THIS ALSO WORKS WITH 131 00:06:43,680 --> 00:06:45,640 [INDISCERNIBLE] AS LONG AS YOU 132 00:06:45,640 --> 00:06:46,120 HOMOGINIZE IT. 133 00:06:46,120 --> 00:06:50,000 SO IT WILL ALSO WORK ON OTHER 134 00:06:50,000 --> 00:06:52,160 THAT WILLATE, AND ALL THE 135 00:06:52,160 --> 00:06:54,560 GLYCANS IT'S VERY--HAVE YOU A 136 00:06:54,560 --> 00:06:56,560 LITTLE BIT OVER OXIDATION, SOME 137 00:06:56,560 --> 00:06:58,360 SIDE PRODUCT CHSES I WILL 138 00:06:58,360 --> 00:06:59,520 ADDRESS LITTLE BIT BUT OVER ALL 139 00:06:59,520 --> 00:07:03,720 IT HAS A VERY SIMILAR PROTILE BY 140 00:07:03,720 --> 00:07:05,560 BLEACH OR BY UPON 141 00:07:05,560 --> 00:07:07,080 [INDISCERNIBLE]. 142 00:07:07,080 --> 00:07:10,520 SO THE HPOC CONJUGATION, 143 00:07:10,520 --> 00:07:11,360 FLUORESCENT CONJUSTICE 144 00:07:11,360 --> 00:07:12,960 ACCESSICATION SHOWED THE MAJOR 145 00:07:12,960 --> 00:07:14,960 GLYCANS ARE RELEASED THE SAME BY 146 00:07:14,960 --> 00:07:16,960 BLEACH AND THE PGSF, THIS IS NOT 147 00:07:16,960 --> 00:07:24,440 AS CLEAN AS PGSF, BUT THAT'S 148 00:07:24,440 --> 00:07:26,560 SORT OF EXPECTED AND THE RELEASE 149 00:07:26,560 --> 00:07:28,520 OF THOSE GLYCANS ARE SPHRUCTURAL 150 00:07:28,520 --> 00:07:30,600 INCREASE ARE REALLY VERY WELL 151 00:07:30,600 --> 00:07:33,720 RESERVED SO IF WE PRINT A 152 00:07:33,720 --> 00:07:35,000 MICROARRAY, THE BINDING OF THE 153 00:07:35,000 --> 00:07:38,840 PROTEINS THEY ARE EXPECTED TO, 154 00:07:38,840 --> 00:07:39,600 SO ... THE GOOD THING ABOUT 155 00:07:39,600 --> 00:07:42,240 THESE IS THAT NOW WE CAN WORK 156 00:07:42,240 --> 00:07:45,640 WITH LARGER SCALE TO REALLY 157 00:07:45,640 --> 00:07:49,360 TARGET SOME HIGH MILLIGRAM 158 00:07:49,360 --> 00:07:50,920 COASTAL GRAM SCALE GLYCAN 159 00:07:50,920 --> 00:07:54,080 SEPARATION, THIS IS AN EXAMPLE 160 00:07:54,080 --> 00:07:55,960 THAT WE PURIFY GLYCANS AND THE 161 00:07:55,960 --> 00:07:58,680 GLYCANS LIKE THESE ON THEIR 162 00:07:58,680 --> 00:08:02,720 CURRENT MARKET, YOU'RE LOOKING 163 00:08:02,720 --> 00:08:04,440 AT LIKE A--LIKE 10 TO WENT 164 00:08:04,440 --> 00:08:05,880 MICROGRAM WITH SEVERAL HUNDRED 165 00:08:05,880 --> 00:08:07,560 DOLLARS PRICE THAT IS REALLY 166 00:08:07,560 --> 00:08:11,080 PREVENTING A LOT OF PEOPLE THAT 167 00:08:11,080 --> 00:08:15,480 GET INVOLVE WIDE STUDYING THEIR 168 00:08:15,480 --> 00:08:16,000 FUNCTIONS. 169 00:08:16,000 --> 00:08:17,840 SO AND ALSO WITH THESE AMOUNT OF 170 00:08:17,840 --> 00:08:21,160 MATERIAL, WE CAN GET A VERY NICE 171 00:08:21,160 --> 00:08:24,520 NMR DATA, BOTH PROTON MR AND 172 00:08:24,520 --> 00:08:27,200 CARBON 13 MR, SO FROM ABOUT A 173 00:08:27,200 --> 00:08:29,080 KILOGRAM OF SOY PROTEIN ISOLATES 174 00:08:29,080 --> 00:08:30,160 AND THIS IS APPROXIMATE 175 00:08:30,160 --> 00:08:32,360 SOMETHING PEOPLE TAKE AS A FULL 176 00:08:32,360 --> 00:08:35,760 SUPPLEMENT AND YOU CAN GET THE 177 00:08:35,760 --> 00:08:37,960 LIKE A HUNDRED MILLIGRAMS OF 178 00:08:37,960 --> 00:08:40,280 DIFFERENT STRUCTURE AND IF YOU 179 00:08:40,280 --> 00:08:42,200 NOTICE THERE ARE MONODPLIEWTEN 180 00:08:42,200 --> 00:08:51,680 STRUCTURES THAT LOSE IN THE 181 00:08:51,680 --> 00:08:53,000 TERMINAL GLUNAC, SO THIS IS 182 00:08:53,000 --> 00:08:54,640 INTACT THIS, IS GLYCANS RELEASED 183 00:08:54,640 --> 00:09:00,160 BY BLEACH BUT WE ALSO SEE SOME 184 00:09:00,160 --> 00:09:02,000 OTHER FOR THE TRIAL, YOU CAN 185 00:09:02,000 --> 00:09:02,600 ACTUALLY TRANSFORM THIS 186 00:09:02,600 --> 00:09:04,600 APPROXIMATE BACK TO REDUCING 187 00:09:04,600 --> 00:09:07,200 GLYCANS BY CREATING CARBON. 188 00:09:07,200 --> 00:09:08,040 ALSO ANOTHER VERY INTERESTING 189 00:09:08,040 --> 00:09:10,600 SIDE PRODUCT IS THIS 1 THAT IS 190 00:09:10,600 --> 00:09:13,600 LOSING THE TERMINAL GLUNAC IS A 191 00:09:13,600 --> 00:09:14,720 GOOD SUBSTRATE FOR THE MEDIATED 192 00:09:14,720 --> 00:09:32,720 AND FLIEK O PEPTIDE SYNTHESIS. 193 00:09:32,720 --> 00:09:35,760 WHETHER THIS CAN DO A RELEASE 194 00:09:35,760 --> 00:09:38,000 FOR GLYCO CONJUGATES AND WORK ON 195 00:09:38,000 --> 00:09:38,920 OGLYCAN SHOWN HERE AND THE 196 00:09:38,920 --> 00:09:41,200 IMPORTANT THING IS THE REDUCING 197 00:09:41,200 --> 00:09:43,560 THESE A LITTLE BIT DIFFERENT. 198 00:09:43,560 --> 00:09:45,800 SO THE THESE ARE STABLE TOWARDS 199 00:09:45,800 --> 00:09:46,040 BLEACH. 200 00:09:46,040 --> 00:09:48,560 SO YOU OX DIET THE PROTEIN ALL 201 00:09:48,560 --> 00:09:52,560 THE WAY TO THESE ASSAY, AND THEN 202 00:09:52,560 --> 00:09:54,360 AS STRONG AS YOU THINK PLEACH 203 00:09:54,360 --> 00:09:56,840 MIGHT BE HARSH TO THE 204 00:09:56,840 --> 00:09:57,920 STRUCTURES, IT ACTUALLY VERY 205 00:09:57,920 --> 00:10:01,320 MILD UNDER THIS CONDITIONS, 206 00:10:01,320 --> 00:10:03,040 BECAUSE EVEN [INDISCERNIBLE] 207 00:10:03,040 --> 00:10:04,440 GROUP ON ACID GOT PRESERVED VERY 208 00:10:04,440 --> 00:10:08,400 WELL IF YOU TRADED WITH LIMITED 209 00:10:08,400 --> 00:10:14,440 AMOUNT OF BLEACH. 210 00:10:14,440 --> 00:10:17,000 SO, THIS IS EXPWRUOF THE 211 00:10:17,000 --> 00:10:23,920 DEMONSTRATION OF WORKING ON 212 00:10:23,920 --> 00:10:24,560 O-GLYCANS FROM [INDISCERNIBLE]. 213 00:10:24,560 --> 00:10:26,880 SO A LOT OF THESE HAVE LIPID 214 00:10:26,880 --> 00:10:33,560 SAYS AND UNFORTUNATELY THIS IS 215 00:10:33,560 --> 00:10:35,560 ALSO WORKED ONLET PRODUCT AGAIN 216 00:10:35,560 --> 00:10:39,080 IS LIPID, AND IS STABLE, SO THIS 217 00:10:39,080 --> 00:10:47,200 PLEACH OXIDIZE THE WHOLE SERIES 218 00:10:47,200 --> 00:10:53,680 POINTS MID PART AND THIS IS SOME 219 00:10:53,680 --> 00:10:54,600 DEMONSTRATION THAT WILL COME, 220 00:10:54,600 --> 00:10:56,720 AND CAN BE SIZED. 221 00:10:56,720 --> 00:11:01,280 THE GOOD THING ABOUT THIS IS 222 00:11:01,280 --> 00:11:03,240 THAT IT'S NOT ONLY PURIFIED, 223 00:11:03,240 --> 00:11:10,000 THIS BUT THIS WORKED DIRECTLY ON 224 00:11:10,000 --> 00:11:11,080 [INDISCERNIBLE] AND THEN YOU 225 00:11:11,080 --> 00:11:13,160 NEED TO ISOLATE THEM FROM THE 226 00:11:13,160 --> 00:11:15,680 MIX TOUR AND TREATED CHEMICALLY, 227 00:11:15,680 --> 00:11:17,960 GLYCANS ARE VERY DIFFERENT FROM 228 00:11:17,960 --> 00:11:20,040 OTHER BOWEL MOLECULES, AND SO 229 00:11:20,040 --> 00:11:21,080 IT'S--YOU CAN FIND DIFFERENT 230 00:11:21,080 --> 00:11:22,040 WAYS TO SEPARATE THEM. 231 00:11:22,040 --> 00:11:26,040 SO AS A SUMMARY, WE WERE ABLE TO 232 00:11:26,040 --> 00:11:29,200 USE THE SINGLE BLEACH OR 233 00:11:29,200 --> 00:11:30,600 REAGENT, UNDER VERY MILD 234 00:11:30,600 --> 00:11:31,680 CONDITION TO REDEC THESE 3 235 00:11:31,680 --> 00:11:33,680 CLASSES OF DPLI KAN--KANAS. 236 00:11:33,680 --> 00:11:35,880 AND THE MECHANISM, I WOULD SAY 237 00:11:35,880 --> 00:11:36,840 MECHANISM IS PROBABLY MORE 238 00:11:36,840 --> 00:11:38,360 COMPLICATED THAN I SHOW HERE, 239 00:11:38,360 --> 00:11:46,760 AND AS WE SEE A DIFFERENT AS WE 240 00:11:46,760 --> 00:11:49,000 SEE A MORE SUBTLE SIDE, 241 00:11:49,000 --> 00:11:49,960 HOPEFULLY WE WILL REPORT LATER 242 00:11:49,960 --> 00:11:53,560 BUT WE WERE THINKING IT MOSTLY 243 00:11:53,560 --> 00:11:56,160 PROBABLY FROM THIS CLARINATION 244 00:11:56,160 --> 00:12:00,400 INITIATED DEGRADATION IMMUNATION 245 00:12:00,400 --> 00:12:01,760 ON REACTION. 246 00:12:01,760 --> 00:12:04,240 SO JUST THE CHALLENGES COMPARED 247 00:12:04,240 --> 00:12:08,520 TO PREVIOUS METHODS, SO IT'S A 248 00:12:08,520 --> 00:12:10,520 CHEAP REGIONS, SIMPLE OPERATION 249 00:12:10,520 --> 00:12:13,160 AND DIRECTLY APICKABLE TO ANIMAL 250 00:12:13,160 --> 00:12:14,440 PLANT TISSUES INTO HOMOGINIZE 251 00:12:14,440 --> 00:12:17,280 IT, YOU DON'T NEED PREEXTRACTION 252 00:12:17,280 --> 00:12:17,960 OF GLYCO CONJUSTICE ACCESS GAYS, 253 00:12:17,960 --> 00:12:20,560 AND THAT IS THE LARGE SCALE, IS 254 00:12:20,560 --> 00:12:20,800 POSSIBLE. 255 00:12:20,800 --> 00:12:24,160 SO WE ARE WORKING WITH KILOGRAM 256 00:12:24,160 --> 00:12:26,480 STUFF AND ALL YOU NEED IS THE 257 00:12:26,480 --> 00:12:28,840 OVERHEAD AND THE MECHANICAL 258 00:12:28,840 --> 00:12:31,360 STIRRER AND THAT IS THE REACTION 259 00:12:31,360 --> 00:12:33,920 MIXTURE, IT IS AN AQUEOUS 260 00:12:33,920 --> 00:12:34,960 SOLUTION, AND THE SOLUTION GETS 261 00:12:34,960 --> 00:12:37,480 WARM A LITTLE BIT BUT IT'S 262 00:12:37,480 --> 00:12:41,600 NOTHING TO INTERESTING LIKE 263 00:12:41,600 --> 00:12:44,200 [INDISCERNIBLE] THE SCALE THAT 264 00:12:44,200 --> 00:12:46,560 WE TRIED, A HUNDRED LITERS SO 265 00:12:46,560 --> 00:12:48,720 THE DISADVANTAGE, SO THE 266 00:12:48,720 --> 00:12:51,360 CHALLENGE, IT'S NOT AS CLEAN AS 267 00:12:51,360 --> 00:12:52,360 ENZYMATIC CLEAVAGE BUT THIS 268 00:12:52,360 --> 00:12:54,120 STAGE IS MOSTLY USED FOR 269 00:12:54,120 --> 00:12:55,640 PREPARED SCALE AND REACTIONS AND 270 00:12:55,640 --> 00:12:56,960 MEASUREMENTS, I THINK WE ARE 271 00:12:56,960 --> 00:12:58,400 GETTING CLOSER AND CLOSER TO 272 00:12:58,400 --> 00:13:00,120 UNDERSTAND THE OLD MECHANISMS 273 00:13:00,120 --> 00:13:03,080 THROUGH THE ISOLATION OF THESE 274 00:13:03,080 --> 00:13:04,360 SITE PRODUCT BUT WE ARE STILL 275 00:13:04,360 --> 00:13:06,440 WORKING ON IT AND THERE'S 276 00:13:06,440 --> 00:13:07,160 OVEROXIDATION SIDE PRODUCT BUT 277 00:13:07,160 --> 00:13:13,560 SOME OF THEM COULD BE VERY 278 00:13:13,560 --> 00:13:15,760 USEFUL FOR SOME DIFFERENT 279 00:13:15,760 --> 00:13:16,360 OPERATING GLOBALLYLICATIONS, 280 00:13:16,360 --> 00:13:18,360 THIS IS DIFFICULT BECAUSE THERE 281 00:13:18,360 --> 00:13:19,560 ARE MIXTURES TOO REQUIRE 282 00:13:19,560 --> 00:13:20,240 SEPARATION. 283 00:13:20,240 --> 00:13:23,560 WE HAVE MADE SIGNIFICANT 284 00:13:23,560 --> 00:13:26,760 ADVANCES OVER HERE BUT I'M NOW 285 00:13:26,760 --> 00:13:28,680 GOING TO TALK ABOUT THAT TODAY 286 00:13:28,680 --> 00:13:29,320 FOR TIME'S SAKE. 287 00:13:29,320 --> 00:13:34,920 SO IN SORT WE WERE ABLE TO LIKE 288 00:13:34,920 --> 00:13:36,960 BRING THE ISOLATION OF NATURAL 289 00:13:36,960 --> 00:13:38,440 TBLI KAN--KANAS FROM NATURAL 290 00:13:38,440 --> 00:13:41,040 MATERIAL TO THE NEXT LEVEL. 291 00:13:41,040 --> 00:13:45,040 AND REALLY IT BECOMES 292 00:13:45,040 --> 00:13:45,760 ALTERNATIVE [INDISCERNIBLE] 293 00:13:45,760 --> 00:13:46,720 TOWARD COMPLEX GLYCANS SO FOR 294 00:13:46,720 --> 00:13:50,040 NOW, IF YOU ARE WORKING ON LIKE 295 00:13:50,040 --> 00:13:51,200 SOME [INDISCERNIBLE] DPLI 296 00:13:51,200 --> 00:13:53,400 KAN--KANAS, TELL BE VERY HARD 297 00:13:53,400 --> 00:13:55,720 FOR YOU WANT TO DO A SYNTHESIS 298 00:13:55,720 --> 00:13:56,560 EXIST AT THIS POINT. 299 00:13:56,560 --> 00:14:34,000 BUT NOT FOR THOSE MINOR GLYCANS. 300 00:14:34,000 --> 00:14:35,160 --WE HAD PUBLISHED A LITTLE 301 00:14:35,160 --> 00:14:37,840 PAPER THAT USING NBS AGAIN, WE 302 00:14:37,840 --> 00:14:39,880 CAN REGENERATE THE REDUCING 303 00:14:39,880 --> 00:14:42,640 GLYCANS BUT THIS 1 LOOSING A 304 00:14:42,640 --> 00:14:44,360 CARBON IS THE MAJOR SIDE 305 00:14:44,360 --> 00:14:45,240 PRODUCT. 306 00:14:45,240 --> 00:14:46,600 SO MOST RECENTLY, WE ARE WORKING 307 00:14:46,600 --> 00:14:49,960 ON USING DIFFERENT REGIONS, 308 00:14:49,960 --> 00:14:52,680 USING OXOM, WHICH GIVE US MUCH 309 00:14:52,680 --> 00:14:56,080 LESS OF THE PENTOSE, FOR 310 00:14:56,080 --> 00:14:57,800 REDUCING GLYCANS BUT WE LIST 311 00:14:57,800 --> 00:15:02,520 SIDE PART, THIS CAN CONVERT BACK 312 00:15:02,520 --> 00:15:04,040 TO REDUCING [INDISCERNIBLE] BY 313 00:15:04,040 --> 00:15:04,400 REDUCING CARBON. 314 00:15:04,400 --> 00:15:06,200 SO WITH THIS I WOULD LIKE TO 315 00:15:06,200 --> 00:15:07,560 THANK EVERYONE, ESPECIALLY THANK 316 00:15:07,560 --> 00:15:09,840 THE COMMUNITY, THANK PAM, FOR 317 00:15:09,840 --> 00:15:12,240 PUTTING THIS TOGETHER AND THESE 318 00:15:12,240 --> 00:15:13,360 ARE THE PEOPLE WHO DO THE WORK. 319 00:15:13,360 --> 00:15:26,560 THANK YOU VERY MUCH. 320 00:15:26,560 --> 00:15:27,360 [ APPLAUSE ] 321 00:15:27,360 --> 00:15:28,200 >> THANK YOU VERY MUCH. 322 00:15:28,200 --> 00:15:30,080 MY LAB DEVELOPED THEOXIDATIVE 323 00:15:30,080 --> 00:15:31,760 PROCESS FOR THE FIRST YEAR, 324 00:15:31,760 --> 00:15:33,440 UNDERGRADUATE LAB CLASS AND WE 325 00:15:33,440 --> 00:15:34,960 JUST PUBLISHED THAT, AND SO IT'S 326 00:15:34,960 --> 00:15:35,920 STRAIGHT FORWARD ENOUGH THAT 327 00:15:35,920 --> 00:15:37,760 FRESHMAN CAN DO IT. 328 00:15:37,760 --> 00:15:42,560 SO OUR NEXT TALK IS VIRTUAL AND 329 00:15:42,560 --> 00:15:44,040 UNFORTUNATELY DR. DAVID SMITH 330 00:15:44,040 --> 00:15:45,360 COULD NOT BE WITH US HERE TODAY 331 00:15:45,360 --> 00:15:48,560 LIVE AND WE WILL HEAR FROM HIM 332 00:15:48,560 --> 00:15:49,520 VIRTUALLY. 333 00:15:49,520 --> 00:15:50,880 FIRST OF ALL I WANT TO THANK 334 00:15:50,880 --> 00:15:58,120 PAM, AND ALL THE ORGANIZERS OF 335 00:15:58,120 --> 00:16:01,320 THIS MEETING TO PROVIDE THEM A 336 00:16:01,320 --> 00:16:04,720 BRIEF SUMMARY OF THE TECHNOLOGY 337 00:16:04,720 --> 00:16:06,320 THAT WAS DEVELOPED AND YOU KNOW 338 00:16:06,320 --> 00:16:18,400 THIS WAS DEVELOPED WITH MYSELF 339 00:16:18,400 --> 00:16:20,160 AND [INDISCERNIBLE]. 340 00:16:20,160 --> 00:16:22,320 NATURAL GLYCANS REAGENTS AND 341 00:16:22,320 --> 00:16:23,600 STANDARDS PREUSING ORNG, AND WE 342 00:16:23,600 --> 00:16:26,320 JUST HEARD SOME DETAILS OF THIS 343 00:16:26,320 --> 00:16:30,160 BUT OUR MESSAGE TODAY FOR 344 00:16:30,160 --> 00:16:32,120 NGLYCAN, OF NATURAL NGLYCANS ARE 345 00:16:32,120 --> 00:16:34,080 AVAILABLE AND AFFORDABLE THANKS 346 00:16:34,080 --> 00:16:35,600 TO THE NIH COMMON FUND AND ALL 347 00:16:35,600 --> 00:16:39,720 YOU FOLKS INVOLVED IN MAKING 348 00:16:39,720 --> 00:16:41,160 THIS HAPPEN. 349 00:16:41,160 --> 00:16:42,360 IN HIS PRESENTATION, DR. SONG 350 00:16:42,360 --> 00:16:46,520 POINTED TO THE FACT THAT LACK OF 351 00:16:46,520 --> 00:16:48,280 LARGE GLYCAN LIBRARIES AND 352 00:16:48,280 --> 00:16:49,800 MILLIGRAM TO GRAM QUANTITIES 353 00:16:49,800 --> 00:16:51,680 LIMITS THEIR USE AS DRUG 354 00:16:51,680 --> 00:16:51,920 TARGETS. 355 00:16:51,920 --> 00:16:57,400 THIS IS TO SUPPORT THIS, I WANT 356 00:16:57,400 --> 00:17:00,840 TO USE OLIGO SACCHARIDES AS AN 357 00:17:00,840 --> 00:17:01,080 EXAMPLE. 358 00:17:01,080 --> 00:17:03,600 HMO IS AN ATTAINABLE FLUID WHICH 359 00:17:03,600 --> 00:17:07,200 IS RICH IN COMPLEX OLIGO 360 00:17:07,200 --> 00:17:08,120 SACCHARIDES, THESE WERE 361 00:17:08,120 --> 00:17:09,520 INTENSELY STUDIED IN THE 1940S 362 00:17:09,520 --> 00:17:11,880 AFTER IT WAS DETERMINE THAD MANY 363 00:17:11,880 --> 00:17:13,120 OF THESE OLIGO SACCHARIDE 364 00:17:13,120 --> 00:17:15,280 STRUCTURES SUPPORT THE GROUP OF 365 00:17:15,280 --> 00:17:18,040 DIFFICULT BACTERIA WHICH WAS A 366 00:17:18,040 --> 00:17:18,960 BENEFICIAL BACTERIA IN THE GUT 367 00:17:18,960 --> 00:17:21,720 OF PREOF THE FED INFANTS AND THE 368 00:17:21,720 --> 00:17:25,120 DATA BUILT THE QUANTITIES OF 369 00:17:25,120 --> 00:17:25,880 THESE OLIGO SACCHARIDES 370 00:17:25,880 --> 00:17:27,120 STIMULATED RESEARCH AND METHODS 371 00:17:27,120 --> 00:17:29,880 FOR SEPARATION AND ISOLATION OF 372 00:17:29,880 --> 00:17:30,320 OLIGO SACCHARIDES. 373 00:17:30,320 --> 00:17:31,880 IN THE EPPED IN THE DEFINITION 374 00:17:31,880 --> 00:17:33,920 OF DPLI KAN--KANA STRUCTURE 375 00:17:33,920 --> 00:17:35,240 SINCE YOU COULD PURIFY THEM AND 376 00:17:35,240 --> 00:17:36,880 THEY WERE AVAILABLE IN MILLIGRAM 377 00:17:36,880 --> 00:17:38,840 QUANTITIES AND MANY OF THEM 378 00:17:38,840 --> 00:17:40,480 STRUCTURAL METHODS BACK THEN 379 00:17:40,480 --> 00:17:42,440 REQUIRED MILLIGRAM QUANTITIES TO 380 00:17:42,440 --> 00:17:42,800 DO THIS WORK. 381 00:17:42,800 --> 00:17:45,200 ONCE THEY WERE DEFINED, THAT YOU 382 00:17:45,200 --> 00:17:47,600 WERE USED AS STANDARDS TO 383 00:17:47,600 --> 00:17:50,840 DEVELOP A MASS SPECTROMETRY, 384 00:17:50,840 --> 00:17:52,560 HELPED DEVELOP MASS 385 00:17:52,560 --> 00:17:54,600 SPECTROMETRY, AS GLYCAN FOR 386 00:17:54,600 --> 00:17:55,160 STRUCTURAL ANALYSIS. 387 00:17:55,160 --> 00:17:58,160 NOW, AS THE HMOs STRUCTURES 388 00:17:58,160 --> 00:17:59,720 BEHAIM DEFINED AND AVAILABLE, AS 389 00:17:59,720 --> 00:18:04,080 I SAID IN THE MILLIGRAM, THIS 390 00:18:04,080 --> 00:18:05,480 RESEARCH WAS CORRECTEDDA AT 391 00:18:05,480 --> 00:18:08,360 DEFINING THE FUNCTION WHICH 392 00:18:08,360 --> 00:18:09,600 INCLUDED PREBIOTIC ACTIVITY AND 393 00:18:09,600 --> 00:18:11,720 THEY ALSO SHOWED TO PROVIDE 394 00:18:11,720 --> 00:18:13,200 PROTECTION IN BREAST FAD 395 00:18:13,200 --> 00:18:15,240 INSTANTS FROM VARIOUS VIRUSES 396 00:18:15,240 --> 00:18:16,960 LIKE INFLUENZA AND ROTOVIRUS, 397 00:18:16,960 --> 00:18:25,160 BASED ON THEIR RECEPTOR DECOY 398 00:18:25,160 --> 00:18:25,640 REACTIVITY. 399 00:18:25,640 --> 00:18:27,800 MOST RECENT STYLES HAVE SHOWN 400 00:18:27,800 --> 00:18:30,160 THAT EFFECTS CONTRIBUTE TO 401 00:18:30,160 --> 00:18:33,880 INFANT HEALTH AND IN 2014 A 402 00:18:33,880 --> 00:18:36,520 NUMBER OF INFANT FORMULA 403 00:18:36,520 --> 00:18:38,120 MANUFACTURES INTRODUCED THIS AS 404 00:18:38,120 --> 00:18:40,560 AN ADDITIVE TO THEIR PRODUCT AND 405 00:18:40,560 --> 00:18:41,840 ALL THESE ACCOMPLISHMENTS OF 406 00:18:41,840 --> 00:18:45,320 COURSE WERE POSSIBLE BECAUSE OF 407 00:18:45,320 --> 00:18:45,920 THE BIOLOGICAL GLYCANS WERE 408 00:18:45,920 --> 00:18:47,120 AVAILABLE IN THE LARGE NOWOF THE 409 00:18:47,120 --> 00:18:48,400 QUANTITIES TO DO THE STUDIES. 410 00:18:48,400 --> 00:18:50,280 ANOTHER EXAMPLE OF THE FLUID 411 00:18:50,280 --> 00:18:53,080 RICH IN GLYCAN IS HUMAN OVANIAR 412 00:18:53,080 --> 00:18:54,120 ASSIST FLUID AND IN THE EARLY 413 00:18:54,120 --> 00:18:55,920 DAYS OF MEDICAL CARE, THIS WAS A 414 00:18:55,920 --> 00:18:58,000 FLUID THAT WAS READILY AVAILABLE 415 00:18:58,000 --> 00:19:00,520 AND IT CONTAINED MUSEINS THAT 416 00:19:00,520 --> 00:19:03,680 CARRIED THE ABO BLOOD GROUPS OF 417 00:19:03,680 --> 00:19:04,440 THEIR DONOR. 418 00:19:04,440 --> 00:19:05,520 AND BECAUSE OF THEIR 419 00:19:05,520 --> 00:19:06,480 AVAILABILITY AND LARGE 420 00:19:06,480 --> 00:19:11,760 QUANTITIES, THEY WERE USED AS 421 00:19:11,760 --> 00:19:13,240 THE STRUCTURES FOR DETERMINING 422 00:19:13,240 --> 00:19:16,920 THE ACTUAL SEQUENCE OF THE ABO 423 00:19:16,920 --> 00:19:18,360 AND LEWIS BLOOD WORK SYSTEMS. 424 00:19:18,360 --> 00:19:20,200 SO USING THOSE AS AN EXAMPLE, 425 00:19:20,200 --> 00:19:21,400 NDPLI KAN--KANA WAS FOUNDED ON 426 00:19:21,400 --> 00:19:23,080 THE CONCEPT THAT IF WE COULD 427 00:19:23,080 --> 00:19:25,920 MAKE MAMMALIAN DPLI KAN--KANAS 428 00:19:25,920 --> 00:19:28,960 AS AVAILABLE, AS HMOs, OR AS 429 00:19:28,960 --> 00:19:30,120 OVERYIAN SIS FLUID, WE MIGHT BE 430 00:19:30,120 --> 00:19:31,680 ABLE TO CONTRIBUTE TO ADVANCING 431 00:19:31,680 --> 00:19:33,120 OUR UNDERSTANDING OF THE DPLI 432 00:19:33,120 --> 00:19:33,960 KAN--KANA FUNCTIONS AND THIS 433 00:19:33,960 --> 00:19:40,600 SEEMS TO BE A MAJOR--A MAJOR 434 00:19:40,600 --> 00:19:42,800 GOAL OF THE SYNTHETIC EFFORTS IN 435 00:19:42,800 --> 00:19:53,600 THE COMMON FUND WORK. 436 00:19:53,600 --> 00:19:54,960 WE HAVE A NUMBER OF DIFFICULTS, 437 00:19:54,960 --> 00:19:57,120 WE KNOW THAT SYNTHESIS AND 438 00:19:57,120 --> 00:19:59,480 SEQUENCING OF LARGE AND NATURAL 439 00:19:59,480 --> 00:20:02,120 GLYCAN SYSTEM DIFFICULT, AND AS 440 00:20:02,120 --> 00:20:03,720 MENTIONED, YESTERDAY, UNLIKE 441 00:20:03,720 --> 00:20:06,720 OLIGO NUCLEOTIDES AND PROTEIN 442 00:20:06,720 --> 00:20:07,520 AMPLIFICATION OF PROTEIN 443 00:20:07,520 --> 00:20:08,760 STRUCTURES AND NOT A ROUTINE 444 00:20:08,760 --> 00:20:09,600 PROCESS, ALTHOUGH WE ARE WORKING 445 00:20:09,600 --> 00:20:14,000 ON SOME OF THOSE IDEAS. 446 00:20:14,000 --> 00:20:15,720 AND NATURAL DPLI KAN--KANAS ARE 447 00:20:15,720 --> 00:20:17,600 INDEED IN LOW ABUNDANCE IN 448 00:20:17,600 --> 00:20:18,720 NATURAL PRODUCTS. 449 00:20:18,720 --> 00:20:22,120 SO WHAT WE NEED IS A CHEMICAL 450 00:20:22,120 --> 00:20:26,720 METHOD THAT WE CAN SCALE UP SO 451 00:20:26,720 --> 00:20:28,680 AT NET TPHREU RACING KAN--KANA 452 00:20:28,680 --> 00:20:30,920 WE START WITH LARGE QUANTITIES 453 00:20:30,920 --> 00:20:33,360 OF NATURAL PRODUCTS AND THEN WE 454 00:20:33,360 --> 00:20:34,760 APPLY ORANGE AND SOANCH A 455 00:20:34,760 --> 00:20:36,520 SCALABLE PROCESS AND WE 456 00:20:36,520 --> 00:20:39,320 DETERMINED AT THIS POINT, THAT 457 00:20:39,320 --> 00:20:44,120 THE SCALE OF OF THE PROCESS IS 458 00:20:44,120 --> 00:20:45,440 LARGELY DEPENDENTOT SIZE OF THE 459 00:20:45,440 --> 00:20:47,680 EQUIPMENT AND IF WE CAN 460 00:20:47,680 --> 00:20:49,360 CAPITALIZE ON THIS ECONOMY OF 461 00:20:49,360 --> 00:20:50,240 SCALE AND WE'RE WORKING ON THAT, 462 00:20:50,240 --> 00:20:53,000 IT MAY BE POSSIBLE REALLY TO 463 00:20:53,000 --> 00:20:54,320 SUPPLY MILLIGRAM QUANTITIES OF 464 00:20:54,320 --> 00:21:00,720 GLYCANS AT VERY LOW COSTS. 465 00:21:00,720 --> 00:21:02,720 NOW THANKS TO NIGMS TO DR. SONG 466 00:21:02,720 --> 00:21:06,520 IN THE EARLY DAYS AND PHASE 1 467 00:21:06,520 --> 00:21:08,640 PROJECT, TO NET GLYCAN, WE 468 00:21:08,640 --> 00:21:10,880 DEMONSTRATED THE FEASIBLE OF 469 00:21:10,880 --> 00:21:11,320 THIS APPROACH. 470 00:21:11,320 --> 00:21:13,120 AND AS I'M SHOWING YOU HERE, YOU 471 00:21:13,120 --> 00:21:14,800 SEE THE SLIDE BEFORE, BUT THIS 472 00:21:14,800 --> 00:21:17,720 JUST SHOWS THE ORANGE PROCESS AS 473 00:21:17,720 --> 00:21:19,400 IT--CAN REMOVE DPLI KAN--KANAS 474 00:21:19,400 --> 00:21:22,960 FROM ANY GLYCO CONJUGATE, BUT WE 475 00:21:22,960 --> 00:21:30,120 FOCUSED ONURE TECH ON NGLYCANS. 476 00:21:30,120 --> 00:21:31,200 NOW AFTER DEMONSTRATING THESE 477 00:21:31,200 --> 00:21:32,720 ABILITY, WE WERE AWARDED A PHASE 478 00:21:32,720 --> 00:21:36,880 2 GRANT WHICH IS NOW SUPPORTING 479 00:21:36,880 --> 00:21:38,120 THE NRM PRODUCTION OF DPLRKS 480 00:21:38,120 --> 00:21:39,480 DPLI KAN--KANAS AND WE 481 00:21:39,480 --> 00:21:41,880 ACCOMPLISHED THIS BY 482 00:21:41,880 --> 00:21:45,440 THEOXIDATING PROCESS TO CONTROL 483 00:21:45,440 --> 00:21:47,040 BYPRODUCTS BY PREPARING BI 484 00:21:47,040 --> 00:21:48,320 FUNCTIONAL TAGS SO YOU FOLLOW 485 00:21:48,320 --> 00:21:50,320 THE PUREUNCTION AND THEN THESE 486 00:21:50,320 --> 00:21:52,120 CAN BE IMMOBILIZED FOR A VARIETY 487 00:21:52,120 --> 00:21:54,440 OF FUNCTIONAL STUDIES AND WE'VE 488 00:21:54,440 --> 00:21:55,800 DEVELOPED REMOVABLE TAGS TO 489 00:21:55,800 --> 00:21:57,520 FOLLOW PURIFICATION AND THEN, 490 00:21:57,520 --> 00:21:59,080 REMOVE THEM SO THAT WE CAN HAVE 491 00:21:59,080 --> 00:22:02,600 ACCESS TO THE FREE NATURAL 492 00:22:02,600 --> 00:22:04,280 GLYCANS. 493 00:22:04,280 --> 00:22:10,880 AND WE'VE DEVELOPED 494 00:22:10,880 --> 00:22:11,480 PREVENTIVERATIVE PURIFICATION 495 00:22:11,480 --> 00:22:13,200 METHODS TO PURIFY DPLI 496 00:22:13,200 --> 00:22:14,520 KAN--KANAS, AND THIS 497 00:22:14,520 --> 00:22:15,240 PURIFICATION PROCESS AS EVERYONE 498 00:22:15,240 --> 00:22:18,080 TOLD US EARLY ON WAS THE IMPABL 499 00:22:18,080 --> 00:22:20,640 PART IT WAS DIFFICULT AND WE'RE 500 00:22:20,640 --> 00:22:26,520 STILL WORKING ON IT BUT IT IS 501 00:22:26,520 --> 00:22:29,280 PROGRESSING AS I SAID WE FOCUSED 502 00:22:29,280 --> 00:22:30,280 OUR FIRST ACTIVITYOT HIGH 503 00:22:30,280 --> 00:22:31,280 MANNERS IN DPLI KAN--KANAS AND 504 00:22:31,280 --> 00:22:33,640 WE'RE HOW HAPPY TO REPORT THAT 505 00:22:33,640 --> 00:22:36,760 WE HAVE OVER A GRAM OF MAN 9 IN 506 00:22:36,760 --> 00:22:38,720 INVENTORY AND A HUNDRED 507 00:22:38,720 --> 00:22:40,520 MILLIGRAM QUANTITIES OF THE 508 00:22:40,520 --> 00:22:42,720 MAJOR ISOMERS OF THE STRUCTURES, 509 00:22:42,720 --> 00:22:43,760 NOW THE MINORS THERE, ARE 510 00:22:43,760 --> 00:22:45,120 CERTAINLY THERE BUT IN SMALLER 511 00:22:45,120 --> 00:22:48,640 QUANTITIES BUT THEY ARE THERE 512 00:22:48,640 --> 00:22:49,360 THESE DPLI KAN--KANAS ARE 513 00:22:49,360 --> 00:22:50,960 AVAILABLE IN A VARIETY OF 514 00:22:50,960 --> 00:22:56,240 DERIVATIVES THAT CAN BE USED FOR 515 00:22:56,240 --> 00:22:57,720 STANDARDS AND MARKERS WE HAVE 516 00:22:57,720 --> 00:22:59,400 THEM WITH FUNCTIONAL GROUPS THAT 517 00:22:59,400 --> 00:23:00,960 CAN BE ATTACHED TO VARIOUS 518 00:23:00,960 --> 00:23:02,120 IMMOBILIZED CERTAINTIES AND WE 519 00:23:02,120 --> 00:23:04,760 CAN MAKE THEM WITH BIOTIN AS 520 00:23:04,760 --> 00:23:08,560 WELL SO THESE ARE READILY 521 00:23:08,560 --> 00:23:11,920 AVAILABLE FOR ANALYSIS, SO THIS 522 00:23:11,920 --> 00:23:13,600 MEANS THAT WE CAN NOW SAY THAT 523 00:23:13,600 --> 00:23:15,760 WE HAVE LARGE QUANTITIES OF PURE 524 00:23:15,760 --> 00:23:17,320 NATURAL GLYCANS AVAILABLE AS 525 00:23:17,320 --> 00:23:20,120 NEEDED TO DEFINE GLYCAN 526 00:23:20,120 --> 00:23:20,400 STRUCTURE. 527 00:23:20,400 --> 00:23:21,800 NOW THE CHALLENGE TO OUR 528 00:23:21,800 --> 00:23:22,960 COMMUNITY, I BELIEVE NOW IS WHAT 529 00:23:22,960 --> 00:23:25,960 DO WE DO WITH THESE OLIGO 530 00:23:25,960 --> 00:23:26,280 SACCHARIDES. 531 00:23:26,280 --> 00:23:28,080 OUR HOPE IS THAT FOLKS WILL WANT 532 00:23:28,080 --> 00:23:30,560 TO PURCHASE THEM AND USE THEM IN 533 00:23:30,560 --> 00:23:32,240 THE EXPERIMENTS FOR GLUE MARIOUS 534 00:23:32,240 --> 00:23:34,160 KOAMIC STUDIES AND SOME OF THOSE 535 00:23:34,160 --> 00:23:35,920 STUDIES MIGHT INCLUDE THINGS 536 00:23:35,920 --> 00:23:39,120 I'VE LISTED HERE, WE NOW REALLY 537 00:23:39,120 --> 00:23:40,360 HAVE UNLIMITED QUANTITIES THAT 538 00:23:40,360 --> 00:23:42,320 CAN BE USED FOR STAND ARPDS FOR 539 00:23:42,320 --> 00:23:44,400 METHODS OF DEVELOPMENT AND 540 00:23:44,400 --> 00:23:45,560 DEFINING STRUCTURES. 541 00:23:45,560 --> 00:23:48,240 SINCE WE HAVE--WE CAN MAKE 542 00:23:48,240 --> 00:23:50,880 LARGER AMOUNTS, WE CAN USE THESE 543 00:23:50,880 --> 00:23:52,440 AS PRECURSOR FOR ELONGATION OF 544 00:23:52,440 --> 00:23:55,400 OTHER STRUCTURE BY ENZYMATIC 545 00:23:55,400 --> 00:23:59,520 METHODS, AS DR. SONG MENTIONED 546 00:23:59,520 --> 00:24:02,600 THE SUBSTRATES FOR THE MUSEUM 547 00:24:02,600 --> 00:24:04,560 STATED END-TO-END IS A MAJOR 548 00:24:04,560 --> 00:24:07,320 BYPRODUCT IS A MAJOR DERIVATIVE 549 00:24:07,320 --> 00:24:08,640 AND DR. LEE'S GOING TO TALK 550 00:24:08,640 --> 00:24:25,280 ABOUT THAT IN A LITTLE WHILE. 551 00:24:25,280 --> 00:24:26,000 CO CRYSTALLIZE [INDISCERNIBLE] 552 00:24:26,000 --> 00:24:27,960 CAN BE USED FOR ISOLATION FOR 553 00:24:27,960 --> 00:24:30,280 PULL DOWN EXPERIMENTS TO PULL 554 00:24:30,280 --> 00:24:33,760 DOWN SPECIFIC GLYCAN BIND 555 00:24:33,760 --> 00:24:34,800 PROGTEENS, GLYCAN CONJUGATES CAN 556 00:24:34,800 --> 00:24:37,520 BE USED TO SCREEN FOR ANTIGLYCAN 557 00:24:37,520 --> 00:24:38,880 REAGENTS BY PHASIA'S PLAY AND 558 00:24:38,880 --> 00:24:40,360 OTHER EFFORTS AND DR. CUMMINGS 559 00:24:40,360 --> 00:24:43,040 HAS DONE A LOT OF WORK IN THAT 560 00:24:43,040 --> 00:24:45,080 AREA NADDITION TO CELLULAR 561 00:24:45,080 --> 00:24:46,520 RESPONSES THAT DEFINE FUNCTION 562 00:24:46,520 --> 00:24:47,920 FUNCTIONS OF POSSIBILITY AND 563 00:24:47,920 --> 00:24:49,840 EVEN ANIMAL STUDIES ON SMALL 564 00:24:49,840 --> 00:24:52,960 ANIMALS FOR LOOKING AT POTENTIAL 565 00:24:52,960 --> 00:24:56,080 SYSTEMIC EFFECTS OF NGLYCANS OR 566 00:24:56,080 --> 00:24:56,760 OTHERWISE ADMINISTERING DPLI 567 00:24:56,760 --> 00:24:57,960 KAN--KANAS CAN BE POSSIBLE FOR 568 00:24:57,960 --> 00:25:01,000 WE'RE LOOKING FOR PEOPLE TO 569 00:25:01,000 --> 00:25:02,560 BEGIN APPROACHING AND THINKING 570 00:25:02,560 --> 00:25:03,880 OUTSIDE THE BOX ABOUT OTHER 571 00:25:03,880 --> 00:25:04,920 METHODS THAT CAN BE USED TO DO 572 00:25:04,920 --> 00:25:06,720 THIS AND WE HAVE A NUMBER OF 573 00:25:06,720 --> 00:25:09,080 GLYCANS NOW, IN OUR INVENTORY. 574 00:25:09,080 --> 00:25:10,040 IRWANT TO BREFLY GO THROUGH 575 00:25:10,040 --> 00:25:12,240 THESE, THESE ARE THE COMPLEX 576 00:25:12,240 --> 00:25:13,280 BINARY TPHREU RACING KAN--KANAS 577 00:25:13,280 --> 00:25:20,520 WE HAVE THEM IN PRIMARILY IN THE 578 00:25:20,520 --> 00:25:21,120 SIXTH SIGNIFIAALATED 579 00:25:21,120 --> 00:25:23,480 [INDISCERNIBLE]. 580 00:25:23,480 --> 00:25:25,480 THESE ARE HUNDRED MILLIGRAM 581 00:25:25,480 --> 00:25:25,760 QUANTITIES. 582 00:25:25,760 --> 00:25:27,720 WE HAVE THE DERIVATIVES OF ALL 583 00:25:27,720 --> 00:25:29,480 OF THESE STRUCTURES. 584 00:25:29,480 --> 00:25:31,600 WE'VE ISOLATED A NUMBER OF 585 00:25:31,600 --> 00:25:32,000 HYBRID STRUCTURES. 586 00:25:32,000 --> 00:25:33,320 THESE ARE SOME OF THE MAJOR 1S 587 00:25:33,320 --> 00:25:35,000 WE HAVE, THERE ARE A LOT MORE 588 00:25:35,000 --> 00:25:37,280 COMES ALONG, AND IT'S COMING 589 00:25:37,280 --> 00:25:39,720 SOON, ALSO WILL BE THE 590 00:25:39,720 --> 00:25:40,720 MULTIITENIARY GLYCANS THAT ARE 591 00:25:40,720 --> 00:25:41,720 WORKING ON THOSE. 592 00:25:41,720 --> 00:25:44,120 SO IN SUMMARY, I WANT TO JUST 593 00:25:44,120 --> 00:25:46,040 SAY THAT NET DPLI KAN--KANA 594 00:25:46,040 --> 00:25:48,120 SPECIALIZING IN NATURAL 595 00:25:48,120 --> 00:25:51,320 GIVING--YOU KAN--KANAS FOR 596 00:25:51,320 --> 00:25:52,280 FUNCTIONAL GLYCANS. 597 00:25:52,280 --> 00:25:55,120 WE HAVE NDPLI KAN--KANAS AND 598 00:25:55,120 --> 00:25:57,720 NGRAM QUANTITIES WITH PURITYS 599 00:25:57,720 --> 00:25:59,880 GREATER THAN 90%. 600 00:25:59,880 --> 00:26:01,160 OGLYCANS AND NGLYCANS ARE COMING 601 00:26:01,160 --> 00:26:02,520 IN THE FUTURE AND WE'RE PREPARED 602 00:26:02,520 --> 00:26:04,160 TO WORK WITH ANYBODY WHO WANTS 603 00:26:04,160 --> 00:26:05,720 TO FIND THE DPLI KAN--KANA OUT 604 00:26:05,720 --> 00:26:05,920 THERE. 605 00:26:05,920 --> 00:26:13,800 WE FEEL LIKE THEY'RE THERE, AND 606 00:26:13,800 --> 00:26:16,160 AND WE SHOULD BE ABLE TO 607 00:26:16,160 --> 00:26:18,280 IDENTIFY THEM FOR YOU AND WE 608 00:26:18,280 --> 00:26:21,400 FEEL LIKE THIS IS IN LINE WITH 609 00:26:21,400 --> 00:26:21,760 FEDERAL GRANTS. 610 00:26:21,760 --> 00:26:23,000 WITH THAT I WANT TO SAY THANK 611 00:26:23,000 --> 00:26:24,240 YOU FOR YOUR TIME AND WE LOOK 612 00:26:24,240 --> 00:26:26,440 FORWARD TO YOUR ORDERS. 613 00:26:26,440 --> 00:26:27,120 [ APPLAUSE ] 614 00:26:27,120 --> 00:26:27,520 EMPLOY. 615 00:26:27,520 --> 00:26:28,720 >> THANK YOU SO MUCH DR. SMITH. 616 00:26:28,720 --> 00:26:34,000 AND CONTINUING IN OUR THEME OF 617 00:26:34,000 --> 00:26:35,040 BASIC SCIENCE TRANSLATED TO 618 00:26:35,040 --> 00:26:39,720 COMPANIES OR OTHER BASIC BIOLOGY 619 00:26:39,720 --> 00:26:40,000 STUDIES, I INTRODUCED 620 00:26:40,000 --> 00:27:14,000 DR. PRUDDEN AND BOONES. 621 00:27:14,000 --> 00:27:15,520 --SERIES OF DPLI COSDACE AND 622 00:27:15,520 --> 00:27:18,480 TRANSFER ACES TO CONSTRUCT A 623 00:27:18,480 --> 00:27:21,480 NUMBER OF IMPORTANT SUBCLASSES 624 00:27:21,480 --> 00:27:23,320 OF NGLYCANS BEING 625 00:27:23,320 --> 00:27:25,840 [INDISCERNIBLE] STRUCTURES 626 00:27:25,840 --> 00:27:27,200 COMPLEX AND HYBRID STRUCTURES. 627 00:27:27,200 --> 00:27:38,080 THE TALK TODAY WILL FOCUS ON THE 628 00:27:38,080 --> 00:27:38,640 COMPLEX AMONG THEMSELVES. 629 00:27:38,640 --> 00:27:39,720 SO THERE ARE A NUMBER OF 630 00:27:39,720 --> 00:27:40,840 IMPORTANCES THAT HAVE BEEN 631 00:27:40,840 --> 00:27:42,120 OBSERVED WITH NDPLI KAN--KANAS 1 632 00:27:42,120 --> 00:27:43,440 OF THEM BEING THE MOST IT IS 1 633 00:27:43,440 --> 00:27:47,640 OF THE MOST COMMON AND COMPLEX 634 00:27:47,640 --> 00:27:48,120 POST TRANSLATIONAL 635 00:27:48,120 --> 00:27:48,440 MODIFICATIONS. 636 00:27:48,440 --> 00:27:52,200 THEY'RE RELEVANT TO A NUMBER OF 637 00:27:52,200 --> 00:27:53,440 BIOLOGICAL PROCESSES, AND 638 00:27:53,440 --> 00:27:55,280 GLYCOSYLATION OF ANTIBODIES HAS 639 00:27:55,280 --> 00:27:57,240 BEEN SHOWN TO MEDIATE CELL 640 00:27:57,240 --> 00:27:57,920 CYTOTOXICITY. 641 00:27:57,920 --> 00:28:00,320 WE ALSO KNOW THAT CHANGES IN 642 00:28:00,320 --> 00:28:02,320 DPLI COSALATION OF PROFILE CAN 643 00:28:02,320 --> 00:28:03,640 CHANGE DURING DISEASE STATES 644 00:28:03,640 --> 00:28:06,040 WHICH MAKES DPLI COSALATION AN 645 00:28:06,040 --> 00:28:08,480 INTERESTING TARGET FOR BIOMARKER 646 00:28:08,480 --> 00:28:08,840 DEVELOPMENTS. 647 00:28:08,840 --> 00:28:09,640 ALTHOUGH THE IMPORTANCES ARE 648 00:28:09,640 --> 00:28:11,200 WELL UNDERSTOOD, THERE ARE A 649 00:28:11,200 --> 00:28:12,680 NUMBER OF CHALLENGES, SOME OF 650 00:28:12,680 --> 00:28:16,520 WHICH WE'VE ALREADY HEARD, 651 00:28:16,520 --> 00:28:17,800 HETEROGENERATED AITY, OF N-DPLI 652 00:28:17,800 --> 00:28:19,640 KAN--KANA CANS DEPEND ON CELL 653 00:28:19,640 --> 00:28:22,080 TYPE AND MICRO ENVIRONMENT. 654 00:28:22,080 --> 00:28:23,200 THESE STRUCTURES ARE ISOBERNEA 655 00:28:23,200 --> 00:28:25,680 DEETIC, MEANING THAT THEY CAN 656 00:28:25,680 --> 00:28:29,720 HAVE THE SAME MONOSACCHARIDE 657 00:28:29,720 --> 00:28:31,120 CONNECTIVITY, OR MONOSACCHARIDE 658 00:28:31,120 --> 00:28:33,440 PROFILE, WITH DIFFERENT 659 00:28:33,440 --> 00:28:34,720 CONNECTIVITY GIVING THEM 660 00:28:34,720 --> 00:28:35,320 IDENTICAL MOLECULAR WEIGHTS. 661 00:28:35,320 --> 00:28:37,440 IT CAN BE ALTHOUGH THERE HAVE 662 00:28:37,440 --> 00:28:38,840 BEEN IMPROVEMENTS DIFFICULTY IN 663 00:28:38,840 --> 00:28:39,920 ASSESSING INDIVIDUAL WELL 664 00:28:39,920 --> 00:28:52,840 DEFINED STRUCTURES FROM NATURE 665 00:28:52,840 --> 00:28:57,640 AND SUCH ISOLATION DEPENDS ON 666 00:28:57,640 --> 00:28:59,240 [INDISCERNIBLE], ADDITIONAL 667 00:28:59,240 --> 00:29:01,400 SOURCES ARE COMING ON ONLINE AND 668 00:29:01,400 --> 00:29:05,680 THE GOAL OF OUR NIH SBIR IS TO 669 00:29:05,680 --> 00:29:07,200 UTILIZE THE ENZYMATIC 670 00:29:07,200 --> 00:29:07,960 METHODOLOGY FOR SIPGHTICIZING 671 00:29:07,960 --> 00:29:09,760 THE 20 MOST ABUPPED ANT GLYCANS 672 00:29:09,760 --> 00:29:11,200 DONE IN HUMAN SERUM, ALL OF 673 00:29:11,200 --> 00:29:14,040 WHICH HAVE A GREAT--HAVE AN 674 00:29:14,040 --> 00:29:15,040 ABUNDANCE GREATER THAN 1%, 675 00:29:15,040 --> 00:29:18,640 TOGETHER THAT MAKES UP 50% FOR 676 00:29:18,640 --> 00:29:21,520 THE TOTAL SERUM NDPLI KOAM, 677 00:29:21,520 --> 00:29:22,640 UNFORTUNATELY A SIGNIFICANT 678 00:29:22,640 --> 00:29:24,480 POPULATION OF PROTEINS WHICH 679 00:29:24,480 --> 00:29:26,120 INCLUDE PROTEINS THAT ARE 680 00:29:26,120 --> 00:29:28,080 GLYCOSYLATED, THE LIBRARY WE ARE 681 00:29:28,080 --> 00:29:28,920 60ICIZEING AS INDIVIDUAL 682 00:29:28,920 --> 00:29:33,680 STRUCTURES WILL CONTAIN A FREE 683 00:29:33,680 --> 00:29:34,840 REDUCING TERMINUS, REDUCTIVE 684 00:29:34,840 --> 00:29:36,440 AMDISMAGZ THIS ALLOWS FOR 685 00:29:36,440 --> 00:29:39,760 EXTREME FLEXIBILITY AND USER 686 00:29:39,760 --> 00:29:44,840 WHETHER IT'S LCMS, MSMSOR OR CAP 687 00:29:44,840 --> 00:29:46,040 ILLEGALSARY ELECTROFORRIESIES, 688 00:29:46,040 --> 00:29:49,280 THE PRODUCT IS AMENABLE TO SUCH 689 00:29:49,280 --> 00:29:50,640 APPLICATIONS, ADDITIONALLY THESE 690 00:29:50,640 --> 00:29:54,240 ARE PURIFIED BY LCMS AND THEIR 691 00:29:54,240 --> 00:29:55,800 STRUCTURES ARE CONFIRMED BY NMR, 692 00:29:55,800 --> 00:29:58,760 AND MOST IMPORTANTLY THE LIBRARY 693 00:29:58,760 --> 00:30:02,600 TO BE SYNTHESIZED WILL CONTAIN A 694 00:30:02,600 --> 00:30:05,440 UNIFORMLY CLICK NAC WHICH CAN BE 695 00:30:05,440 --> 00:30:08,160 USED ANAIN ERNE TERNAL 696 00:30:08,160 --> 00:30:08,600 [INDISCERNIBLE]. 697 00:30:08,600 --> 00:30:10,840 MAKING THIS PRODUCT CAPABLE OF 698 00:30:10,840 --> 00:30:12,280 DETERMINING DPLI KAN--KANA 699 00:30:12,280 --> 00:30:14,880 CONCENTRATION CHANGES WITH 700 00:30:14,880 --> 00:30:16,160 REGARDS TO DISEASE PROGRESSIONS, 701 00:30:16,160 --> 00:30:21,000 THIS WILL BE IMPORTANT FOR THE 702 00:30:21,000 --> 00:30:21,400 BIOMARKER SECTOR. 703 00:30:21,400 --> 00:30:24,760 AS YOU CAN SEE, THEY ARE VERY 704 00:30:24,760 --> 00:30:27,080 DIVERSE, THERE ARE 705 00:30:27,080 --> 00:30:28,240 NATURAL--NEUTRAL STRUCTURES, WE 706 00:30:28,240 --> 00:30:30,800 HAVE A, SIDIC STRUCTURES BEARING 707 00:30:30,800 --> 00:30:39,160 1 OR 2 SALIC ACIDS, THESE 708 00:30:39,160 --> 00:30:41,040 SIALATED CAN BE ASYMMETRIC WHERE 709 00:30:41,040 --> 00:30:45,240 WE HAVE DIFFERENT 1S IN EACH 710 00:30:45,240 --> 00:30:46,560 ARM, [INDISCERNIBLE] GLUE 711 00:30:46,560 --> 00:30:52,240 COSALATED AND ALSO STRUCTURES 712 00:30:52,240 --> 00:30:53,760 WITH ISOMERIC SIALICK ACID 713 00:30:53,760 --> 00:30:55,440 PLACEMENT ALTHOUGH WE HAVE SENT 714 00:30:55,440 --> 00:30:56,680 OUT THE TARGETS, THE PROTOCOL WE 715 00:30:56,680 --> 00:30:59,480 USE IN THE NEXT SLIDE OR 2, CAN 716 00:30:59,480 --> 00:31:02,320 PROVIDE ACCESS TO OTHER ADVANCED 717 00:31:02,320 --> 00:31:04,880 INTERMEDIATE THAT WILL BE USEFUL 718 00:31:04,880 --> 00:31:06,840 STANDARDS, SO OUR STRATEGY 719 00:31:06,840 --> 00:31:09,720 RELIES ON AN ADVANCED STARTING 720 00:31:09,720 --> 00:31:11,640 MATERIAL THAT WE EXTRACT FROM 721 00:31:11,640 --> 00:31:15,000 THE EGG YOLK POWDER, THAT 722 00:31:15,000 --> 00:31:16,960 EXTRACTED MATERIAL IS DIGESTED 723 00:31:16,960 --> 00:31:18,920 TO FOR THE [INDISCERNIBLE] 724 00:31:18,920 --> 00:31:20,120 SACCHARIDE WHICH IS COMMON TO 725 00:31:20,120 --> 00:31:22,120 ALL NDPLI KAN--KANAS. 726 00:31:22,120 --> 00:31:25,680 OR ALL HUMAN NGLAI KAN--KANAS. 727 00:31:25,680 --> 00:31:29,120 AND VERY BRIEFLY, THIS IS HOW WE 728 00:31:29,120 --> 00:31:32,840 INSTALL OUR C13 LABELED TAG. 729 00:31:32,840 --> 00:31:38,880 STARTING FROM THE CORE 730 00:31:38,880 --> 00:31:39,880 APENTOSACCHARIDE, AND USE GLYNAC 731 00:31:39,880 --> 00:31:43,440 ON INSTALL THE ARM TO CREATE THE 732 00:31:43,440 --> 00:31:46,240 DESIRED HEXASACCHARIDE. 733 00:31:46,240 --> 00:31:48,400 THIS IS HEXASACCHARIDE IS FOR 734 00:31:48,400 --> 00:31:51,120 MDN2, AND WE CAN USE THE CARBON 735 00:31:51,120 --> 00:31:55,720 13 TO SELECT THE INSTALL 1, C13 736 00:31:55,720 --> 00:31:57,840 LABELED GLYCNAC ON THE BRANCH 737 00:31:57,840 --> 00:31:59,240 TREATING THE SACCHARIDE. 738 00:31:59,240 --> 00:32:04,240 THIS HELPED THE SACCHARIDE THEN 739 00:32:04,240 --> 00:32:06,360 CAN BE EXTENDED IT COULD BE 740 00:32:06,360 --> 00:32:09,840 EXTENDED USING TRANSFER ACES, TO 741 00:32:09,840 --> 00:32:13,320 CREATE 2 LAC INCOME AY, AND THEN 742 00:32:13,320 --> 00:32:16,960 DOZE CAN BE MODIFIED WITH 743 00:32:16,960 --> 00:32:18,680 VARIOUS SILO TRANSFERAISES TO 744 00:32:18,680 --> 00:32:20,680 CREATE THE DESIRED TARGETS. 745 00:32:20,680 --> 00:32:22,040 IMPORTANTLY AS CAN YOU SEE FROM 746 00:32:22,040 --> 00:32:25,920 THE SCREAM ISSUES NOT ONLY DO WE 747 00:32:25,920 --> 00:32:28,560 GET THE DESIRED DISCIPLINARY 748 00:32:28,560 --> 00:32:30,800 SOLID STRUCTURES, BUT ALSO 749 00:32:30,800 --> 00:32:34,240 INTERMEDIATE PROVIDE TARGETS OF 750 00:32:34,240 --> 00:32:35,240 THEIR OWN IMPORTANCE. 751 00:32:35,240 --> 00:32:39,120 FURTHER MORE THE INSTALLATION OF 752 00:32:39,120 --> 00:32:42,360 A UNIFORMLY INSTALLED GLYCNAC AT 753 00:32:42,360 --> 00:32:45,440 A PARTICULAR SITE DOES ALLOW FOR 754 00:32:45,440 --> 00:32:46,240 FRAG M-TEBURKEULOSEISATION 755 00:32:46,240 --> 00:32:47,080 ANALYSIS WHICH IS MORE DIFFICULT 756 00:32:47,080 --> 00:32:54,000 ON A STRUCTURE THAT IS NOT C13 757 00:32:54,000 --> 00:32:54,360 LABELED. 758 00:32:54,360 --> 00:32:55,680 AS I MENTIONED IN THE PAST, WE 759 00:32:55,680 --> 00:32:58,240 LOOK TO MAKE THESE PURE AND WE 760 00:32:58,240 --> 00:33:00,640 DO THAT BY USING CHROMOING TO 761 00:33:00,640 --> 00:33:01,720 RAFALLIC TECHNIQUES, I 762 00:33:01,720 --> 00:33:02,880 ILLUSTRATED 1 EXAMPLE HERE WHERE 763 00:33:02,880 --> 00:33:05,680 WE CAN INSTALL THE CORE FUCOS, 764 00:33:05,680 --> 00:33:07,600 AND USING HELIOS POSITIVIC LCMS, 765 00:33:07,600 --> 00:33:13,360 WE WERE ABLE TO SUG95 CAN'TLY 766 00:33:13,360 --> 00:33:15,200 DIFFERENTIATE OUR DESIRED 767 00:33:15,200 --> 00:33:19,640 PRODUCT FROM ANY UNREACTED SUGAR 768 00:33:19,640 --> 00:33:21,200 NUCLEOTIDES FOR REACTION 769 00:33:21,200 --> 00:33:21,480 COMPONENTS. 770 00:33:21,480 --> 00:33:25,440 THIS HOW WE'RE ABLE TO GET HIGH 771 00:33:25,440 --> 00:33:27,160 PURITY STRUCTURES, WITH ONLY 1 772 00:33:27,160 --> 00:33:29,240 TARGET SO THERE WILL BE NO 773 00:33:29,240 --> 00:33:40,640 MIXTURES IN THESE PRODUCTS. 774 00:33:40,640 --> 00:33:43,240 WE ARE LOOKING AT A VAST LIBRARY 775 00:33:43,240 --> 00:33:45,480 TO LOOK AT STANDARDS ISSUES THE 776 00:33:45,480 --> 00:33:46,960 STARTING MATERIAL THAT WE BEBIN 777 00:33:46,960 --> 00:33:50,120 WITH IS EXTRACTED FROM EGG YOLK 778 00:33:50,120 --> 00:33:53,880 POWDER AND WE USE RECOMBIN ANT 779 00:33:53,880 --> 00:33:56,280 STRANS FER ACES TO YIELD 780 00:33:56,280 --> 00:34:02,240 ISOMERS, WE USE AN UNNATURAL 781 00:34:02,240 --> 00:34:03,000 GLYCNAC TO ASYMMETRIC TARGETS 782 00:34:03,000 --> 00:34:04,200 WHICH I DON'T HAVE TIME TO 783 00:34:04,200 --> 00:34:05,560 DISCUSS HERE BUT THIS IS 784 00:34:05,560 --> 00:34:09,560 AVAILABLE ON A PAPER WE PUBLISH 785 00:34:09,560 --> 00:34:11,120 INDEED 2019. 786 00:34:11,120 --> 00:34:13,760 HPLC PURIFICATION RESULTS IN 787 00:34:13,760 --> 00:34:14,360 ANALYTICALLY PURE MATERIAL, I 788 00:34:14,360 --> 00:34:19,520 WANT TO STRESS THESE HAVE FREE 789 00:34:19,520 --> 00:34:20,400 REDUCING--PREE REDUCING TARGETS 790 00:34:20,400 --> 00:34:24,440 SO THEY'RE AMENABLE TO A NUMBER 791 00:34:24,440 --> 00:34:24,840 OF APPLICATIONS. 792 00:34:24,840 --> 00:34:27,800 COMPREHENSIVE INCLUSION OF 793 00:34:27,800 --> 00:34:28,960 UNIFORMLY LABELED CARBON 13 794 00:34:28,960 --> 00:34:30,560 REPRESENTS A UNIQUE COLLECTION 795 00:34:30,560 --> 00:34:32,320 OF TARGETS THAT COMPRISE WITH 796 00:34:32,320 --> 00:34:34,640 50% OF THE SERUM DPLI KOAM AND 797 00:34:34,640 --> 00:34:38,360 WE CURRENTLY ARE IN DISCUSSIONS, 798 00:34:38,360 --> 00:34:39,520 COMMERCIAL PARTNERING WITH 799 00:34:39,520 --> 00:34:42,320 DISCUSSIONS AS THEY HAVE AN 800 00:34:42,320 --> 00:34:44,160 INTEREST IN WORKING WITH US ON 801 00:34:44,160 --> 00:34:49,440 THESE CARBON 13 LABELED TARGETS. 802 00:34:49,440 --> 00:34:51,840 SO WITH THAT JUST A COUPLE OF 803 00:34:51,840 --> 00:34:53,720 ACKNOWLEDGMENTS OF COURSE THIS 804 00:34:53,720 --> 00:34:56,040 FUNDING WAS PROVIDED BY THE NIH 805 00:34:56,040 --> 00:35:00,160 THROUGH A CURRENT SBIR PHASE 1 806 00:35:00,160 --> 00:35:02,040 AND THIS--THIS TECHNOLOGY WAS 807 00:35:02,040 --> 00:35:03,520 ACTUALLY STARTED DURING MY 808 00:35:03,520 --> 00:35:07,280 Ph.D. WORK WHICH WAS FUNDED BY 809 00:35:07,280 --> 00:35:09,240 A PREDOCTORAL FELLOWSHIP SO I 810 00:35:09,240 --> 00:35:11,280 HAVE TO GIVE ANOTHER HATS OFF TO 811 00:35:11,280 --> 00:35:14,000 NIH AS WELL, OF COURSE THE 812 00:35:14,000 --> 00:35:15,200 COMPLEX CARBOHYDRATE CENTER AND 813 00:35:15,200 --> 00:35:20,280 WE HAVE 2 WEB SITES THAT ARE 814 00:35:20,280 --> 00:35:20,840 UNDER CONSTRUCTION. 815 00:35:20,840 --> 00:35:26,240 THIS WORK IS BEING DONE OF 816 00:35:26,240 --> 00:35:28,560 COURSE ALSO WITH 817 00:35:28,560 --> 00:35:30,280 DR. [INDISCERNIBLE], AND 818 00:35:30,280 --> 00:35:31,040 DR. [INDISCERNIBLE]. 819 00:35:31,040 --> 00:35:33,160 IF ANYONE HAS ANY FURTHER 820 00:35:33,160 --> 00:35:34,160 QUESTIONS CONTACT US WITH E-MAIL 821 00:35:34,160 --> 00:35:36,640 AND I APPRECIATE THE TIME TO 822 00:35:36,640 --> 00:35:40,240 SPEAK TODAY. 823 00:35:40,240 --> 00:35:40,640 >> MANY THANKS. 824 00:35:40,640 --> 00:35:41,000 [ APPLAUSE ] 825 00:35:41,000 --> 00:35:45,280 OUR NEXT SPEAKER IS LIVE WITH US 826 00:35:45,280 --> 00:36:01,400 HERE IN BETHESDA, DR. LI FROM 827 00:36:01,400 --> 00:36:01,920 GEORGIA STATE UNIVERSITY. 828 00:36:01,920 --> 00:36:04,480 >> FIRST OF ALL I WOULD LIKE TO 829 00:36:04,480 --> 00:36:09,840 EXTEND COLLEAGUES AT THE NE H 830 00:36:09,840 --> 00:36:12,000 ORGANIZE THIS WONDERFUL MEETING 831 00:36:12,000 --> 00:36:13,320 AND IT'S REALLY A GREAT 832 00:36:13,320 --> 00:36:14,800 ALTERNATIVE FOR US TO MEET 833 00:36:14,800 --> 00:36:17,880 TOGETHER, ESPLEE FOR ME AS A 834 00:36:17,880 --> 00:36:19,320 JUNIOR FACULTY TO MEET ALL THE 835 00:36:19,320 --> 00:36:27,000 OTHERS TO TALK WITH THEM, TO I 836 00:36:27,000 --> 00:37:00,800 WILL TALK AND OGLYCANS, I HAVE A 837 00:37:00,800 --> 00:37:01,040 POINTER. 838 00:37:01,040 --> 00:37:03,360 SO I WILL START WITH THIS 839 00:37:03,360 --> 00:37:07,080 [INDISCERNIBLE] I WANT TO SHOW 840 00:37:07,080 --> 00:37:17,200 THIS BEFORE, SO, POINTER--OH, 841 00:37:17,200 --> 00:37:19,040 OKAY GO, BACK, SO, OKAY, SO 842 00:37:19,040 --> 00:37:23,360 NEVER MIND, AS CAN YOU SEE, OVER 843 00:37:23,360 --> 00:37:28,280 HERE, THE--SO THE GLYCANS, 844 00:37:28,280 --> 00:37:30,920 OGLYCANS ACTUALLY ACTUALLY IS 845 00:37:30,920 --> 00:37:32,520 PRETTY ABUNDANTOT SURFACE, SO WE 846 00:37:32,520 --> 00:37:35,200 BASICALLY FOCUS ON THE GLYCANS 847 00:37:35,200 --> 00:38:18,480 OR ON THIS PROJECT--APPLY THESE 848 00:38:18,480 --> 00:38:19,640 STRUCTURES, USE THEM AS 849 00:38:19,640 --> 00:38:21,480 STANDARDS OF PROBES OR USE THAT 850 00:38:21,480 --> 00:38:24,120 AS BIOMARKERS SO THIS IS A 851 00:38:24,120 --> 00:38:26,840 SUCCESSFUL STORY ABOUT THIS 852 00:38:26,840 --> 00:38:30,960 WHICH IS A GLYCO PROTEIN AND USE 853 00:38:30,960 --> 00:38:43,200 THAT AS VACCINE DEVELOPMENT AND 854 00:38:43,200 --> 00:38:44,120 THERAPEUTICS FOR EXAMPLE, THESE 855 00:38:44,120 --> 00:39:10,760 CAN BE USED FOR DISEASES. 856 00:39:10,760 --> 00:39:12,720 --BUT CAN CAN BE REALLY 857 00:39:12,720 --> 00:39:14,920 COMLICATED HERE, BUT WE LOOK AT 858 00:39:14,920 --> 00:39:20,240 THIS IN DETAIL, THERE ARE OTHER 859 00:39:20,240 --> 00:39:22,280 COURSE GLYCANS AND SOME OTHER 860 00:39:22,280 --> 00:39:26,440 MINOR STRUCTURES OVER HERE, SO 861 00:39:26,440 --> 00:39:29,320 THEY'RE REALLY LOW ABUNDANCE. 862 00:39:29,320 --> 00:39:31,520 I MEAN DOUG TOLD YOU HOW TO 863 00:39:31,520 --> 00:39:33,880 SEPARATE THESE BUT FOR THE LOW 864 00:39:33,880 --> 00:39:35,400 ABUNDANCE WE THINK THESE WILL BE 865 00:39:35,400 --> 00:39:40,600 THE BETTER CHOICE, SO OF COURSE 866 00:39:40,600 --> 00:40:01,400 WE LOOK AT THESE CORE 867 00:40:01,400 --> 00:40:03,080 STRUCTURES,----SO, OF COURSE, 868 00:40:03,080 --> 00:40:04,400 FOR THE OLD SELECTING THE SAME 869 00:40:04,400 --> 00:40:07,760 THING, YOU HAVE THE COURSE, YOU 870 00:40:07,760 --> 00:40:14,440 HAVE MULTIPLE EPIEPITOPES, SO 871 00:40:14,440 --> 00:40:16,720 HERE IS BASICALLY IS IN-BOUND 872 00:40:16,720 --> 00:40:20,960 FOR THE USE HERE, THIS IS A LOT 873 00:40:20,960 --> 00:40:34,560 OF GLYCANS ON THE GLYCANS HERE. 874 00:40:34,560 --> 00:40:35,240 FOR THIS. 875 00:40:35,240 --> 00:40:39,240 AND OUR PROTEIN HERE IS 876 00:40:39,240 --> 00:40:43,080 OBVIOUSLY WE TRY TO USE CHEMICAL 877 00:40:43,080 --> 00:40:46,760 WAYS TO SYNTHESIZE THE REALLY 878 00:40:46,760 --> 00:40:47,800 CONSERVED CALL STRUCTURES, FOR 879 00:40:47,800 --> 00:40:49,200 EXAMPLE, WE HAVE THIS BEFORE, 880 00:40:49,200 --> 00:40:51,280 AND RIGHT NOTICE WE BASICALLY 881 00:40:51,280 --> 00:40:53,920 APPLY THESE METHODS OF COULD YOU 882 00:40:53,920 --> 00:40:58,840 SAYS, TO ODPLI KAN--KANAS. 883 00:40:58,840 --> 00:41:05,480 ONE OR 2 OR 3 SUGARS WHICH ARE 884 00:41:05,480 --> 00:41:07,680 RELATIVELY EASY TO SYNTHESIZE IN 885 00:41:07,680 --> 00:41:09,080 LARGE SCALE BUT CAN YOU SEE THE 886 00:41:09,080 --> 00:41:12,760 PROTEINS, AND THEN WE HAVE ALL 887 00:41:12,760 --> 00:41:15,200 THESE MATERIALS OR ENZYMES TO 888 00:41:15,200 --> 00:41:15,840 [INDISCERNIBLE]. 889 00:41:15,840 --> 00:41:18,840 AND LATER ON, WE WILL TRY TO USE 890 00:41:18,840 --> 00:41:24,120 THE SENSES TO SIMPLIFY SENSES. 891 00:41:24,120 --> 00:41:26,480 I WOULD JUST USE 1 SLIDE TO 892 00:41:26,480 --> 00:41:30,360 SUMMARIZE WHAT WE HAVE DONE NOW 893 00:41:30,360 --> 00:41:33,120 BASICALLY WE STARTED WITH THIS, 894 00:41:33,120 --> 00:41:35,720 WE DESIGNED THIS REALLY 895 00:41:35,720 --> 00:41:36,720 UNIVERSAL BUILDING BLOCK OVER 896 00:41:36,720 --> 00:41:39,520 HERE AND THEN WITH 2, THE FIRST 897 00:41:39,520 --> 00:41:41,240 STEPS WE CAN BASICALLY CONVERT 898 00:41:41,240 --> 00:41:47,800 THIS 1 TO ALL THE COURSE, A 899 00:41:47,800 --> 00:41:53,080 COURSE AND YIELDS FROM 51%, TO 900 00:41:53,080 --> 00:41:56,920 84%. 901 00:41:56,920 --> 00:41:59,720 SO THESE ARE ALL FOR THE SYNTH 902 00:41:59,720 --> 00:42:01,840 ASSESSING SOLID BASE, AND THEN, 903 00:42:01,840 --> 00:42:05,240 WE CAN ALSO PROTECT THAT FOR THE 904 00:42:05,240 --> 00:42:07,360 TPHREU RACING KAN--KANA SYNTHASE 905 00:42:07,360 --> 00:42:08,840 FOR THE WILD 2 OR 3 STEPS, 906 00:42:08,840 --> 00:42:28,040 USUALLY THE YIELD IS PRETTY 907 00:42:28,040 --> 00:42:28,240 GOOD. 908 00:42:28,240 --> 00:42:37,320 I WILL SKIP THIS, IT'S THE SAME 909 00:42:37,320 --> 00:42:39,240 THING--I LITTLE BIT UP SO WE 910 00:42:39,240 --> 00:42:42,440 HAVE ALL THESE STANDARD 911 00:42:42,440 --> 00:42:43,360 OPERATIONAL PROCEDURES 912 00:42:43,360 --> 00:42:46,640 AVAILABLE, SO WE HAVE PUT ALL 913 00:42:46,640 --> 00:42:48,320 THIS GLYCO CULTURES AND THESE 914 00:42:48,320 --> 00:42:50,640 CRITIC STEPS SO PEOPLE WANT TO 915 00:42:50,640 --> 00:42:53,760 REPRODUCE, TO REPEAT IT, IS THEY 916 00:42:53,760 --> 00:42:55,520 CAN REALLY FOLLOW THESE 917 00:42:55,520 --> 00:42:56,800 PROCEDURES IN THIS WEBSITE AND 918 00:42:56,800 --> 00:43:07,040 ALSO MY OWN GROUP WEBSITE. 919 00:43:07,040 --> 00:43:17,600 SO WE HAVE ENZYMES AND THIS 920 00:43:17,600 --> 00:43:20,840 ENZYMES AND EXTEND THESE TO THE 921 00:43:20,840 --> 00:43:29,240 NGLYCANS AND WE PEPTIDES AND 922 00:43:29,240 --> 00:43:32,560 ALSO ALSO THIS IS PRETTY 923 00:43:32,560 --> 00:43:34,080 EFFICIENT, SO WE COLLECTED 924 00:43:34,080 --> 00:43:52,520 ENZYMES AND LATER ON WE TRY TO 925 00:43:52,520 --> 00:43:56,600 ABOUT 5 OR 6--SO THIS AS 2 926 00:43:56,600 --> 00:44:00,840 BRANCHES IT BRANCH CAN EXTEND ON 927 00:44:00,840 --> 00:44:03,280 THE GLUNAC AND THIS CAN EXTEND 928 00:44:03,280 --> 00:44:03,840 ONED [INDISCERNIBLE]. 929 00:44:03,840 --> 00:44:16,440 SO WE CAN TEST THESE USING VERY 930 00:44:16,440 --> 00:44:18,760 SIMLE IF GIVES SOME ROBUST TOOLS 931 00:44:18,760 --> 00:44:22,520 TO REALLY RAPIDLY EXTEND MAYBE 1 932 00:44:22,520 --> 00:44:26,160 OR 2 CORE STRUCTURES CAN 933 00:44:26,160 --> 00:44:28,240 SYNTHESIZED CHEMICAL TO MAYBE 934 00:44:28,240 --> 00:44:29,480 DOZENS OR EVEN 10S OF 935 00:44:29,480 --> 00:44:33,600 STRUCTURES, I WILL GIVE YOU 1 936 00:44:33,600 --> 00:44:35,800 EXAMPLE HERE, SO, THE SENSES ARE 937 00:44:35,800 --> 00:44:37,000 DPLI KAN--KANAS SO AGAIN 938 00:44:37,000 --> 00:44:39,720 STARTING WITH THE BLOG ISSUING 939 00:44:39,720 --> 00:44:41,600 WE SYNTHESIZED A FEW CLAUSE 940 00:44:41,600 --> 00:44:45,880 CHEMICALLY AND WE COMBINE THIS 941 00:44:45,880 --> 00:44:48,040 WITH ANOTHER 13 ENZYME ADIOS AND 942 00:44:48,040 --> 00:44:49,760 IN THESE AUDYES WE WILL 943 00:44:49,760 --> 00:44:52,960 HAVE--EACH 1 WILL HAVE A 944 00:44:52,960 --> 00:44:55,880 SPECIFIC ENZYME WHICH CAN 945 00:44:55,880 --> 00:44:56,960 TRANSFER AND RECOGNIZE SINGLE 946 00:44:56,960 --> 00:44:59,120 ACCEPTABLE AND CAN CATALYZE THE 947 00:44:59,120 --> 00:45:01,840 SYNTHESIS OF A SINGLE GLYCO 948 00:45:01,840 --> 00:45:02,120 SITTIC BOND. 949 00:45:02,120 --> 00:45:11,680 SO THESE WE ALSO HAVE SUGAR 950 00:45:11,680 --> 00:45:12,520 NUCLEOTIDES AND I THINK THEY 951 00:45:12,520 --> 00:45:18,160 WILL TALK ABOUT THESE, THE 952 00:45:18,160 --> 00:45:19,800 COMMERCIALIZE THESE THEY ARE 953 00:45:19,800 --> 00:45:22,120 PRODUCE THESE IN REALLY LOW 954 00:45:22,120 --> 00:45:22,760 PRICES. 955 00:45:22,760 --> 00:45:24,840 SO 1 EXAMPLE HERE TO EXTEND THE 956 00:45:24,840 --> 00:45:32,160 CORE TO, SO I MEAN FROM 1 WE CAN 957 00:45:32,160 --> 00:45:34,720 DESIGN AND THIS CAN BE FURTHER 958 00:45:34,720 --> 00:45:35,840 EXTENDING FOR SURE, UP HERE. 959 00:45:35,840 --> 00:45:41,160 SO YOU CAN REALLY GET A LOT OF 960 00:45:41,160 --> 00:45:41,440 STRUCTURES. 961 00:45:41,440 --> 00:45:46,280 BUT AGAIN, SO [INDISCERNIBLE] 962 00:45:46,280 --> 00:45:49,840 BASED ON THE SPECIFICITY OF THE 963 00:45:49,840 --> 00:45:52,720 ENZYMES AND FOR EXAMPLE,--FOR 964 00:45:52,720 --> 00:45:54,360 THIS PAPER, WE SYNTHESIZED ABOUT 965 00:45:54,360 --> 00:45:57,120 80 OF THESE GLYCANS AND OF 966 00:45:57,120 --> 00:45:58,640 COURSE, FOR [INDISCERNIBLE] 967 00:45:58,640 --> 00:46:00,240 GLYCANS WE CAN START IT FROM 968 00:46:00,240 --> 00:46:02,000 BLOCKS HERE AND TO SYNTHESIZE 969 00:46:02,000 --> 00:46:04,320 MAYBE 30 OR MORE STRUCTURES OVER 970 00:46:04,320 --> 00:46:04,640 HERE. 971 00:46:04,640 --> 00:46:08,240 SO WE USE THESE TO DO A 972 00:46:08,240 --> 00:46:10,280 MICROARRAY AND TEST ACTIVITY BUT 973 00:46:10,280 --> 00:46:12,920 I WILL NOW SHOW HERE. 974 00:46:12,920 --> 00:46:16,400 AND WITH THAT, EVEN THOUGH THE 975 00:46:16,400 --> 00:46:18,240 ENZYMES AREENTIOUS FICIENT, I 976 00:46:18,240 --> 00:46:20,480 STILL NEED SOME EXPERTISE TO 977 00:46:20,480 --> 00:46:24,680 DEAL WITH THIS AND THERE'S A LOT 978 00:46:24,680 --> 00:46:26,040 THOSE THAT CAN TAKE EFFORTS, I 979 00:46:26,040 --> 00:46:30,000 THINK OUR GOAL AND THE GOAL OF 980 00:46:30,000 --> 00:46:31,040 THIS COMMUNITY, OF THESE COMMON 981 00:46:31,040 --> 00:46:33,560 FUND PROGRAM IS TRY TO SIMPLIFY 982 00:46:33,560 --> 00:46:37,800 THESE AND MAYBE IDEALLY TO DO 983 00:46:37,800 --> 00:46:38,840 AUTOMATIC SENSES, I THINK YOU 984 00:46:38,840 --> 00:46:40,200 LEARN FROM [INDISCERNIBLE] VERY 985 00:46:40,200 --> 00:46:43,680 SOON ABOUT THE CHEMICAL 986 00:46:43,680 --> 00:46:46,160 SYNTHESIS WHICH USES AUTOMATIC 987 00:46:46,160 --> 00:46:46,520 [INDISCERNIBLE]. 988 00:46:46,520 --> 00:46:50,760 SO WHEAT TRYING TO COMBINE THIS 989 00:46:50,760 --> 00:46:52,560 ENZYMATIC STUFF INTO THIS 990 00:46:52,560 --> 00:46:53,040 AUTOMATIC SYNTHESIS. 991 00:46:53,040 --> 00:46:55,880 SO WHAT WE DO IS WE USE THE 992 00:46:55,880 --> 00:46:57,560 SYNTHESIZER, WE PUT ALL THE 993 00:46:57,560 --> 00:47:01,160 REGIONS IN THE TUBES AND 994 00:47:01,160 --> 00:47:02,800 ENZYMES, NUCLEOTIDES AND OTHER 995 00:47:02,800 --> 00:47:05,240 IONS AND THEN WE CONJUGATED THE 996 00:47:05,240 --> 00:47:08,960 STARTING GLYCAN ON TO A RISEN OR 997 00:47:08,960 --> 00:47:11,160 SOMETHING INTO THIS VILE AND CAN 998 00:47:11,160 --> 00:47:15,920 PROGRAM IT TO DO THE SIPGHT SIS. 999 00:47:15,920 --> 00:47:17,160 [INDISCERNIBLE] LEADED THIS 1000 00:47:17,160 --> 00:47:23,040 PROJECT, THIS 1. 1001 00:47:23,040 --> 00:47:24,240 SO THE PROBLEM IS SO THE PROBLEM 1002 00:47:24,240 --> 00:47:27,040 IS THESE ARE NOT SOLID FACE, THE 1003 00:47:27,040 --> 00:47:29,240 SOLIDS WE USED FOR THE 60 SIS, 1004 00:47:29,240 --> 00:47:31,360 IT HAS A LITTLE [INDISCERNIBLE] 1005 00:47:31,360 --> 00:47:34,680 SO IT CAN INCREASE LOW INCAP 1006 00:47:34,680 --> 00:47:36,840 SIDITYY BUT ENZYMES LOOKS LIKE 1007 00:47:36,840 --> 00:47:38,560 THEY'RE NOT SO GOOD AT GOING 1008 00:47:38,560 --> 00:47:40,200 INTO THE [INDISCERNIBLE] BECAUSE 1009 00:47:40,200 --> 00:47:43,080 THEY'RE RELATIVELY LARGE, SO THE 1010 00:47:43,080 --> 00:47:45,800 EFFICIENCIES, WHAT WE DO OVER 1011 00:47:45,800 --> 00:47:49,120 THERE IS WE USE THE MATERIAL 1012 00:47:49,120 --> 00:47:50,320 HERE, SO AT LOW TEMPERATURE 1013 00:47:50,320 --> 00:47:53,000 WHICH ENZYME WILL WORK, OR 1014 00:47:53,000 --> 00:47:54,680 30-DEGREE TO THE SYNTHESIS AND 1015 00:47:54,680 --> 00:48:01,840 THEN IT WILL PRESS IT OUT. 1016 00:48:01,840 --> 00:48:03,280 AND THIS--THIS SYNTHESIZER, SO 1017 00:48:03,280 --> 00:48:06,440 WE HAVE 3 STEPS SO WE WILL 1018 00:48:06,440 --> 00:48:07,040 LOAD--FIRST LOAD THE TPHREU 1019 00:48:07,040 --> 00:48:09,840 RACING KAN--KANAS ON TO A RISEN, 1020 00:48:09,840 --> 00:48:12,040 ON TO A POLYMER AND THEN WE ADD 1021 00:48:12,040 --> 00:48:13,720 ALL THIS HERE, WITH THE PROGRAM 1022 00:48:13,720 --> 00:48:16,560 AND THEN WE CAN INCREASE THE 1023 00:48:16,560 --> 00:48:18,280 TEMPERATURE TO ROOM TEMPERATURE 1024 00:48:18,280 --> 00:48:21,920 OR MAYBE JUST A DEWITH THE 1025 00:48:21,920 --> 00:48:22,280 [INDISCERNIBLE]. 1026 00:48:22,280 --> 00:48:23,600 AND THEN AFTER DIRECTION TELL 1027 00:48:23,600 --> 00:48:27,000 INCREASE TEMPERATURE TO 1028 00:48:27,000 --> 00:48:27,400 [INDISCERNIBLE] TO 1029 00:48:27,400 --> 00:48:29,160 [INDISCERNIBLE] ENZYMES TO 1030 00:48:29,160 --> 00:48:31,480 REMOVE ALL THE SUGAR NUCLEOTIDES 1031 00:48:31,480 --> 00:48:33,840 OR OTHER STUFF AND WE CAN DO 1032 00:48:33,840 --> 00:48:36,560 THIS CYCLE BY CYCLE AND 1033 00:48:36,560 --> 00:48:40,440 SYNTHESIZE ALL THESE LITTLE BIT 1034 00:48:40,440 --> 00:48:40,880 COMPLEX STRUCTURES. 1035 00:48:40,880 --> 00:48:42,200 AND ALSO WHAT WEMENT TO DO IS WE 1036 00:48:42,200 --> 00:48:45,840 ALSO WANT TO DO THE GLYCO 1037 00:48:45,840 --> 00:48:47,240 PEPTIDE SYNTHESIZE, THIS THING 1038 00:48:47,240 --> 00:48:51,400 IS AGAIN, ENZYMES ARE NOT 1039 00:48:51,400 --> 00:48:53,840 COMPATIBLE WITH SOLID PHASE, AND 1040 00:48:53,840 --> 00:48:56,200 WHEN AGAIN BECAUSE THEY 1041 00:48:56,200 --> 00:48:57,840 CANNOT--I MEAN SNEAK INTO THE 1042 00:48:57,840 --> 00:48:59,520 SMALL PORES, RIGHT NOW WE CHAIN 1043 00:48:59,520 --> 00:49:01,640 THAT INTO AN ACIDIC BOWL. 1044 00:49:01,640 --> 00:49:03,600 SO THE CAP SID HERE, SO 1045 00:49:03,600 --> 00:49:05,200 BASICALLY WE ONLY HAVE THE DPLI 1046 00:49:05,200 --> 00:49:05,880 KAN--KANAS LOADING ON THE 1047 00:49:05,880 --> 00:49:07,480 SURFACE OF THE BALL. 1048 00:49:07,480 --> 00:49:09,680 THE LOADING CAP SID IS CLEARLY 1049 00:49:09,680 --> 00:49:11,560 PRETTY LOW BUT IT'S BETTER FOR 1050 00:49:11,560 --> 00:49:12,440 ENZYMES TO WORK, SO LAWN MOWER 1051 00:49:12,440 --> 00:49:15,200 WE DO IS BASICALLY WE--WE HAVE 2 1052 00:49:15,200 --> 00:49:17,240 SIDES, SO 1 SIDE ON THE LEFT, WE 1053 00:49:17,240 --> 00:49:20,320 WILL DO THE SOLID FACE PEPTIDE 1054 00:49:20,320 --> 00:49:22,280 SYNTHIZE, EVERYTHING IS SAME TO 1055 00:49:22,280 --> 00:49:24,200 THE TRADITIONAL SYNTHESIS, THE 1056 00:49:24,200 --> 00:49:25,640 ONLY DIFFERENCE IS WE WILL 1057 00:49:25,640 --> 00:49:27,240 CHANGE THIS TO [INDISCERNIBLE], 1058 00:49:27,240 --> 00:49:31,960 AND AFTER WE HAVE ALL THESE 1059 00:49:31,960 --> 00:49:33,120 OORK, SO WE WILL INTRODUCE THIS 1060 00:49:33,120 --> 00:49:35,920 YEAR AND WE WILL GET SOME 1061 00:49:35,920 --> 00:49:36,840 PEPTIDES WITH THE 1062 00:49:36,840 --> 00:49:37,240 [INDISCERNIBLE]. 1063 00:49:37,240 --> 00:49:41,680 AND WE WILL TRANSFER THAT INTO 1064 00:49:41,680 --> 00:49:44,320 AN AQUEOUS SOLUTION, AND ALL OF 1065 00:49:44,320 --> 00:49:46,480 THESE CAN BE [INDISCERNIBLE] AND 1066 00:49:46,480 --> 00:49:50,880 NORMALLY WE HAVE THESE, WE HAVE 1067 00:49:50,880 --> 00:49:54,000 ALL THESE OTHER STRUCTURES WITH 1068 00:49:54,000 --> 00:49:56,680 PEPTIDES WE JUST DO THE PROGRAM 1069 00:49:56,680 --> 00:50:04,600 AND THE MACHINE WILL DO ALL THE 1070 00:50:04,600 --> 00:50:05,040 STUFF. 1071 00:50:05,040 --> 00:50:07,240 SO TO SUMMARIZE, WE HAVE ALL 1072 00:50:07,240 --> 00:50:09,040 THESE AVAILABLE ONLINE, HOW TO 1073 00:50:09,040 --> 00:50:10,400 CALCULATE THROUGH THE COURSE AND 1074 00:50:10,400 --> 00:50:13,200 ENZYMES SO WE TRY TO CLICK INTO 1075 00:50:13,200 --> 00:50:14,960 THESE LINKS HERE, WE SEE HOW TO 1076 00:50:14,960 --> 00:50:16,360 EXPRESS THESE ENZYMES HOW TO 1077 00:50:16,360 --> 00:50:19,360 PURIFY, HOW TO CHOOSE THE ASSAY, 1078 00:50:19,360 --> 00:50:21,280 AND WHAT IS SPECIFICITY OF THE 1079 00:50:21,280 --> 00:50:24,040 ENZYMES, SO THIS IS REALLY A 1080 00:50:24,040 --> 00:50:24,680 DETAILED INFORMATION HERE. 1081 00:50:24,680 --> 00:50:30,960 AND OF COURSE, THE VALIDATION, 1082 00:50:30,960 --> 00:50:32,640 DO THIS VALIDATION AND WE ALSO 1083 00:50:32,640 --> 00:50:35,760 HAVE COLLABORATIONS, SO THEY GOT 1084 00:50:35,760 --> 00:50:38,080 OUR CLOTHES AND OUR PEPTIDES 1085 00:50:38,080 --> 00:50:43,800 THERE THAT WORK. 1086 00:50:43,800 --> 00:50:45,280 [INDISCERNIBLE] WE PUBLISHED 1087 00:50:45,280 --> 00:50:46,680 THESE PAPERS. 1088 00:50:46,680 --> 00:50:48,720 AND FINALLY, I WILL SKIP ALL 1089 00:50:48,720 --> 00:50:50,440 THESE PAPERS, SO FINALLY, I 1090 00:50:50,440 --> 00:50:53,280 WOULD LIKE TO THANK DOUG'S GROUP 1091 00:50:53,280 --> 00:50:55,280 FOR DOING THE CROSS VALIDATION 1092 00:50:55,280 --> 00:50:57,960 AND THE SHARE THE ENZYME AND THE 1093 00:50:57,960 --> 00:50:59,440 CYTOTRANSFER DISEASE AND THIS IS 1094 00:50:59,440 --> 00:51:03,840 A GROUP OVER HERE AND ALSO MOST 1095 00:51:03,840 --> 00:51:05,160 IMPORTANTLY THE GLYCO COMMON 1096 00:51:05,160 --> 00:51:07,400 FUND TO SPOIL US AND RIGHT NOW 1097 00:51:07,400 --> 00:51:10,080 WE HAVE THIS GROUND TO 1098 00:51:10,080 --> 00:51:12,760 SYNTHESIZE PEPTIDES AND WE HAVE 1099 00:51:12,760 --> 00:51:17,480 THIS SBIR TOGETHER WITH 1100 00:51:17,480 --> 00:51:18,640 [INDISCERNIBLE] TO SYNTHESIZE 1101 00:51:18,640 --> 00:51:27,040 MUSEIN PEPTIDES, SO THAT'S ALL. 1102 00:51:27,040 --> 00:51:28,000 THANK YOU. 1103 00:51:28,000 --> 00:51:29,240 [ APPLAUSE ] 1104 00:51:29,240 --> 00:51:29,920 >> THANK YOU. 1105 00:51:29,920 --> 00:51:37,200 SO OUR NEXT SPEAKER TODAY IS BY 1106 00:51:37,200 --> 00:51:46,800 DR. POLIZZI, FROM CHEMILY. 1107 00:51:46,800 --> 00:51:49,960 >> IT'S A PLEASURE TO SPEAK WITH 1108 00:51:49,960 --> 00:51:50,640 YOU TODAY. 1109 00:51:50,640 --> 00:51:51,800 SO THE PREVIOUS SPEAKER VS BEEN 1110 00:51:51,800 --> 00:51:55,240 SO KIND TO INTRODUCE A LOT OF 1111 00:51:55,240 --> 00:51:56,400 IMPORTANT SUGARS TO YOU SO 1112 00:51:56,400 --> 00:51:59,960 THEY'VE DONE GROUND FOR FOR ME, 1113 00:51:59,960 --> 00:52:02,200 AND HOW SHOWS SUGARS ARE AND 1114 00:52:02,200 --> 00:52:03,640 WHERE THEY'RE COMING FROM, FOR 1115 00:52:03,640 --> 00:52:11,640 TODAY, SO I SPEAKING TO YOU, WHY 1116 00:52:11,640 --> 00:52:13,040 CHEMILY, LLC, AND WE ARE 1117 00:52:13,040 --> 00:52:15,240 SPEAKING TO YOU ABOUT THESE 1118 00:52:15,240 --> 00:52:16,400 REAGENTS AS THE PREVIOUS 1119 00:52:16,400 --> 00:52:18,640 SPEAKERS AS WELL, SO I WANT TO 1120 00:52:18,640 --> 00:52:19,840 START WITH WITH ACKNOWLEDGMENTS 1121 00:52:19,840 --> 00:52:21,640 AND HE IS THE GROUP THAT WAS 1122 00:52:21,640 --> 00:52:23,720 UNDER GEORGE WONG AT THE GEORGIA 1123 00:52:23,720 --> 00:52:24,640 STATE UNIVERSITY HAVE REALLY 1124 00:52:24,640 --> 00:52:28,720 DEVELOPED A LOT OF THE METHODS 1125 00:52:28,720 --> 00:52:30,320 AND DONE A LOT OF CLONING AND 1126 00:52:30,320 --> 00:52:31,920 ENZYMES ARE GOING TO BE 1127 00:52:31,920 --> 00:52:33,320 IMPORTANT FOR WHAT I'M SPEAKING 1128 00:52:33,320 --> 00:52:35,200 TO YOU ABOUT RIGHT NOW AND THEN 1129 00:52:35,200 --> 00:52:36,600 WE'VE TAKEN SOME OF THEIR GREAT 1130 00:52:36,600 --> 00:52:38,520 TALENT FROM THAT LAB, AND 1131 00:52:38,520 --> 00:52:50,280 ACTUALLY MOVE THEM OVER INTO 1132 00:52:50,280 --> 00:52:51,320 CHEMILY, AND ALL COLLEAGUES HAD 1133 00:52:51,320 --> 00:52:53,120 BEEN AT THAT LAB IN THAT STATE. 1134 00:52:53,120 --> 00:52:55,600 I ALSO WANT TO THANK THE FUNDING 1135 00:52:55,600 --> 00:52:56,880 AS WELL, AND REITERATE FROM 1136 00:52:56,880 --> 00:52:58,240 YESTERDAY IT WAS INTERESTING AND 1137 00:52:58,240 --> 00:52:59,880 NOVEL FOR THE COMONT COMMON FUND 1138 00:52:59,880 --> 00:53:01,880 TO PROVIDE THEIR OWN SBIR AND WE 1139 00:53:01,880 --> 00:53:03,840 WERE FUNDED UNDER THE NEW 1140 00:53:03,840 --> 00:53:06,360 TECHNOLOGIES FOR THE FLIEK O 1141 00:53:06,360 --> 00:53:07,200 SCIENCES SOLICITATION. 1142 00:53:07,200 --> 00:53:08,840 SO JUST A LITTLE THOUGHT 1143 00:53:08,840 --> 00:53:12,600 EXPERIMENT, SO MAYBE A BIT ABOUT 1144 00:53:12,600 --> 00:53:15,320 YOUR EXPERIENCES SO WOULD YOU 1145 00:53:15,320 --> 00:53:17,600 LIKE YOUR UNDERGRADS MAKE A GTP 1146 00:53:17,600 --> 00:53:17,840 SOLUTION? 1147 00:53:17,840 --> 00:53:18,680 MAYBE SO. 1148 00:53:18,680 --> 00:53:20,640 KNOWING THEY WILL SPILL SOME, 1149 00:53:20,640 --> 00:53:21,840 WE'VE HEARD HEARD FROM NIKI THAT 1150 00:53:21,840 --> 00:53:24,400 WE ARE TRYING TO GET THESE INTO 1151 00:53:24,400 --> 00:53:26,680 OUR UNDERGRADUATES AND OUR NOVEL 1152 00:53:26,680 --> 00:53:27,240 SCIENTISTS HANDS, KNOWING 1153 00:53:27,240 --> 00:53:28,920 THEY'RE GOING TO RUN PC R 1154 00:53:28,920 --> 00:53:30,400 EXPERIMENTS, IT'S NOT ABOUT 1155 00:53:30,400 --> 00:53:31,400 MAKING SLIEWGDZS, IT'S ABOUT 1156 00:53:31,400 --> 00:53:34,240 WHAT YOU WILL DO WITH WITH THEM 1157 00:53:34,240 --> 00:53:35,640 AND OVER AND ORDER AGAIN AND 1158 00:53:35,640 --> 00:53:37,440 DEVELOP THE SKILLS AND WE SWITCH 1159 00:53:37,440 --> 00:53:39,360 THAT OVER TO FLIEK O EXPNS AND 1160 00:53:39,360 --> 00:53:41,520 THIS MAKES YOU MORE NERVOUS, 1161 00:53:41,520 --> 00:53:44,080 WILL YOU LET THEM MAKE GDP 1162 00:53:44,080 --> 00:53:45,400 SOLUTIONS OR WILL YOU DO THAT 1163 00:53:45,400 --> 00:53:46,360 KNOWING THEY WILL SPILL SOME OF 1164 00:53:46,360 --> 00:53:48,680 IT AND YOU MIGHT BE LOSING A 1165 00:53:48,680 --> 00:53:50,640 POST DOC'S WORTH OF WORK OR YOU 1166 00:53:50,640 --> 00:53:53,600 GOING TO LET THEM MOVE TO TO THE 1167 00:53:53,600 --> 00:53:54,240 GLYCOSYLATION EXPERIMENT WHICH 1168 00:53:54,240 --> 00:53:56,440 IS THE IMPORTANT PART AND RERUN 1169 00:53:56,440 --> 00:53:57,840 THAT MULTIPLE TIMES,IME NOT SURE 1170 00:53:57,840 --> 00:54:06,040 THAT'S PART OF 1 OF YOUR LABS SO 1171 00:54:06,040 --> 00:54:07,240 IF YOU'RE HEART GOT PUMPING AND 1172 00:54:07,240 --> 00:54:13,640 YOU WILL SAY YES, THEN THERE'S 1173 00:54:13,640 --> 00:54:15,320 SPECIAL OPPORTUNITY HERE TO COME 1174 00:54:15,320 --> 00:54:18,240 IN AND DESIGN THE SUGARS AND LOW 1175 00:54:18,240 --> 00:54:19,240 TO BE AFARREDDED AND SOMETHING 1176 00:54:19,240 --> 00:54:20,120 THAT WILL KEEP FRUIT FLIES 1177 00:54:20,120 --> 00:54:22,560 HAVING A HEART ATTACK IF THAT 1178 00:54:22,560 --> 00:54:24,640 HAPPENS, SO OUR GRANT THEN WAS 1179 00:54:24,640 --> 00:54:27,000 TO MAKE THESE NUCLEOTIDE SUGAR 1180 00:54:27,000 --> 00:54:29,080 BLOCKS, SINCE THESE WILL BE USED 1181 00:54:29,080 --> 00:54:31,520 BY ENZYMES TO BUILD THESE, BUT 1182 00:54:31,520 --> 00:54:32,480 OTHER PRESENTER VS PUT UP THERE 1183 00:54:32,480 --> 00:54:34,000 ON THE SCREEN, AS A SPECIAL 1184 00:54:34,000 --> 00:54:35,320 ORGANIZATIONS NIEGZ EGG, WE ARE 1185 00:54:35,320 --> 00:54:36,400 INTERESTED IN SERVING 1186 00:54:36,400 --> 00:54:36,720 OPPORTUNITIES. 1187 00:54:36,720 --> 00:54:38,480 WE WANT TO MAKE THE PIs 1188 00:54:38,480 --> 00:54:40,080 COMFORTABLE THAT THEY WILL BE 1189 00:54:40,080 --> 00:54:42,240 ABLE TO GET A STABLE SUPPLY OF 1190 00:54:42,240 --> 00:54:45,000 THESE REAGENTS AND THEN ALSO BE 1191 00:54:45,000 --> 00:54:46,200 ABLE ABLE TO ENABLE THEM TO DID 1192 00:54:46,200 --> 00:54:47,440 THE EXPERIMENTS THEY WANT TO DO 1193 00:54:47,440 --> 00:54:49,680 WHERE THEY'RE NOT SPEND 1194 00:54:49,680 --> 00:54:51,120 THAGOREAN 70-75% OF THEST JUST 1195 00:54:51,120 --> 00:54:52,200 MAKING THOSE STARTING MATERIALS, 1196 00:54:52,200 --> 00:54:54,720 AND THEN ALSO AS A COMMERCIAL 1197 00:54:54,720 --> 00:54:56,040 ORGANIZATION, WE HAVE 1198 00:54:56,040 --> 00:54:56,720 ASPIRATIONAL GOALS, WE'RE 1199 00:54:56,720 --> 00:54:58,040 INTERESTED IN TRYING TO 1200 00:54:58,040 --> 00:55:00,560 FACILITATE KIND OF THE NEXT 1201 00:55:00,560 --> 00:55:02,200 GENERATION EXPERIMENTS WHERE 1202 00:55:02,200 --> 00:55:03,640 YOU'RE NOT WORRIED ABOUT THE 1203 00:55:03,640 --> 00:55:04,640 STARTING MATERIALS, YOU ARE 1204 00:55:04,640 --> 00:55:06,640 REALLY THINKING ABOUT THE ROOL 1205 00:55:06,640 --> 00:55:09,040 SCIENCE INSTEAD OF THE NUTS AND 1206 00:55:09,040 --> 00:55:10,840 BOLTS OF WHERE YOUR SUPPLIES ARE 1207 00:55:10,840 --> 00:55:11,160 COMING FROM. 1208 00:55:11,160 --> 00:55:13,080 SO THIS WAS A PARTNERSHIP 1209 00:55:13,080 --> 00:55:16,360 BUILDING OUT OF THE UO1 THAT WE 1210 00:55:16,360 --> 00:55:21,240 HEARD FROM FROM DR. LI, AND THEY 1211 00:55:21,240 --> 00:55:23,360 STARTED IN 2015 WITH THE CORE 1212 00:55:23,360 --> 00:55:25,520 SYNTHESIS, AND THE CORE OF HIS 1213 00:55:25,520 --> 00:55:28,040 TALK WAS ABOUT THAT CSY 1214 00:55:28,040 --> 00:55:28,320 EESYSTEM. 1215 00:55:28,320 --> 00:55:30,440 SO WHAT WE DID WAS A COUPLE 1216 00:55:30,440 --> 00:55:33,040 YEARS INTO THAT UO1 WAS WE 1217 00:55:33,040 --> 00:55:34,240 STARTED COMMUNICATING WITH THEIR 1218 00:55:34,240 --> 00:55:35,440 GROUP, THEY'RE GIVING US 1219 00:55:35,440 --> 00:55:36,440 INFORMATION ENZYMES WE'RE 1220 00:55:36,440 --> 00:55:37,440 PUTTING TOGETHER A COMMERCIAL 1221 00:55:37,440 --> 00:55:39,880 COLLECTION OF THOSE ENZYMES, 1222 00:55:39,880 --> 00:55:41,160 PROTOCOLS, BEING OPTIMIZED, 1223 00:55:41,160 --> 00:55:43,240 DEVELOPING PRODUCTS IN AN EVENT 1224 00:55:43,240 --> 00:55:44,040 DIVERSIFYING SOME OF THOSE 1225 00:55:44,040 --> 00:55:46,240 PROWCTDS AND AT THAT POINT WE 1226 00:55:46,240 --> 00:55:49,640 CAN FEEDBACK THEM BACK INTO THAT 1227 00:55:49,640 --> 00:55:51,160 ORIGINAL UO1 AND WE CAN HELP 1228 00:55:51,160 --> 00:55:52,560 THEM OUT, THEY CAN HELP US OUT. 1229 00:55:52,560 --> 00:55:54,680 PART OF THE REGION THEY 1230 00:55:54,680 --> 00:55:56,320 SEQUENCED THESE TALKS THIS WAY 1231 00:55:56,320 --> 00:55:58,040 IS IF YOU'RE OUT THERE, IF 1232 00:55:58,040 --> 00:56:00,240 YOU'RE THINK BEING THE TYPES OF 1233 00:56:00,240 --> 00:56:01,880 PROWCTDS YOU FLEED AND THOSE 1234 00:56:01,880 --> 00:56:07,440 GRANTS CAN YOU PARTNER UP WITH 1235 00:56:07,440 --> 00:56:08,200 THAT COMMERCIAL ORGANIZATION, 1236 00:56:08,200 --> 00:56:10,640 NOT ONLY FOR YOUR LAB BUT ALSO 1237 00:56:10,640 --> 00:56:11,520 FOR OTHER SCIENTISTS WANTING TO 1238 00:56:11,520 --> 00:56:14,240 WORK IN THE FIELD. 1239 00:56:14,240 --> 00:56:17,080 SO WE COMMERCIALIZE 11 COMMON 1240 00:56:17,080 --> 00:56:19,040 NUCLEOTIDE SUGARS. 1241 00:56:19,040 --> 00:56:23,080 THEY'RE COMMON TO MAMMALIAN 1242 00:56:23,080 --> 00:56:24,640 SYSTEMS, AND THESE WERE NOT 1243 00:56:24,640 --> 00:56:26,960 AVAILABLE IN 2017, SO WE'RE 1244 00:56:26,960 --> 00:56:32,560 ENDING UP WITH 7 UDP SUGARS, 2 1245 00:56:32,560 --> 00:56:35,000 GDP SUGARS AND 2 CMP, SO WHAT'S 1246 00:56:35,000 --> 00:56:35,720 COMMON ABOUT THESE REACTION 1247 00:56:35,720 --> 00:56:37,440 SYSTEM WE LIKE TO START WITH 1248 00:56:37,440 --> 00:56:39,200 ENZYMES FROM THE SALVAGE 1249 00:56:39,200 --> 00:56:41,920 PATHWAYS, WE'RE TRYING TO TAKE 1250 00:56:41,920 --> 00:56:45,240 SOME STARTING VERY CHEAP 1251 00:56:45,240 --> 00:56:45,800 SUBSTRATES, AND THEN FIRST 1252 00:56:45,800 --> 00:56:47,360 FOSTER NURSED FARALATE THEM WITH 1253 00:56:47,360 --> 00:56:49,360 A SUGAR KINASE AND THEN WE GO 1254 00:56:49,360 --> 00:56:51,040 AHEAD AND ACTIVATE THEM WITH A 1255 00:56:51,040 --> 00:56:51,720 SECOND ENZYME. 1256 00:56:51,720 --> 00:56:53,880 THESE REACTIONS ARE ALSO 1257 00:56:53,880 --> 00:56:55,040 INVOLVING AN ALCO LINE FOSTER 1258 00:56:55,040 --> 00:56:56,160 NURSED FOCUSED ON TAIS INHIBITOR 1259 00:56:56,160 --> 00:56:57,640 WHICH WILL PUSH THIS FORWARD, 1260 00:56:57,640 --> 00:56:59,400 TRY TO OPTIMIZE THE AMOUNT OF 1261 00:56:59,400 --> 00:57:01,160 PRODUCT WE CAN GET AND SPEED UP 1262 00:57:01,160 --> 00:57:03,440 REACTIONS, IF YOU HAVE SOMETHING 1263 00:57:03,440 --> 00:57:05,080 LIKE GDP GLUCOSE THAT YOU'RE 1264 00:57:05,080 --> 00:57:06,840 INTERESTED IN, IT HAS CHEAP 1265 00:57:06,840 --> 00:57:09,120 STARTING MATERIALS, SO CAN YOU 1266 00:57:09,120 --> 00:57:12,160 START HERE AS OTO THESE 1 POT 1267 00:57:12,160 --> 00:57:12,840 MULTIENZYME SYSTEMS. 1268 00:57:12,840 --> 00:57:16,840 SO I WANT TO GIVE YOU A CASE 1269 00:57:16,840 --> 00:57:18,640 STUDY, SO UDP, MY FAVORITE 1270 00:57:18,640 --> 00:57:22,560 NUCLEOTIDE SUGAR IS THE 1271 00:57:22,560 --> 00:57:23,680 ACTIVATED XYLOSE, WHERE IT'S 1272 00:57:23,680 --> 00:57:25,960 ATTACHED TO SOMETHING LIKE NOTCH 1273 00:57:25,960 --> 00:57:26,760 SIGNALING RECEPTORS, LARGE, 1274 00:57:26,760 --> 00:57:27,760 THINGS LIKE THAT, THIS IS ALSO 1275 00:57:27,760 --> 00:57:30,480 GOING TO BE THE INITIATION STEP 1276 00:57:30,480 --> 00:57:33,240 FOR CORE PROTEIN LINKAGE TO 1277 00:57:33,240 --> 00:57:34,240 MULTIPLE CLASSES OF PROTEIN 1278 00:57:34,240 --> 00:57:35,840 COMPLEXIO GLYCANS, THAT TEST HAS 1279 00:57:35,840 --> 00:57:39,080 TO GO ON BEFORE THE REST OF THE 1280 00:57:39,080 --> 00:57:40,040 TETRASACCHARIDE LINKER CAN GO 1281 00:57:40,040 --> 00:57:42,520 ON, IN CELLS IT WILL BE PRODUCED 1282 00:57:42,520 --> 00:57:45,840 IN THIS DE NOVO PATHWAY, THE 1283 00:57:45,840 --> 00:57:48,200 UDP, THEN ON THE SILOS, THIS NOT 1284 00:57:48,200 --> 00:57:50,040 THE WAY YOU WANT TO MAKE THIS IN 1285 00:57:50,040 --> 00:57:50,640 YOUR LAB. 1286 00:57:50,640 --> 00:57:52,560 THERE'S A REASON WHY THESE ARE 1287 00:57:52,560 --> 00:57:54,520 DID IN DIFFERENT COMPARTMENTS 1288 00:57:54,520 --> 00:57:59,640 INSIDE THE CELL, NADH WILL 1289 00:57:59,640 --> 00:58:01,200 INHIBIT UDH, IT WILL BE 1290 00:58:01,200 --> 00:58:02,040 REGULATED FOR THIS, SO THERE 1291 00:58:02,040 --> 00:58:04,480 NEEDING TO BE A BETTER WAY TO 1292 00:58:04,480 --> 00:58:05,640 MAKE THREEZ SUGARS. 1293 00:58:05,640 --> 00:58:06,960 FROM THAT PREVIOUS VERY BUSY 1294 00:58:06,960 --> 00:58:09,120 SLIDE, I TOLD YOU WE WANT TO 1295 00:58:09,120 --> 00:58:19,760 LOOK AT SALVAGE PATHWAYS SO 1296 00:58:19,760 --> 00:58:21,440 STARTING FROM FOZ FORALATION 1 1297 00:58:21,440 --> 00:58:23,200 POSITION, AND THEN EDITING IT IN 1298 00:58:23,200 --> 00:58:23,400 TIME. 1299 00:58:23,400 --> 00:58:26,680 SO IF WE WANT TO GET TO XYLOSRKS 1300 00:58:26,680 --> 00:58:28,840 E, WE WANT TO UTILIZE THIS 1301 00:58:28,840 --> 00:58:29,080 PATHWAY. 1302 00:58:29,080 --> 00:58:31,360 WE ARE ALSO INTERESTED IN THE 1303 00:58:31,360 --> 00:58:32,840 SYNTHESIS, WHERE WE CAN BI PASS 1304 00:58:32,840 --> 00:58:34,240 INTENDED THE FIRST STEP, AND THE 1305 00:58:34,240 --> 00:58:36,840 1, AND THEN SEE IF WE CAN 1306 00:58:36,840 --> 00:58:40,680 ENZYMATICALLY PUSH IT INTO THOSE 1307 00:58:40,680 --> 00:58:41,120 FINAL PRODUCTS. 1308 00:58:41,120 --> 00:58:43,120 SO WITH THAT, WE STARTED OFF 1309 00:58:43,120 --> 00:58:45,440 WITH 2 STEPS, TO GET US TO THE 1310 00:58:45,440 --> 00:58:47,640 SUGAR PHOSPHATE, WE HAVE A 1311 00:58:47,640 --> 00:58:50,600 [INDISCERNIBLE] HERE AND WE HAVE 1312 00:58:50,600 --> 00:58:54,520 THIS WITH THE RABBINOSE AND 1313 00:58:54,520 --> 00:58:56,400 XYLOSE, AND WE LOOK AT THE 1314 00:58:56,400 --> 00:58:59,720 CHEMICAL SYNTHESIS, WE END UP 1315 00:58:59,720 --> 00:59:03,040 WITH A ANAMERRIC MIXTURE, AND WE 1316 00:59:03,040 --> 00:59:04,920 HAVE EVIDENCE THAT HAPPENS AFTER 1317 00:59:04,920 --> 00:59:06,240 THE CHEMICAL PROCESSING BUT THEN 1318 00:59:06,240 --> 00:59:07,440 AFTER WE INTRODUCE THE ENZYME, 1319 00:59:07,440 --> 00:59:11,320 WE WILL SELECT FOR THE 1320 00:59:11,320 --> 00:59:12,960 BIOLOGICALLY RELEVANT ANAMERAND 1321 00:59:12,960 --> 00:59:16,960 WE ARE PRODUCING ENTIRELY 1322 00:59:16,960 --> 00:59:18,280 ALPHA-D XYLOSE IN THIS CASE. 1323 00:59:18,280 --> 00:59:25,640 BEING WE'RE A COMMERCIAL 1324 00:59:25,640 --> 00:59:27,200 ORGANIZATION, WE'RE INTERESTED 1325 00:59:27,200 --> 00:59:28,760 IN IN THIS MORE FEASIBLE AT 1326 00:59:28,760 --> 00:59:31,440 SCALE SO WE'VE BEEN ABLE TO 1327 00:59:31,440 --> 00:59:32,760 SCREEN ADDITIONAL ENZYMES AND IN 1328 00:59:32,760 --> 00:59:35,400 SOME CASES WOO CAN END UP WITH 1329 00:59:35,400 --> 00:59:37,480 MULTIPLE HITS, IN SOME CASES 1330 00:59:37,480 --> 00:59:39,280 JUST A SINGLE HIT BUT SOME OF US 1331 00:59:39,280 --> 00:59:41,440 TO OPTIMIZE THE INITIAL 1332 00:59:41,440 --> 00:59:42,840 ANALYTICAL PROOF OF CONCEPT, 1333 00:59:42,840 --> 00:59:47,120 THIS COMING OUT OF THE ACADEMIC 1334 00:59:47,120 --> 00:59:47,320 LABS. 1335 00:59:47,320 --> 00:59:49,200 BEYOND THAT WE'VE BEEN ABLE TO 1336 00:59:49,200 --> 00:59:51,520 TAKE THAT SAME TYPE OF STRATEGY 1337 00:59:51,520 --> 00:59:53,240 WHERE WE START OFF WITH THE 1338 00:59:53,240 --> 00:59:54,880 STARTING MATERIALS AND GO IN AND 1339 00:59:54,880 --> 00:59:56,640 THEN MODIFY THEM WITH ENZYMES SO 1340 00:59:56,640 --> 00:59:58,440 WE CAN CLEAN UP ANY OF THE 1341 00:59:58,440 --> 00:59:59,560 ANAMERRIC HAD OR OTHER MIXTURES 1342 00:59:59,560 --> 01:00:01,840 THAT WOULD HAVE COME FROM THOSE 1343 01:00:01,840 --> 01:00:03,160 SYNTHETIC PROCESSES, SO ALMOST A 1344 01:00:03,160 --> 01:00:06,360 CLEANING STEP AS WELL, SO ADD IN 1345 01:00:06,360 --> 01:00:08,400 KD--SALLY WHARKSAD IN CTP, THESE 1346 01:00:08,400 --> 01:00:11,160 ENZYMES AND THEN WE'RE ABLE TO 1347 01:00:11,160 --> 01:00:14,640 MAKE THE CMP ACID DERIVATIVES. 1348 01:00:14,640 --> 01:00:16,680 NOW THE BAM HALF OF THIS LIST IS 1349 01:00:16,680 --> 01:00:18,880 THINGS WE PRODUCED IN THE 1350 01:00:18,880 --> 01:00:20,160 HUNDRED MILLIGRAM SCALES, AND IN 1351 01:00:20,160 --> 01:00:22,920 YEAR 4 OF THE FAST TRACK AND IN 1352 01:00:22,920 --> 01:00:25,200 THE TOP HALF WE PREPARED THE 1353 01:00:25,200 --> 01:00:27,280 SUGARS AND IN THIS FINAL FIFTH 1354 01:00:27,280 --> 01:00:29,480 YEAR, WE'RE WORKING UP THE 1355 01:00:29,480 --> 01:00:30,840 SCALES OF THIS PRODUCTION ON 1356 01:00:30,840 --> 01:00:33,080 THIS AS L. SO AS WELL AS TRYING 1357 01:00:33,080 --> 01:00:35,040 TO BE COMMERCIALLY VIABLE, WE 1358 01:00:35,040 --> 01:00:36,480 ARE THINKING ABOUT THOSE 1359 01:00:36,480 --> 01:00:37,960 UNDERGRADUATES CAN WE PUT THINGS 1360 01:00:37,960 --> 01:00:39,880 IN KIDS, CAN WE GIVE THEM THE 1361 01:00:39,880 --> 01:00:40,960 OPPORTUNITY TO MAKE NUCLEOTIDE 1362 01:00:40,960 --> 01:00:42,440 SUGAR RIGHT THERE IN WHATEVER 1363 01:00:42,440 --> 01:00:43,480 REACTION THEY'RE DOING SO 1364 01:00:43,480 --> 01:00:45,040 EVERYTHING THEY NEED TO GET THAT 1365 01:00:45,040 --> 01:00:47,240 DONE, THEY CAN ADD THEIR OWN 1366 01:00:47,240 --> 01:00:48,680 TRANSFER ACE AND SEE IF IT'S 1367 01:00:48,680 --> 01:00:50,760 APPLIED TO THEIR PRODUCT OF 1368 01:00:50,760 --> 01:00:51,680 INTEREST, DERIFFITIZATION, SO IF 1369 01:00:51,680 --> 01:00:53,440 HAVE YOU YOUR OWN SUGAR, CAN YOU 1370 01:00:53,440 --> 01:00:55,600 POP THAT SUGAR IN THERE 1371 01:00:55,600 --> 01:00:58,120 ALONGSIDE WHAT WOULD THEN BE A 1372 01:00:58,120 --> 01:00:59,440 POSITIVE CONTROL WITH THE 1373 01:00:59,440 --> 01:01:00,800 STARTING SUGAR WE SUPPLIED IN 1374 01:01:00,800 --> 01:01:01,600 THE KIDS. 1375 01:01:01,600 --> 01:01:02,840 SAMPLER BACKS IF YOU GO TO 1376 01:01:02,840 --> 01:01:06,640 SIGMA, CAN YOU BUY A WHOLE 1377 01:01:06,640 --> 01:01:07,840 SEQUENCE OF NUCLEOTIDE TRI 1378 01:01:07,840 --> 01:01:11,920 POSFATES, WE WANT TO TRY TO ADD 1379 01:01:11,920 --> 01:01:13,160 NUCLEOTIDE SUGARS IN SAMPLER 1380 01:01:13,160 --> 01:01:13,600 PACKS. 1381 01:01:13,600 --> 01:01:14,840 AS WELL AS TRANSFER ACE, DO YOU 1382 01:01:14,840 --> 01:01:17,640 WANT TO TRY AN ENTIRE PANEL OF 1383 01:01:17,640 --> 01:01:18,440 TRANSFERAISES, TRYING TO BRING 1384 01:01:18,440 --> 01:01:20,120 DOWN THE COST BY PRINGING UP THE 1385 01:01:20,120 --> 01:01:22,320 PANEL THAT YOU WOULD HAVE ACCESS 1386 01:01:22,320 --> 01:01:22,480 TO. 1387 01:01:22,480 --> 01:01:25,040 SO I'M NOT ABLE TO COMPLETELY 1388 01:01:25,040 --> 01:01:27,320 DISCLOSE WHO WE WORK WITH BUT WE 1389 01:01:27,320 --> 01:01:29,200 DO HAVE ARRANGEMENTS WITH SOME 1390 01:01:29,200 --> 01:01:30,720 PEOPLE WHO ARE SUPPLYING 1391 01:01:30,720 --> 01:01:32,080 CHEMICALS AND OTHER RE18thS. 1392 01:01:32,080 --> 01:01:34,520 WE'RE ATLANTA SO WE CAN ALSO 1393 01:01:34,520 --> 01:01:35,240 WORK WITH BEVERAGE COMPANIES AND 1394 01:01:35,240 --> 01:01:38,040 WE'VE HEARD OF OTHER PEOPLE 1395 01:01:38,040 --> 01:01:39,240 TALKING ABOUT MMOs SINCE WE 1396 01:01:39,240 --> 01:01:40,840 HAD INTERACTIONS THERE AS WELL 1397 01:01:40,840 --> 01:01:42,040 AND SO THAT'S ONLY REALLY 1398 01:01:42,040 --> 01:01:43,360 FEASIBLE WHEN HAVE YOU THAT 1399 01:01:43,360 --> 01:01:47,240 LARGE STARTING MATERIAL, RIGHT? 1400 01:01:47,240 --> 01:01:48,840 YOU DON'T WANT TO BE ABLE TO SAY 1401 01:01:48,840 --> 01:01:50,640 DO YOU SOMETHING AND THEN YOU GO 1402 01:01:50,640 --> 01:01:53,240 TO THEIR WEBSITE AND YOU CAN'T 1403 01:01:53,240 --> 01:01:55,840 FIND THOSE NUCLEOTIDE SUGARS, 1404 01:01:55,840 --> 01:01:58,320 THEN YOU OF COURSE, THE LAST 1405 01:01:58,320 --> 01:01:59,480 SPEAKER WAS SAYING IF THERE ARE 1406 01:01:59,480 --> 01:02:00,320 THINGS THAT YOU ARE INTERESTED 1407 01:02:00,320 --> 01:02:02,840 IN, IF YOU ARE IN AN RO1, OR 1408 01:02:02,840 --> 01:02:04,400 PRODUCING THINGS THAT NEED 1409 01:02:04,400 --> 01:02:06,480 FEEDBACK ON YOUR PROJECT, CAN BE 1410 01:02:06,480 --> 01:02:07,840 USED BY OTHER PEOPLE WORKING IN 1411 01:02:07,840 --> 01:02:09,280 THAT AREA, THERE'S A GREAT 1412 01:02:09,280 --> 01:02:12,840 CHANCE FOR COLLABORATION THERE, 1413 01:02:12,840 --> 01:02:16,560 WRITING SBIR GROUPS AND WORKING 1414 01:02:16,560 --> 01:02:17,400 TOGETHER THAT WAY. 1415 01:02:17,400 --> 01:02:20,360 SO AFTER LUNCH, WE WILL HAVE 1416 01:02:20,360 --> 01:02:21,440 FREE SAMPLES OF HEALTHY TYPE 1417 01:02:21,440 --> 01:02:24,440 SUGAR, SOMETHING I WOULDN'T HAVE 1418 01:02:24,440 --> 01:02:26,560 SAID PRIOR TO 2017, YOU CAN 1419 01:02:26,560 --> 01:02:29,000 VISIT US AT CHEMILYGLIBBING O 1420 01:02:29,000 --> 01:02:31,080 SCIENCE AND MOVING AND THINKING 1421 01:02:31,080 --> 01:02:32,840 BO THE COMMUNITY, NOT JUST BEING 1422 01:02:32,840 --> 01:02:34,520 THE EXPERTS IN THIS ROOM BUT 1423 01:02:34,520 --> 01:02:36,800 ALSO OTHER BIOLOGISTS AND OTHER 1424 01:02:36,800 --> 01:02:39,360 GLYCO SCIENTISTS WHICH WILL SOON 1425 01:02:39,360 --> 01:02:43,280 BE REBRANDING AT CHEMILY BIO. 1426 01:02:43,280 --> 01:02:50,800 THANKY SO MUCH FOR YOUR 1427 01:02:50,800 --> 01:02:51,240 ATTENTION. 1428 01:02:51,240 --> 01:03:11,440 [ APPLAUSE ] 1429 01:03:11,440 --> 01:03:12,840 EMPLOY STHRKS I GOT A QUESTION 1430 01:03:12,840 --> 01:03:17,160 ABOUT THE AVAILABLE OF UDPIMRKS 1431 01:03:17,160 --> 01:03:18,200 ALNAC, HOPEFULLY THEY'RE 1432 01:03:18,200 --> 01:03:18,840 LISTENING IN TODAY. 1433 01:03:18,840 --> 01:03:26,560 SO OUR NEXT SPEAKER CONTINUES 1434 01:03:26,560 --> 01:03:32,840 THE THEME OF AUTOMATION. 1435 01:03:32,840 --> 01:03:33,360 DR. ALEXEI, DEMCHENKO,. 1436 01:03:33,360 --> 01:03:35,840 >> THANK YOU VERY MUCH DR. POHL, 1437 01:03:35,840 --> 01:03:38,040 IT'S MY PLEASURE TO BE HERE, I'M 1438 01:03:38,040 --> 01:03:39,600 SPEAKING TODAY ON NPI TEAM THAT 1439 01:03:39,600 --> 01:03:42,000 WAS IN CHARGE OF THIS PROJECT, 1440 01:03:42,000 --> 01:03:46,440 IS THERE AN ECHO SOMEWHERE? 1441 01:03:46,440 --> 01:03:47,360 CONSISTING OF MYSELF, KEITH 1442 01:03:47,360 --> 01:03:52,080 STIEN WHO WAS TYPICAL ATTENDEE 1443 01:03:52,080 --> 01:03:54,200 AT THESE MEETINGS AND CHRISTINA 1444 01:03:54,200 --> 01:03:54,600 [INDISCERNIBLE]. 1445 01:03:54,600 --> 01:03:57,600 THIS WAS THE TITLE OF OUR GRANT. 1446 01:03:57,600 --> 01:04:02,640 MY LAB IS WORKING ON CHEMICAL 1447 01:04:02,640 --> 01:04:05,600 GLYCOSYLATION OLIGO SACCHARIDE 1448 01:04:05,600 --> 01:04:08,240 SYNTHESIS AND AUTOMATION, AND OF 1449 01:04:08,240 --> 01:04:09,760 COURSE CHEMICAL GLYCOSYLATION IS 1450 01:04:09,760 --> 01:04:12,440 A CHALLENGE WHEN WE WORK WITH 1451 01:04:12,440 --> 01:04:13,640 CHEMICAL GRID SERVICES 1452 01:04:13,640 --> 01:04:15,640 GLYCOSYLATION, WE NEED TO 1453 01:04:15,640 --> 01:04:17,600 CONSIDER EVERYTHING, LEAVING 1454 01:04:17,600 --> 01:04:18,880 GROUP, REACTING CELLS AND LIVING 1455 01:04:18,880 --> 01:04:22,400 GROUPS GIVE US TROUBLES AS 1456 01:04:22,400 --> 01:04:23,800 DR. WILS ORNAMENT YESTERDAY 1457 01:04:23,800 --> 01:04:24,720 NICEEE ILLUSTRATED WHAT 1458 01:04:24,720 --> 01:04:28,840 PROTECTION GROUPS CAN DO FOR US 1459 01:04:28,840 --> 01:04:34,600 AND SHE INTRODUCED 6 ORTHOGONAL 1460 01:04:34,600 --> 01:04:36,040 PROTECTING GROUPS. 1461 01:04:36,040 --> 01:04:38,520 'S ACTION ITSELF INVOLVES 1462 01:04:38,520 --> 01:04:40,520 PHYSICAL ORGANIC CHEMISTS ARE 1463 01:04:40,520 --> 01:04:42,040 ARGUING WHICH IS THE GOOD, BAD, 1464 01:04:42,040 --> 01:04:45,520 AND UGLY AND EVERYONE KNOWS THAT 1465 01:04:45,520 --> 01:04:46,200 UNCONTROLLED [INDISCERNIBLE] CAN 1466 01:04:46,200 --> 01:04:50,040 LEAD TO A MIX FUR OF ALPHABETTA, 1467 01:04:50,040 --> 01:04:52,640 AS WELL AS OTHER BYPRODUCTS THAT 1468 01:04:52,640 --> 01:04:53,640 BEEN PREDULTED FROM SIDE 1469 01:04:53,640 --> 01:04:56,160 PROCESS, IN MANY WAYS, TPHREU 1470 01:04:56,160 --> 01:04:59,080 RACING COSALATION, THE SINGLE 1471 01:04:59,080 --> 01:04:59,920 STEP GLYCOSYLATION WILL 1472 01:04:59,920 --> 01:05:01,680 CHALLENGE THOSE PEOPLE WORKING 1473 01:05:01,680 --> 01:05:04,440 WITH GLYCANS IN THE SPACE, AND 1474 01:05:04,440 --> 01:05:07,520 STILLFUL [INDISCERNIBLE] OF 1475 01:05:07,520 --> 01:05:11,640 WHOLE ASPECTS OF SYNTHESIS. 1476 01:05:11,640 --> 01:05:12,880 THERE ARE 2 GENERAL WAYS OF 1477 01:05:12,880 --> 01:05:14,240 MAKING TPHREU RACING KAN--KANAS 1478 01:05:14,240 --> 01:05:15,040 THESE DAYS. 1479 01:05:15,040 --> 01:05:19,000 MANUEL SIPGHT SIS THAT HAS MANY 1480 01:05:19,000 --> 01:05:21,400 ADVANTAGES IN TERMS OF STRATEGY 1481 01:05:21,400 --> 01:05:22,160 AND AVAILABLE, AND KNOW HOW 1482 01:05:22,160 --> 01:05:26,520 TECHNIQUES THAT WE,A PLIE AND 1483 01:05:26,520 --> 01:05:27,640 THERE ARE [INDISCERNIBLE] AS 1484 01:05:27,640 --> 01:05:30,400 WELL BECAUSE THEE CAN BE LENGTHY 1485 01:05:30,400 --> 01:05:33,200 AND ENZYMATIC SINCE PEOPLE CAN 1486 01:05:33,200 --> 01:05:35,320 PUBLISH 50 DIFFERENT OLIGIO 1487 01:05:35,320 --> 01:05:36,480 SACCHARIDES AND 1 ARTICLE 1488 01:05:36,480 --> 01:05:38,000 CHEMICAL SINCE PEOPLE PUBLISHED 1489 01:05:38,000 --> 01:05:43,000 1 ARTICLE PER SYNTHESIS. 1490 01:05:43,000 --> 01:05:44,160 SO AUTOMATED APPROACH IN THIS 1491 01:05:44,160 --> 01:05:47,720 RESPECT IS VERY ATTRACTIVE 1492 01:05:47,720 --> 01:05:48,760 BECAUSE COMPLETELY AUTOMATED, 1493 01:05:48,760 --> 01:05:49,960 APPROACH WOULD ALLOW YOU TO WITH 1494 01:05:49,960 --> 01:05:53,880 THE PRESS OF A BUTTON, DO 1495 01:05:53,880 --> 01:05:55,400 SYNTHESIS AND PRODUCE--PRODUCES 1496 01:05:55,400 --> 01:05:57,040 ACCURATELY, THERE ARE 1497 01:05:57,040 --> 01:05:59,600 CHALLENGES, THE CENSUS CAN BE 1498 01:05:59,600 --> 01:06:01,200 INCOMPLETE, IT'S HARDER TO 1499 01:06:01,200 --> 01:06:04,840 CONTROL IN GENERAL, HARDER TO 1500 01:06:04,840 --> 01:06:05,440 MONITOR. 1501 01:06:05,440 --> 01:06:07,280 LARGE BUILDING PLOK ACCESS MAY 1502 01:06:07,280 --> 01:06:09,800 BE NEEDED AND, WELL, ALL 1503 01:06:09,800 --> 01:06:11,320 EQUIPMENT AND EVERYTHING'S BEEN 1504 01:06:11,320 --> 01:06:12,560 DESIGNED FOR PEPTIDE SYNTHESIS 1505 01:06:12,560 --> 01:06:15,960 BUT NOT FOR GLYCAN SYNTHESIS AND 1506 01:06:15,960 --> 01:06:17,000 REGARDLESS THEIR APPROACH, 1507 01:06:17,000 --> 01:06:18,360 BUILDING BLOCKS AND LARGE PART 1508 01:06:18,360 --> 01:06:20,600 ARE MADE STILL MANUELLY, 1509 01:06:20,600 --> 01:06:22,080 ALTHOUGH FIRST AUTOMATION 1510 01:06:22,080 --> 01:06:23,840 EFFORTS HAVE EMERGED AND I 1511 01:06:23,840 --> 01:06:24,880 BELIEVE PROFESSOR PAUL WILL BE 1512 01:06:24,880 --> 01:06:26,840 SPEAKING RIGHT AFTER MY 1513 01:06:26,840 --> 01:06:28,800 PRESENTATION ABOUT AUTOMATION OF 1514 01:06:28,800 --> 01:06:29,640 BUILDING BLOCKS SYNTHESIS AND 1515 01:06:29,640 --> 01:06:52,640 FOR SURE WE WILL HEAR MORE FROM 1516 01:06:52,640 --> 01:06:53,640 CLAIR BENIT TOMORROW. 1517 01:06:53,640 --> 01:06:58,160 --OVER THERE AND PEOPLE IN THIS 1518 01:06:58,160 --> 01:08:52,160 AS WELL, TO STARTING TO DO 1519 01:08:52,160 --> 01:08:53,960 AUTOMATION AS WELL. 1520 01:08:53,960 --> 01:08:59,240 --THAT WOULD ALLOW FOR FULL 1521 01:08:59,240 --> 01:08:59,760 AUTOMATION. 1522 01:08:59,760 --> 01:09:02,640 THANKS TO NIH AND OTHER GRANT, 1523 01:09:02,640 --> 01:09:05,880 OUR LAB PURCHASED NUMBER OF 1524 01:09:05,880 --> 01:09:07,320 STATE-OF-THE-ART ADHAVE YOU VENT 1525 01:09:07,320 --> 01:09:08,880 SYSTEMS THATTA'S USED FOR 1526 01:09:08,880 --> 01:09:11,480 AUTOMATION THESE DAYS AND 1527 01:09:11,480 --> 01:09:14,120 GENERATION B THAT WOULD 1528 01:09:14,120 --> 01:09:16,240 DEVELOPED SHORTLY THEREAFTER, 1529 01:09:16,240 --> 01:09:17,600 INTRODUCED THE FOREIGN CHANGES 1530 01:09:17,600 --> 01:09:19,840 YOU SEE IN RED HERE, WE STARTED 1531 01:09:19,840 --> 01:09:21,120 USING LONGEST SPACER THAT WAS 1532 01:09:21,120 --> 01:09:23,360 PROVEN TO BE BETTER IN THE 1533 01:09:23,360 --> 01:09:25,200 INDEPENDENT STUDY, WE CHANGED 1534 01:09:25,200 --> 01:09:26,800 THE RESIN TO THE GEL WITHOUT A 1535 01:09:26,800 --> 01:09:29,400 PARTICULAR REASON AT THE TIME 1536 01:09:29,400 --> 01:09:31,040 AND ADJUVANT SYSTEM CAME WITH A 1537 01:09:31,040 --> 01:09:33,920 PUMP THAT GAVE US ADDITIONAL 1538 01:09:33,920 --> 01:09:35,320 CAPABILITIES AND AUTOSAMPLER WAS 1539 01:09:35,320 --> 01:09:36,720 USED FOR THE INSERTION TO THE 1540 01:09:36,720 --> 01:09:39,240 SYSTEM OF THE ACTIVATOR. 1541 01:09:39,240 --> 01:09:44,200 THE DONORS WERE CIRCULATED BUT 1542 01:09:44,200 --> 01:09:45,360 MANY ASPECTS REMAIN MANUEL 1543 01:09:45,360 --> 01:09:47,720 BECAUSE WE HAD TO STILL SWITCH 1544 01:09:47,720 --> 01:09:50,600 MANUELLY BETWEEN THE WASTE AND 1545 01:09:50,600 --> 01:09:51,400 DPLI KAN--KANA COLLECTION MODE 1546 01:09:51,400 --> 01:09:54,400 AND ALSO THE BUILDING--THE 1547 01:09:54,400 --> 01:09:56,040 BUILDING PLOKS WERE BOTTLES WERE 1548 01:09:56,040 --> 01:09:57,680 BUILDING BLOCKS WERE SWITCHED 1549 01:09:57,680 --> 01:09:59,480 WITH MANUEL 2, BUT THIS 1550 01:09:59,480 --> 01:10:01,040 GENERATION B WAS AROUND FOR A 1551 01:10:01,040 --> 01:10:03,160 WHILE AND WE USE IT FOR A NUMBER 1552 01:10:03,160 --> 01:10:04,400 OF SYNTHESIS, THIS DPLI 1553 01:10:04,400 --> 01:10:07,200 KAN--KANA STRUCTURE THAT WAS 1554 01:10:07,200 --> 01:10:13,240 ADDITIONAL CHALLENGE FOR US. 1555 01:10:13,240 --> 01:10:14,800 >> FINALLY WE FIGURE OUT TO YOU 1556 01:10:14,800 --> 01:10:18,240 DO IT WITHOUT AUTOMATION, WE 1557 01:10:18,240 --> 01:10:19,080 PUBLISHED PAPERS DESCROIBING OUR 1558 01:10:19,080 --> 01:10:21,720 EFFORTS IN IN RESPECT AND 1559 01:10:21,720 --> 01:10:22,720 COMPLETE AUTOMATION CAPABILITY 1560 01:10:22,720 --> 01:10:24,640 CAME WITH 2 WAY SLIT VALVE THAT 1561 01:10:24,640 --> 01:10:26,920 WOULD ALLOW US TO PROGRAM THE 1562 01:10:26,920 --> 01:10:28,960 MACHINE TO DIRECT THE FLOW 1563 01:10:28,960 --> 01:10:30,880 BETWEEN THE WASTE AND PRODUCT 1564 01:10:30,880 --> 01:10:31,680 COLLECTION MODE. 1565 01:10:31,680 --> 01:10:34,080 AND THE AUTOSAMPLER WAS REPLACED 1566 01:10:34,080 --> 01:10:35,760 WITH THE PREPRATIVE SAMPLER THAT 1567 01:10:35,760 --> 01:10:38,040 ALLOWED US TO INTRODUCE ALSO ALL 1568 01:10:38,040 --> 01:10:39,080 DOLLARS AND ALL REAGENTS 1569 01:10:39,080 --> 01:10:42,320 NECESSARY FOR ALL STEPS OF THE 1570 01:10:42,320 --> 01:10:42,600 SYNTHESIS. 1571 01:10:42,600 --> 01:10:45,080 THAT ALLOWED US TO REPURPOSE OUR 1572 01:10:45,080 --> 01:10:47,160 INTAKE PUMP LINES TO DELIVER 1573 01:10:47,160 --> 01:10:48,440 PURE SOLVENTS WHICH IS REALLY 1574 01:10:48,440 --> 01:10:51,240 IMPORTANT BECAUSE CAN YOU JUST 1575 01:10:51,240 --> 01:11:02,280 BY ACS GREAT, SOLVENTS AND USE 1576 01:11:02,280 --> 01:11:04,040 THEM AS THIS--33% YIELD WHICH IS 1577 01:11:04,040 --> 01:11:06,560 LOWER THAT BEFORE BY 12 HOURS IS 1578 01:11:06,560 --> 01:11:08,760 BECAUSE MY STUDENT WOULD IT IN A 1579 01:11:08,760 --> 01:11:10,440 WAY THAT IT DOESN'T NEED TO 1580 01:11:10,440 --> 01:11:12,320 TESTIMONY BACK IN OUR TIME, THIS 1581 01:11:12,320 --> 01:11:13,960 WAS USED PREVIOUSLY BECAUSE THIS 1582 01:11:13,960 --> 01:11:17,080 WAS SEEN, PRESS THE BUTTON, GO, 1583 01:11:17,080 --> 01:11:20,480 SLEEP, COME BACK IN THE MORNING 1584 01:11:20,480 --> 01:11:21,960 AND FINDED PRODUCT. 1585 01:11:21,960 --> 01:11:23,840 EMPLOY THE STUDENT DOESN'T HAVE 1586 01:11:23,840 --> 01:11:25,080 THE POSSIBILITY TO MONITOR THIS 1587 01:11:25,080 --> 01:11:28,120 REACTION BECAUSE WE DON'T HAVE 1588 01:11:28,120 --> 01:11:31,240 REMOTE ACCESS, IT'S EXPENSIVE 1589 01:11:31,240 --> 01:11:34,600 THESE DAYS, SO ANYWAY, THEY 1590 01:11:34,600 --> 01:11:39,960 PROGRAM THIS TO DO THE 1591 01:11:39,960 --> 01:11:40,400 GLYCOSYLATION. 1592 01:11:40,400 --> 01:11:41,840 IN ADDITION, AND OBVIOUSLY AT 1593 01:11:41,840 --> 01:11:46,680 THIS POINT WE LOST THE 1594 01:11:46,680 --> 01:11:47,480 CAPABILITY OF RECIRCULATING 1595 01:11:47,480 --> 01:11:49,560 WHICH IS A VERY IMPORTANT 1596 01:11:49,560 --> 01:11:54,080 FEATURE OF OUR TECHNOLOGY, MY 1597 01:11:54,080 --> 01:11:55,720 STUDENTS FELT OTHERWISE HE 1598 01:11:55,720 --> 01:11:57,040 REPLACED THE CIRCULATION OF 1599 01:11:57,040 --> 01:11:59,000 DONORS AND PRACTICES MOTIERS TO 1600 01:11:59,000 --> 01:12:00,800 USE WHEN NEEDED AND ALLOWED US 1601 01:12:00,800 --> 01:12:03,600 TO REPURPOSE THE SYSTEM WITH THE 1602 01:12:03,600 --> 01:12:04,520 USE OF ANALYTICAL AUTOSAMPLE 1603 01:12:04,520 --> 01:12:06,640 SINCE NOW THE ONLY SMALL AMOUNTS 1604 01:12:06,640 --> 01:12:09,120 OF DONOR AND PROMOTER WERE 1605 01:12:09,120 --> 01:12:09,400 INJECTED. 1606 01:12:09,400 --> 01:12:11,080 AND WE ALSO DEVELOPED OR OWN 1607 01:12:11,080 --> 01:12:14,080 RESIN THAT WAS NAMED AFTER THE 1608 01:12:14,080 --> 01:12:14,960 INVENTOR [INDISCERNIBLE] 1609 01:12:14,960 --> 01:13:00,600 GRADUATE STUDENT PICTURED HERE. 1610 01:13:00,600 --> 01:13:04,560 WAS BASED ON--FRIENDLY WINDOWS 1611 01:13:04,560 --> 01:13:06,600 BASED SOFTWARE AND WE CAN ALSO 1612 01:13:06,600 --> 01:13:08,080 MOVE TO TAKE A PEEK THROUGH THE 1613 01:13:08,080 --> 01:13:10,960 WINDOW WITH THE SAMPLES DOING, 1614 01:13:10,960 --> 01:13:12,440 IT'S ACCELERATED WINDOW AND IT 1615 01:13:12,440 --> 01:13:14,320 SHOWS THE DRAW SOME SOLVENTS 1616 01:13:14,320 --> 01:13:16,200 PREPARES MIXTURES AS WELL, AND 1617 01:13:16,200 --> 01:13:19,400 DRAWS AND PREPARES MIXTURES IN 1618 01:13:19,400 --> 01:13:21,600 THE SAMPLES SEAT, AND MIX AND 1619 01:13:21,600 --> 01:13:23,480 THEN INJECT AND REINJECT AND 1620 01:13:23,480 --> 01:13:25,760 BRING THE SYSTEM, IT NOW YOU SEE 1621 01:13:25,760 --> 01:13:28,160 THE DEVELOPMENT OF THE COLORS 1622 01:13:28,160 --> 01:13:29,840 AND THE ACID COMBINATION, NATIVE 1623 01:13:29,840 --> 01:13:30,480 AMERICANS IN THE NORTHERNINENCE 1624 01:13:30,480 --> 01:13:33,360 THERE DELIVERED TO THE SYSTEM, 1625 01:13:33,360 --> 01:13:48,040 AND THEN THE NEEDLE GETS CLEAN. 1626 01:13:48,040 --> 01:13:48,440 SO WHAT ELSE? 1627 01:13:48,440 --> 01:13:49,640 VERY RECENTLY WE SWITCHED TO 1628 01:13:49,640 --> 01:13:50,840 AUTOMATION IS SOLUTION, AND I 1629 01:13:50,840 --> 01:13:54,600 WILL TELL YOU WHY, FOR FIRST 1630 01:13:54,600 --> 01:13:56,200 ATTEMPT, WE REALLY DISCOURAGE 1631 01:13:56,200 --> 01:13:58,200 AND THE FIRST ARE THE 1632 01:13:58,200 --> 01:14:03,040 DIFFERENCES, PROCESS AGAIN, I 1633 01:14:03,040 --> 01:14:07,160 READ SINCE THE AUTOMATION AND 1634 01:14:07,160 --> 01:14:10,000 SOLUTION, AND IN THE COLUMN, WE 1635 01:14:10,000 --> 01:14:13,520 ARE STILL USING OUR 1636 01:14:13,520 --> 01:14:18,080 [INDISCERNIBLE] TO RECIRCULATE 1637 01:14:18,080 --> 01:14:21,920 WHERE WE DELIVER ACCEPTANCE FOR 1638 01:14:21,920 --> 01:14:22,160 SAMPLES. 1639 01:14:22,160 --> 01:14:34,040 SO THE FIRST ATTEMPT WAS VERY 1640 01:14:34,040 --> 01:14:35,840 DISCOURAGING--WITH THE DONOR IS 1641 01:14:35,840 --> 01:14:38,360 FAST AND 93% YIELD, SAMEON 1642 01:14:38,360 --> 01:14:40,560 DITIONS IN THE HBLC FAILED 1643 01:14:40,560 --> 01:14:43,200 MISERABLY AFTER 1 HOUR, WE HAVE 1644 01:14:43,200 --> 01:14:47,920 A 25% PRODUCT AND WE KNOW THE 1645 01:14:47,920 --> 01:14:49,360 DOCTOR'S ARTICLE THAT CAN YOU 1646 01:14:49,360 --> 01:14:50,520 FROM MANUEL TOA THE OTHER 1647 01:14:50,520 --> 01:14:51,600 APPROACHES MAY NOT BE SO 1648 01:14:51,600 --> 01:14:52,800 STRAIGHT FORWARD AND WE 1649 01:14:52,800 --> 01:14:54,920 STRUGGLED A LITTLE BIT UNTIL WE 1650 01:14:54,920 --> 01:14:56,160 WANT TO DO THE PLOTS ON THE 1651 01:14:56,160 --> 01:14:58,120 TABLE AND WE WILL LEARN HOW TO 1652 01:14:58,120 --> 01:15:03,240 MAKE IT AS EFFICIENT AS 1653 01:15:03,240 --> 01:15:04,840 [INDISCERNIBLE], ALSO ENGAGED 1654 01:15:04,840 --> 01:15:06,880 HIGH SCHOOL STUDENTS WHO 1655 01:15:06,880 --> 01:15:08,240 PRODUCED THE REACTION WITH THE 1656 01:15:08,240 --> 01:15:09,760 [INDISCERNIBLE] SO THE STUDENT 1657 01:15:09,760 --> 01:15:12,920 WHO WAS GRADUATING NOW SLEEZE IN 1658 01:15:12,920 --> 01:15:17,120 [INDISCERNIBLE]'S LAB, SHE DID 1659 01:15:17,120 --> 01:15:19,320 88% YIELD GLYCOSYLATION, 1660 01:15:19,320 --> 01:15:20,840 UNSTRAINED OFMORE HIGH COOL 1661 01:15:20,840 --> 01:15:24,640 REACTION AND PRODUCED WITH 84, 1662 01:15:24,640 --> 01:15:25,440 86 AND 86% YIELD. 1663 01:15:25,440 --> 01:15:28,240 SO IT'S NOT REALLY AN ONERATED 1664 01:15:28,240 --> 01:15:29,720 DEPENDENT PROCEDURE ONCE IT'S 1665 01:15:29,720 --> 01:15:30,280 PROBABLY REFINED. 1666 01:15:30,280 --> 01:15:33,520 SO NOW WHY ARE WE SO INTERESTED 1667 01:15:33,520 --> 01:15:34,920 IN SOLUTIONS SUDDENLY, OF COURSE 1668 01:15:34,920 --> 01:15:41,040 NOT GIVING UP ON THE POLYMER 1669 01:15:41,040 --> 01:15:42,520 SUPPORTED SYNTHESIS BUT SOLUTION 1670 01:15:42,520 --> 01:15:43,880 BASED APPROACHES WERE DEVELOPED 1671 01:15:43,880 --> 01:15:45,600 DURING THE PANDEMIC BECAUSE THE 1672 01:15:45,600 --> 01:15:47,800 WAY WE DO RESEARCH CHANGED 1673 01:15:47,800 --> 01:15:49,920 SOMEWHAT, OF COURSE WE'RE 1674 01:15:49,920 --> 01:15:51,400 TOTALLY UNPREPARED, WE PANICKED 1675 01:15:51,400 --> 01:15:53,200 BUT EVEN WHEN WE CAME BACK TO 1676 01:15:53,200 --> 01:15:55,960 LABS WE STARTED OPERATING WITH 1677 01:15:55,960 --> 01:15:57,240 REDUCED HOURS, TIME SHARE ALSO, 1678 01:15:57,240 --> 01:16:00,480 IT WAS IMPORTANT FOR US TO LOOK 1679 01:16:00,480 --> 01:16:02,800 AT BATCH SYNTHESIS WITH A PRESS 1680 01:16:02,800 --> 01:16:04,640 OF A BUTTON WE CAN MULTIPLE 1681 01:16:04,640 --> 01:16:06,120 POLARIZED KIEWLS, SET THE 1682 01:16:06,120 --> 01:16:08,400 MACHINE, LEAVE, LET OTHERS IN 1683 01:16:08,400 --> 01:16:11,800 AND THE STUDENT WOULD COME BACK, 1684 01:16:11,800 --> 01:16:14,840 THEY WILL GET MULTIPLE COMPOUNDS 1685 01:16:14,840 --> 01:16:16,640 SYNTHESIZED IN THE ABSENCE. 1686 01:16:16,640 --> 01:16:18,440 SO IN THIS EXAMPLE, OVER HERE WE 1687 01:16:18,440 --> 01:16:21,040 HAVE ONLY 1 DONOR FOR ACCEPTORS 1688 01:16:21,040 --> 01:16:22,680 THAT COULD ALL BE INGESTED AND 1689 01:16:22,680 --> 01:16:26,120 IN A MATTER OF I THINK IN THIS 1690 01:16:26,120 --> 01:16:28,240 CASE, ABOUT 8 HOURS, THE STUDENT 1691 01:16:28,240 --> 01:16:30,760 WILL COME BACK AND FIND FRACTION 1692 01:16:30,760 --> 01:16:33,040 COLLECTION ALL THE PRODUCTS 1693 01:16:33,040 --> 01:16:34,600 SYNTHESIZED, YIELDS ARE NOT 1694 01:16:34,600 --> 01:16:38,120 STELLAR BUT DON'T FORGET THAT'S 1695 01:16:38,120 --> 01:16:39,400 WHERE WE'RE TALKING ABOUT 1696 01:16:39,400 --> 01:16:40,240 COMPLETE AUTOMATION HERE AND TO 1697 01:16:40,240 --> 01:16:42,680 BE HONEST SOME OF THESE YIELDS 1698 01:16:42,680 --> 01:16:45,240 ARE OKAY IN MOST PARTS. 1699 01:16:45,240 --> 01:16:48,080 OTHER EXAMPLE, WHEN THE STUDENT 1700 01:16:48,080 --> 01:16:49,400 RANDOMLY PROGRAMMED A 1701 01:16:49,400 --> 01:16:53,760 COMBINATIONS OF DONORS AND 1702 01:16:53,760 --> 01:16:55,160 ACCEPTORS, THEY ARE ALL LOADED A 1703 01:16:55,160 --> 01:16:59,400 FEW OF EACH AND BY RANDOMLY 1704 01:16:59,400 --> 01:17:00,680 PROGRAMMING COMBINATIONS, THE 1705 01:17:00,680 --> 01:17:02,720 NEXT MORNING AFTER 12 HOUR 1706 01:17:02,720 --> 01:17:04,840 SEQUENCE MY OUTPATIENT SETTINGS 1707 01:17:04,840 --> 01:17:05,880 [INDISCERNIBLE] SHE FOUND 6 1708 01:17:05,880 --> 01:17:07,520 DIFFERENT DISCIPLINARY 1709 01:17:07,520 --> 01:17:17,600 SACCHARIDES AND TRI SACCHARIDES 1710 01:17:17,600 --> 01:17:18,960 IN THE COLLECTORS. 1711 01:17:18,960 --> 01:17:22,600 AS DR. POHL MENTIONED, WE ALSO 1712 01:17:22,600 --> 01:17:23,480 SYNTHESIZED STRUCTURES IN THE 1713 01:17:23,480 --> 01:17:24,480 SOLUTION BASEDDA, PROACH THAT 1714 01:17:24,480 --> 01:17:27,440 ARE NOT PICTURED HERE, WE DID 1715 01:17:27,440 --> 01:17:29,280 THIS AS WELL, THIS WAS NOT 1716 01:17:29,280 --> 01:17:30,360 EXTREMELY EFFICIENT AND THIS IS 1717 01:17:30,360 --> 01:17:32,720 1 OF THE FOCUSES IN OUR LAB 1718 01:17:32,720 --> 01:17:37,240 THESE DAYS, CAN CHRISTINA DEMILL 1719 01:17:37,240 --> 01:17:39,520 WE WORKING ON DECORATING THIS 1720 01:17:39,520 --> 01:17:41,960 AND IT'S ALL DONE NIEWN 1721 01:17:41,960 --> 01:17:42,240 CHEMICALLY. 1722 01:17:42,240 --> 01:17:44,160 THIS PROJECT RESULT INDEED 28 1723 01:17:44,160 --> 01:17:45,240 PUBLICATIONS AND THE MOST RECENT 1724 01:17:45,240 --> 01:17:48,680 1 WAS THANKS TO JOE BARKY WHO 1725 01:17:48,680 --> 01:17:49,600 INVITEDITOUS SCRIEWBT TO THE 1726 01:17:49,600 --> 01:17:51,120 NICE COLLECTION OF GLYCO SCIENCE 1727 01:17:51,120 --> 01:17:54,440 ARTICLES AND THIS DESCRIBES OUR 1728 01:17:54,440 --> 01:17:56,440 GENERATION A, B, & C, 1729 01:17:56,440 --> 01:17:57,640 GENERATION D IS IN THE WORKS AND 1730 01:17:57,640 --> 01:17:59,080 I LOOK FOR MORE DEVELOPMENTS IN 1731 01:17:59,080 --> 01:17:59,440 THIS DEAL. 1732 01:17:59,440 --> 01:18:03,160 SO WHAT CAN YOU DO WITH OUR 1733 01:18:03,160 --> 01:18:04,640 TECHNOLOGY, WE DON'T SELL 1734 01:18:04,640 --> 01:18:06,280 ANYTHING BUT IT'S ALL FREELYY 1735 01:18:06,280 --> 01:18:06,600 AVAILABLE. 1736 01:18:06,600 --> 01:18:11,840 SO YOU ARE WELCOME TO READ OUR 1737 01:18:11,840 --> 01:18:13,120 PAPERS AND [INDISCERNIBLE]. 1738 01:18:13,120 --> 01:18:18,440 TO FINISH THEN, SINCE MICROPHONE 1739 01:18:18,440 --> 01:18:19,120 FELL. 1740 01:18:19,120 --> 01:18:19,600 [LAUGHTER] 1741 01:18:19,600 --> 01:18:21,000 WE COULDN'T HOAFORT MANY 1742 01:18:21,000 --> 01:18:22,680 STUDENTS THESE DAYS BUT WE HAVE 1743 01:18:22,680 --> 01:18:24,320 1 FROM ITALY WHO WILL BRING 1744 01:18:24,320 --> 01:18:27,840 THEIR TECHNOLOGY TO THE OLD 1745 01:18:27,840 --> 01:18:30,240 WORLD SO ANYWAY, IF YOU HAVE 1746 01:18:30,240 --> 01:18:32,800 HPLC AND MANY OF YOU DO WAYED ON 1747 01:18:32,800 --> 01:18:34,040 YOUR PRESENTATIONS CAN YOU DO 1748 01:18:34,040 --> 01:18:37,760 THE SYNTHESIS, CAN YOU REPURPOSE 1749 01:18:37,760 --> 01:18:39,560 SYNTHESIS TO SEPARATE IF WANT TO 1750 01:18:39,560 --> 01:18:42,560 ORE CAN YOU BYPASS THIS. 1751 01:18:42,560 --> 01:18:44,440 ALL RIGHT, CAN THAT, THAT'S ALL 1752 01:18:44,440 --> 01:18:51,920 I HAVE TO DO, IS TO THANK COMMON 1753 01:18:51,920 --> 01:18:54,480 GROUND, TEAM, THIS IS A PHOTY, A 1754 01:18:54,480 --> 01:18:55,440 DIFFERENT PHOTO EVERYONE ELSE IS 1755 01:18:55,440 --> 01:18:57,000 SHOWING SO I PULLED IT FROM MY 1756 01:18:57,000 --> 01:18:58,280 FIRST MEETING WHICH WAS A SECOND 1757 01:18:58,280 --> 01:19:00,120 YEAR OF COMMON FUNDS BECAUSE MY 1758 01:19:00,120 --> 01:19:06,840 COLLABORATORS AND MY GROUP, 1759 01:19:06,840 --> 01:19:59,600 THANK YOU SO MUCH. 1760 01:19:59,600 --> 01:20:03,600 [ APPLAUSE ] 1761 01:20:03,600 --> 01:20:05,240 >> WHERE IS AUTOMATION MOST 1762 01:20:05,240 --> 01:20:05,480 VALUABLE? 1763 01:20:05,480 --> 01:20:08,120 IS IT IMET TORE HAVE SINGLE STEP 1764 01:20:08,120 --> 01:20:10,600 AUTOMATIONS LIKE HE JUST SHOWED 1765 01:20:10,600 --> 01:20:12,440 NEAR THE END OF HIS TALK WHERE 1766 01:20:12,440 --> 01:20:14,480 ALL THE TPHREU RACING 1767 01:20:14,480 --> 01:20:15,040 COSALATIONS ARE AUTOMATED 1768 01:20:15,040 --> 01:20:16,440 BECAUSE THEN HAVE YOU THE ACCESS 1769 01:20:16,440 --> 01:20:18,040 TO THE DISCIPLINARY SACCHARIDE 1770 01:20:18,040 --> 01:20:19,320 OR PENTAWHATEVER IT MIGHT BE SO 1771 01:20:19,320 --> 01:20:21,720 CAN YOU PUT IT INTO THE NEXT 1772 01:20:21,720 --> 01:20:23,200 LIBRARY STEP. 1773 01:20:23,200 --> 01:20:25,040 IT SEEMS LIKE ALEX SESTARTING TO 1774 01:20:25,040 --> 01:20:26,000 GO TOWARD THE SECOND 1 WHERE 1775 01:20:26,000 --> 01:20:29,000 THERE'S A LOT OF VALUE IN 1776 01:20:29,000 --> 01:20:29,600 INDIVIDUALLY AUTOMATING EACH 1777 01:20:29,600 --> 01:20:30,800 STEP AND I HAVE TO SAY THAT'S 1778 01:20:30,800 --> 01:20:32,520 WHERE MY OWN LAB IS ALSO 1779 01:20:32,520 --> 01:20:34,160 STARTING TO WEIGH IN BECAUSE IN 1780 01:20:34,160 --> 01:20:35,960 TERMS OF BROAD UTILITY, THERE'S 1781 01:20:35,960 --> 01:20:38,840 SO MUCH DIVERSITY IN GLYCANS, BY 1782 01:20:38,840 --> 01:20:41,400 ONLY MAKING A SINGLE 1783 01:20:41,400 --> 01:20:42,920 PENTA-WHATEVER SACCHARISE AS 1784 01:20:42,920 --> 01:20:45,280 OPPOSE TO AUTOMATING EACH OF THE 1785 01:20:45,280 --> 01:20:46,120 INDIVIDUAL STEPS WE LOSE 1786 01:20:46,120 --> 01:20:47,160 FLEXIBILITY OVER ALL AND THAT'S 1787 01:20:47,160 --> 01:20:48,840 1 OF THE REASONS WHY WE'RE 1788 01:20:48,840 --> 01:20:50,680 STARTING TO LOOK ALSO AT HOW DO 1789 01:20:50,680 --> 01:20:51,680 WE AUTOMATE INDIVIDUAL STEPS. 1790 01:20:51,680 --> 01:20:53,480 SO 1 OF THE THINGS YOU MAY NOT 1791 01:20:53,480 --> 01:20:56,040 APPRECIATE FOR THOSE OF WHO YOU 1792 01:20:56,040 --> 01:20:57,880 HAVE BEEN THINK THISSING ABOUT 1793 01:20:57,880 --> 01:20:59,760 AUTOMATION FOR DECADES MY DAD 1794 01:20:59,760 --> 01:21:01,440 WAS A TEXTILE WORKER THERE WAS 1 1795 01:21:01,440 --> 01:21:03,240 OF THE FIRST INDUSTRIES THAT WAS 1796 01:21:03,240 --> 01:21:04,600 AUTOMATED HUNDREDS OF YEARS AGO 1797 01:21:04,600 --> 01:21:05,960 SO I ALWAYS GREW UPON WITH THE 1798 01:21:05,960 --> 01:21:08,640 IDEA OKAY, YOU NEED TO DO 1799 01:21:08,640 --> 01:21:11,480 SOMETHINGALUE ADDED AS A HUMAN 1800 01:21:11,480 --> 01:21:15,240 BEING, THE MACHINES CAN DO THE 1801 01:21:15,240 --> 01:21:16,040 REGULAR LABOR. 1802 01:21:16,040 --> 01:21:17,440 WHAT'S AMAZING IS HOW CHEAP THEY 1803 01:21:17,440 --> 01:21:17,960 BECOME. 1804 01:21:17,960 --> 01:21:21,240 YOU WILL SEE HPLC PUMPS OVER AND 1805 01:21:21,240 --> 01:21:22,080 OVER AGAIN, USELESS THE ABILITY 1806 01:21:22,080 --> 01:21:24,400 TO CONTROL THOSE WITH MUCH 1807 01:21:24,400 --> 01:21:26,040 SIMPLER SOFTWARE THAN WHAT WAS 1808 01:21:26,040 --> 01:21:28,360 AVAILABLE 20 YEARS AGO, WHEN I 1809 01:21:28,360 --> 01:21:29,840 STARTED IN 2000, IN MY CAREER 1810 01:21:29,840 --> 01:21:31,440 AND WANTED TO HOME BUILD A 1811 01:21:31,440 --> 01:21:32,920 SYNTHESIZER, I WENT BE OVER TO 1812 01:21:32,920 --> 01:21:33,600 THE ENGINEERING SCHOOL AND 1813 01:21:33,600 --> 01:21:36,080 THEY'RE LIKE THIS IS WAY TOO 1814 01:21:36,080 --> 01:21:37,160 COMPLICATED, THIS IS TOO 1815 01:21:37,160 --> 01:21:38,680 COMPLICATED AND NOW WHAT WE'RE 1816 01:21:38,680 --> 01:21:40,600 SEEING IS THAT THE AUTOMATION 1817 01:21:40,600 --> 01:21:41,400 THAT'S ALREADY AVAILABLE, WE CAN 1818 01:21:41,400 --> 01:21:44,840 MIX AND MATCH THIS, WITH THE 1819 01:21:44,840 --> 01:21:46,920 HPLC PUMPS, PYTHON CODING WHICH 1820 01:21:46,920 --> 01:21:49,840 DIDN'T EXIST 22 YEARS AGO, MAKE 1821 01:21:49,840 --> 01:21:51,360 ITS ALSO EASIER TO HOME BUILD 1822 01:21:51,360 --> 01:21:53,720 THINGS WITHOUT HAVING A LOT OF 1823 01:21:53,720 --> 01:21:54,120 CODING BACKGROUND. 1824 01:21:54,120 --> 01:21:56,040 SO WITH THAT, I WANT TO GIVE YOU 1825 01:21:56,040 --> 01:22:01,000 A LITTLE BIT OF SENSE FOR WHAT 1826 01:22:01,000 --> 01:22:03,040 WE'VE BEEN DOG IN LOOKING AT 1827 01:22:03,040 --> 01:22:03,320 THIS. 1828 01:22:03,320 --> 01:22:05,520 AS I SAID YFULT BUT THE IDEA IS 1829 01:22:05,520 --> 01:22:07,040 TO BE ABLE TO PUSH FORWARD THE 1830 01:22:07,040 --> 01:22:10,600 METHODS FOR BOTH [INDISCERNIBLE] 1831 01:22:10,600 --> 01:22:14,040 ANALITICAL CHEMISTRY AS WELL AS 1832 01:22:14,040 --> 01:22:16,320 ANALYTICAL 60 SIS, THESE ARE ARE 1833 01:22:16,320 --> 01:22:18,440 ALL VERY MUCH INTERLINKED, THESE 1834 01:22:18,440 --> 01:22:19,960 ARE STANDARDS WE MADE THROUGH 1835 01:22:19,960 --> 01:22:23,240 SYNTHESIS, AND WE ISOLATE OR 1836 01:22:23,240 --> 01:22:24,280 NATURAL RESOURCES THROUGH 1837 01:22:24,280 --> 01:22:28,840 ENZYMATIC RESOURCES, YOU DRIVE 1838 01:22:28,840 --> 01:22:29,600 THE ANALYTICAL STANDARDS, CAN 1839 01:22:29,600 --> 01:22:33,160 YOU ALSO RULE OUT STRUCTURES, 1840 01:22:33,160 --> 01:22:33,720 ESPECIALLY ISOMERS BASED ON 1841 01:22:33,720 --> 01:22:35,720 DIFFERENT PROPERTIES THAT ARE 1842 01:22:35,720 --> 01:22:37,440 HARD TO PREDICT DE NOVO BUT IF 1843 01:22:37,440 --> 01:22:40,840 HAVE YOU AUTHENTIC SEASMS YOU 1844 01:22:40,840 --> 01:22:42,880 CAN DEVELOP CHROMATOGRAPHY 1845 01:22:42,880 --> 01:22:46,840 PROTOCOLS OR MASS SPEC THAT CAN 1846 01:22:46,840 --> 01:22:47,720 DIFREBTIATE SPECTERS THAT'S A 1847 01:22:47,720 --> 01:22:49,360 FRONTIER THAT WE'RE OPENING UP 1848 01:22:49,360 --> 01:22:51,760 AS CHEMISTS BECAUSE WE CAN NOW 1849 01:22:51,760 --> 01:22:54,440 MORE EASILY GET TO THE ISOMERS, 1850 01:22:54,440 --> 01:22:55,680 THE ALPHA, BETA, DIFFERENT 1851 01:22:55,680 --> 01:22:58,960 ISOMERS ALL OF WHICH HAVE EXACT 1852 01:22:58,960 --> 01:22:59,800 SAME MOLECULAR WEIGHT IN THE 1853 01:22:59,800 --> 01:23:01,080 MASS SPEC BUT NOW CAN YOU 1854 01:23:01,080 --> 01:23:06,880 DIFFERENTIATE THOSE BY THE 1855 01:23:06,880 --> 01:23:07,320 ISOMERIC RATIOS. 1856 01:23:07,320 --> 01:23:14,880 SO THIS LOOKS LIKE THE WRONG 1857 01:23:14,880 --> 01:23:15,080 TALK. 1858 01:23:15,080 --> 01:23:16,160 THIS LOOKS LIKE YESTERDAY'S 1859 01:23:16,160 --> 01:23:16,360 TALK. 1860 01:23:16,360 --> 01:23:19,320 SO WHAT DO YOU NEED, SO WE'VE 1861 01:23:19,320 --> 01:23:20,160 SEEN MULTILE DIFFERENT 1862 01:23:20,160 --> 01:23:21,360 AUTOMAITIONZ PLATFORMS NOW AND 1863 01:23:21,360 --> 01:23:23,480 WHAT BRINGS ALL THESE TOGETHER 1864 01:23:23,480 --> 01:23:28,000 IS THE ABILITY TO DECIPHER A 1865 01:23:28,000 --> 01:23:30,040 LIQUID HANDLING, A LIQUID 1866 01:23:30,040 --> 01:23:31,840 HANDLING SYSTEM, YOU CAN DELIVER 1867 01:23:31,840 --> 01:23:34,200 SLURYS AND IT CERTAINLY IS DONE 1868 01:23:34,200 --> 01:23:36,920 INDUSTRIAL BUT IT IS CHALLENGING 1869 01:23:36,920 --> 01:23:38,600 THROUGH THE SIZE OF THE SLURY 1870 01:23:38,600 --> 01:23:39,440 ARTICLES CAN ANOTHER VERYIABLE 1871 01:23:39,440 --> 01:23:41,400 YOU HAVE TO CONTROL, SO REALLY 1872 01:23:41,400 --> 01:23:44,600 THE IDEAL PART IS TO BE ABLE TO 1873 01:23:44,600 --> 01:23:45,800 AND TO PUT EVERYTHING INTO 1874 01:23:45,800 --> 01:23:47,920 SOLUTION, YOU HAVE MULTIPLE WAYS 1875 01:23:47,920 --> 01:23:49,160 OF HAVING AUTOMATED DELIVERY 1876 01:23:49,160 --> 01:23:50,760 WHETHER THAT'S THROUGH SERIES 1877 01:23:50,760 --> 01:23:51,960 POINTING PUMPS FOR SMALLER SCALE 1878 01:23:51,960 --> 01:23:57,160 OR THROUGH THE HPLC PUMPS THAT 1879 01:23:57,160 --> 01:23:59,000 WAS JUST SHOWN IN THE PREVIOUS 1880 01:23:59,000 --> 01:23:59,840 PRESENTATION, YOU THERE ARE MANY 1881 01:23:59,840 --> 01:24:02,200 WAYS CAN YOU GET TO THE LIQUID 1882 01:24:02,200 --> 01:24:05,080 HANDLING BUT THEN YOU THINK 1883 01:24:05,080 --> 01:24:06,800 ABOUT THE CHEMISTRY ITSELF, IF 1884 01:24:06,800 --> 01:24:08,120 YOU WANT TO DO SOMETHING WITH 1885 01:24:08,120 --> 01:24:09,760 THE PUMP, HOW DO I DO THAT WITH 1886 01:24:09,760 --> 01:24:11,280 THE CHEMIST RADIOY IN THE 1887 01:24:11,280 --> 01:24:13,600 LITERATURE FOR THE LAST 150 1888 01:24:13,600 --> 01:24:15,280 YEARS OR SO BECAUSE CHEMISTS 1889 01:24:15,280 --> 01:24:17,240 HAVE USED THIS IDEA OF DRIVING A 1890 01:24:17,240 --> 01:24:19,200 REACTION FORWARD, OFTEN TIMES BY 1891 01:24:19,200 --> 01:24:20,440 A PHASE CHANGE WHICH IS EXACTLY 1892 01:24:20,440 --> 01:24:23,360 WHAT DO YOU NOT WANT IN AN 1893 01:24:23,360 --> 01:24:25,120 AUTOMATION PLATFORM, SO A LOT OF 1894 01:24:25,120 --> 01:24:26,440 THE CHALLENGES AS WAS JUST 1895 01:24:26,440 --> 01:24:30,840 ALLUDED TO BEFORE, IS THAT WHEN 1896 01:24:30,840 --> 01:24:32,200 WE DO SOMETHING MANUELLY WE 1897 01:24:32,200 --> 01:24:34,240 DON'T CONTROL A LOT OF HEAT AND 1898 01:24:34,240 --> 01:24:35,680 MASS TRANSFER ISSUES AND WE 1899 01:24:35,680 --> 01:24:37,600 OFTEN TIME VS PURPOSELY DESIGNED 1900 01:24:37,600 --> 01:24:40,160 THE REACTION TO DO A FACE 1901 01:24:40,160 --> 01:24:43,440 TRANSFER CHANGE, THAT PLEENS 1902 01:24:43,440 --> 01:24:45,240 THAT WE CAN'T AUTOMATICALLY JUST 1903 01:24:45,240 --> 01:24:47,440 TRANSFER IT INTO A LIQUID 1904 01:24:47,440 --> 01:24:49,360 HANDLING SYSTEM. 1905 01:24:49,360 --> 01:24:51,200 SO WE STARTED OKAY, THERE WERE 1906 01:24:51,200 --> 01:24:58,040 SOLUTION IN SOLID PHASE METHODS 1907 01:24:58,040 --> 01:25:01,560 OUT THERE TO DO DISP WHAT DO WE 1908 01:25:01,560 --> 01:25:02,040 NEED TO AUTOMATE. 1909 01:25:02,040 --> 01:25:04,040 AS LONG AS YOU HAVE YOUR 1910 01:25:04,040 --> 01:25:06,320 SOLUTION PHASE REACTIONS, YOU 1911 01:25:06,320 --> 01:25:07,040 CAN PROGRAM YOUR 2 BUILDING 1912 01:25:07,040 --> 01:25:08,400 BLOCKS TO COME TOGETHER JUST 1913 01:25:08,400 --> 01:25:12,080 LIKE YOU DO WITH NUCLEIC ACID OR 1914 01:25:12,080 --> 01:25:15,400 PEPTIDEY 60 SIS BUT BRUCE WHO 1915 01:25:15,400 --> 01:25:17,240 INVENTED THIS IDEA BACK IN THE 1916 01:25:17,240 --> 01:25:19,000 1950S AND 60S FOR PEPTIDE 1917 01:25:19,000 --> 01:25:20,600 SYNTHESIS ALSO HAD THE PROBLEM 1918 01:25:20,600 --> 01:25:22,240 OF PEPTIDES WHEN YOU COUPLE THEM 1919 01:25:22,240 --> 01:25:25,320 ONCE THEY GET LONGER IS START 1920 01:25:25,320 --> 01:25:25,760 SELF-AGGREGATING AND 1921 01:25:25,760 --> 01:25:26,560 PRECIPITATING OUT OF SOLUTION, 1922 01:25:26,560 --> 01:25:29,560 SO THIS PROBLEM WAS HE COULD BT 1923 01:25:29,560 --> 01:25:31,400 MAINTAIN SOLUBLE OF HIS PEPTIDES 1924 01:25:31,400 --> 01:25:32,640 AS THEY GROW BUT AS MANY EVER 1925 01:25:32,640 --> 01:25:34,160 YOU KNOW ONCE YOU PROTECT A 1926 01:25:34,160 --> 01:25:37,800 SUGAR YOU DON'T HAVE THE SAME 1927 01:25:37,800 --> 01:25:40,800 PORT OF PROBLEMS IN BASICALLY 1928 01:25:40,800 --> 01:25:43,400 HAVING YOUR SOLUTIONS, YOUR 1929 01:25:43,400 --> 01:25:44,840 SUGARS THEMSELVES PRECIPITATING 1930 01:25:44,840 --> 01:25:48,200 OUT OF SOLUTIONS, IT'S USUALLY 1931 01:25:48,200 --> 01:25:49,440 REAGENTS THAT PRECIPITATE OUT OF 1932 01:25:49,440 --> 01:25:50,880 VARIOUS WAYS. 1933 01:25:50,880 --> 01:25:52,360 SO THOUGHT WE DON'T NEED TO USE 1934 01:25:52,360 --> 01:25:56,240 THE OLD 19 NIFULTS OF WE COULD 1935 01:25:56,240 --> 01:25:57,680 USE SOLUBLE PHASE AND IF WE 1936 01:25:57,680 --> 01:26:02,240 COULD USE TAGS AND WE LOOKED IN 1937 01:26:02,240 --> 01:26:03,440 THE LITERATURE, WE SAW LIPID 1938 01:26:03,440 --> 01:26:04,520 TAGS AND COULD WE DEVELOP A 1939 01:26:04,520 --> 01:26:06,000 SYSTEM THERE, IF YOU WANT TO DO 1940 01:26:06,000 --> 01:26:07,040 A SINGLE REACTION, AS THEY 1941 01:26:07,040 --> 01:26:08,680 SHOWED TOWARD THE END, YOU DON'T 1942 01:26:08,680 --> 01:26:11,480 NEED TAGS AT ALL AND IN FACT 1943 01:26:11,480 --> 01:26:13,760 WE'RE START TO THINK IN WITH 1944 01:26:13,760 --> 01:26:15,040 THE'M PROVEMENTS IN 1945 01:26:15,040 --> 01:26:16,200 CHROMATOGRAPHY THE LAST 20 YEARS 1946 01:26:16,200 --> 01:26:19,600 WE MIGHT NEED TO RETHINK OUR 1947 01:26:19,600 --> 01:26:21,120 FLUORIST TAGS, DO WE NEED THEM 1948 01:26:21,120 --> 01:26:22,560 AS WE SAW WITH OUR 1949 01:26:22,560 --> 01:26:25,160 COLLABORATION, THERE'S A LOT OF 1950 01:26:25,160 --> 01:26:27,480 VALUE IN STILL USING THAT TAG 1951 01:26:27,480 --> 01:26:30,240 FOR THINGS LIKE SULFATED 1952 01:26:30,240 --> 01:26:31,640 COMPOUNDS WHERE IT'S EASIER TO 1953 01:26:31,640 --> 01:26:33,400 FESH OUT. 1954 01:26:33,400 --> 01:26:35,240 SULFATE COMPINEDS ARE DIFFICULT 1955 01:26:35,240 --> 01:26:37,960 TO PURIFY USING STANDARD HPLC 1956 01:26:37,960 --> 01:26:38,440 METHODS. 1957 01:26:38,440 --> 01:26:56,120 THE PROBLEM WITH THESE TAGS OUT 1958 01:26:56,120 --> 01:26:56,440 THERE. 1959 01:26:56,440 --> 01:26:58,040 SO A RIP EDUCATIONAL TAG, YOU 1960 01:26:58,040 --> 01:26:59,360 WOULD SEE, AND THEN YOU TRY TO 1961 01:26:59,360 --> 01:27:01,920 PICK OUT THINGS IN THE NOISE 1962 01:27:01,920 --> 01:27:03,240 PRACTICALLY WITH YOUR SUGAR, SO 1963 01:27:03,240 --> 01:27:04,840 WE SAID LIPID TAGS AREN'T THAT 1964 01:27:04,840 --> 01:27:06,640 USEFUL IF WE WANT TO MONITOR 1965 01:27:06,640 --> 01:27:09,240 REACTIONS AS WE GO ALONG, SO 1966 01:27:09,240 --> 01:27:10,880 WHAT ABOUT POLYETHYLENE TAGS 1967 01:27:10,880 --> 01:27:12,280 THEY HAD BEEN USED WITH THE 1968 01:27:12,280 --> 01:27:13,840 CHEMISTRY DEVELOP THE IN THE 1969 01:27:13,840 --> 01:27:15,720 1990S AND THEY'RE BASICALLY 2 1970 01:27:15,720 --> 01:27:16,880 CARBONS LINK WIDE OXYGEN. 1971 01:27:16,880 --> 01:27:19,920 AND THOSE OF YOU ARE USED TO 1972 01:27:19,920 --> 01:27:21,880 LOOKING AT NMRs, KNOW THAT 1973 01:27:21,880 --> 01:27:23,280 THAT'S EXACTLY LIKE AN OXYGEN 1974 01:27:23,280 --> 01:27:24,920 LINK TO A CARBON AND SUGAR AND 1975 01:27:24,920 --> 01:27:27,280 INDEED YOU LOOK AT THE NMR AND 1976 01:27:27,280 --> 01:27:29,440 THE TAG OVERLAPS EXACTLY WITH 1977 01:27:29,440 --> 01:27:30,040 THE SUGAR. 1978 01:27:30,040 --> 01:27:32,120 SO YOU THOUGHT, WE NEED 1979 01:27:32,120 --> 01:27:32,960 SOMETHING DIFFERENT AND 1980 01:27:32,960 --> 01:27:35,000 [INDISCERNIBLE] HAD BEEN 1981 01:27:35,000 --> 01:27:37,400 STARTING TO USE USE LITTLE 1982 01:27:37,400 --> 01:27:39,040 PIECES OF TEFLON, TAGS THAT THEY 1983 01:27:39,040 --> 01:27:40,720 GET ATTACHED TO ORGANIC 1984 01:27:40,720 --> 01:27:41,800 COMPOUNDS TO MAKE LIBRARIES AND 1985 01:27:41,800 --> 01:27:43,000 WE THOUGHT, YOU KNOW IN THE 1986 01:27:43,000 --> 01:27:45,320 SUGAR CASE, THIS MIGHT BE EASIER 1987 01:27:45,320 --> 01:27:46,240 BECAUSE WOO VERMEN INFECTED A 1988 01:27:46,240 --> 01:27:48,400 LOT OF OXYGENS ON OUR SUGAR THAT 1989 01:27:48,400 --> 01:28:17,240 ALLOW THIS TO DESIRED 1990 01:28:17,240 --> 01:28:18,360 [INDISCERNIBLE]. 1991 01:28:18,360 --> 01:28:20,240 AND THAT WOULD DISCOVER THAT WE 1992 01:28:20,240 --> 01:28:23,880 COULD HAVE LARGE GROWING 1993 01:28:23,880 --> 01:28:25,600 CARBOHYDRATE CHAINS AS WELL AS 1994 01:28:25,600 --> 01:28:28,360 HAVE THE FLUOREST TAG AND STILL 1995 01:28:28,360 --> 01:28:31,200 BE ABLE TO PURIFY OUT OUR 1996 01:28:31,200 --> 01:28:31,440 COMPOUND. 1997 01:28:31,440 --> 01:28:32,840 NOW IF WE WANT TO AUTOMATE THIS, 1998 01:28:32,840 --> 01:28:35,080 OF COURSE THIS HAD TO BE 1999 01:28:35,080 --> 01:28:36,520 PROCESSED WHERE WE HAVE TO 2000 01:28:36,520 --> 01:28:48,720 REINVENT THE YOUR PARTICULAR 2001 01:28:48,720 --> 01:28:49,560 COMPOUND. 2002 01:28:49,560 --> 01:28:51,760 YOU CAN'T AUTOMATE A PROCESS OF 2003 01:28:51,760 --> 01:28:54,040 EVERY SINGLE SUGAR CAN YOU TO DO 2004 01:28:54,040 --> 01:28:55,200 A DIFFERENT PURIFICATION 2005 01:28:55,200 --> 01:28:55,440 PROTOCOL. 2006 01:28:55,440 --> 01:28:56,840 THE OTHER THING WE HAD TO SEE 2007 01:28:56,840 --> 01:28:59,720 WAS WHETHER ASHES TACHING A 2008 01:28:59,720 --> 01:29:01,480 LITTLE C817 TAGGED TO OUR SUGAR 2009 01:29:01,480 --> 01:29:03,880 WOULD ALLOW US TO ACTUALLY BE 2010 01:29:03,880 --> 01:29:09,840 ABLE TO DO THIS KIND OF 2011 01:29:09,840 --> 01:29:13,120 CHEMISTRY AND DO A PURIFICATION 2012 01:29:13,120 --> 01:29:14,120 SEPARATION REGARDLESS OF WHAT 2013 01:29:14,120 --> 01:29:16,760 THE TAG MIGHT NEED. 2014 01:29:16,760 --> 01:29:18,280 AND WE FOUND OUT THAT WE CAN 2015 01:29:18,280 --> 01:29:20,800 RERNTION GARDLESS OF THE SUGAROT 2016 01:29:20,800 --> 01:29:21,880 FLUORIST TAG AND NOT OHM THAT 2017 01:29:21,880 --> 01:29:24,800 ONCE WE LOOK ATOTMATION 2018 01:29:24,800 --> 01:29:26,040 PLATFORMS, LOOKING IN AT CHEM 2019 01:29:26,040 --> 01:29:27,840 SPEED THAT WE MACHINE SHOP TO 2020 01:29:27,840 --> 01:29:31,080 MODIFY, IT TURNS OUT BECAUSE 2021 01:29:31,080 --> 01:29:33,520 THAT FLOOR IS GEL THAT THE FIRST 2022 01:29:33,520 --> 01:29:36,640 TACK ALLOWS TO YOU DO THE 2023 01:29:36,640 --> 01:29:37,880 SEPARATION DOESN'T STICK TO 2024 01:29:37,880 --> 01:29:40,120 OTHER COMPOUNDS THE WAY OUR 2025 01:29:40,120 --> 01:29:43,120 NORMAL 1 DOES, AND REACH THAT 2026 01:29:43,120 --> 01:29:45,600 SAME OVER AND OVER AGAIN, SO 1 2027 01:29:45,600 --> 01:29:46,520 THING MANY DON'T APPRECIATE IS 2028 01:29:46,520 --> 01:29:49,040 THE REASON WHY WE COULD AUTOMATE 2029 01:29:49,040 --> 01:29:56,560 THIS PROCESS IS BECAUSE THE THIS 2030 01:29:56,560 --> 01:29:58,040 RINSES SO CLEAN THAT WE COULD 2031 01:29:58,040 --> 01:29:59,600 GET AWAY WITH THE PROCESS. 2032 01:29:59,600 --> 01:30:00,640 SO YOU SEE THE MACHINE WITH 2033 01:30:00,640 --> 01:30:02,040 THIS, WE HAVE THE BUILDING PLOKS 2034 01:30:02,040 --> 01:30:04,040 THAT WE NEED TO BE ABLE TO GET 2035 01:30:04,040 --> 01:30:07,320 TO THOSE, WE NEED REAGENTS TO 2036 01:30:07,320 --> 01:30:08,440 COUPLE THE SUGARS THEMSELVES AND 2037 01:30:08,440 --> 01:30:10,360 WE NEED A PURIFICATION SYSTEM 2038 01:30:10,360 --> 01:30:12,120 FOR THIS SORT OF MULTISTEP 2039 01:30:12,120 --> 01:30:12,800 SYNTHESIS. 2040 01:30:12,800 --> 01:30:28,880 SO THIS IS WHAT WE ULTIMATELY 2041 01:30:28,880 --> 01:30:30,440 DEVELOPED THAT IT INHIBITED 2042 01:30:30,440 --> 01:30:32,160 THEIR ABILITY TO MOVE ON TO MAKE 2043 01:30:32,160 --> 01:30:34,640 THE DPLI KAN--KANAS THEMSELVES. 2044 01:30:34,640 --> 01:30:36,440 THIS IS A BIT OF A PICTURE TO 2045 01:30:36,440 --> 01:30:38,360 SHORTENED WHAT WE'RE SEEING HERE 2046 01:30:38,360 --> 01:30:39,360 BUT THE ROUGH UPPER LEFT HAND 2047 01:30:39,360 --> 01:30:40,440 CORNER YOU SEE THE NEEDLE ARM 2048 01:30:40,440 --> 01:30:42,320 AND HAD IS THE BASIC PROCESS FOR 2049 01:30:42,320 --> 01:30:48,040 ALL THESE DEFINITE AUTOMATION 2050 01:30:48,040 --> 01:30:49,800 PROCESSES THIS IS A NEEDLE ARM 2051 01:30:49,800 --> 01:30:51,640 THAT TAKE THE SOLUTION OUT OF A 2052 01:30:51,640 --> 01:31:05,040 CONTAINER JUST LIKE THE HPLC IF 2053 01:31:05,040 --> 01:31:06,560 YOU HAVE A RESIDENT IN A 2054 01:31:06,560 --> 01:31:09,040 CARTRIDGE, SO WE CAN GO DOWN TO 2055 01:31:09,040 --> 01:31:10,480 MINUS 40-DEGREE FIST WE WANT TO 2056 01:31:10,480 --> 01:31:15,400 DO LOW TEMPERATURE REACTIONS, 2057 01:31:15,400 --> 01:31:17,160 AND WE CAN REFLEX WE WANT TO, 2058 01:31:17,160 --> 01:31:20,240 ALTHOUGH IT'S LESS USEFUL, SO UP 2059 01:31:20,240 --> 01:31:21,720 THERE IS THE EXTRACTION MEDIAN 2060 01:31:21,720 --> 01:31:22,040 FROM IT. 2061 01:31:22,040 --> 01:31:24,920 SO THIS IS THE ACTUAL PROCESS, 2062 01:31:24,920 --> 01:31:26,600 WE TAKE THE COMPOUND AND WE PUT 2063 01:31:26,600 --> 01:31:28,680 THEM TOGETHER AND LOAD IT ON TO 2064 01:31:28,680 --> 01:31:32,320 OUR SILICA GEL AND THEN BY 2065 01:31:32,320 --> 01:31:34,200 TAKING OUR AQUEOUS MIX THUR 2066 01:31:34,200 --> 01:31:37,040 ALLOWS THE INTERACTION FOR THE 2067 01:31:37,040 --> 01:31:39,400 COP BINNATION TO ADHERE TO THE 2068 01:31:39,400 --> 01:31:41,320 COLUMN AND EVERYTHING ALOWTS OUT 2069 01:31:41,320 --> 01:31:44,080 AND THEN WE USE A PURE ORGANIC 2070 01:31:44,080 --> 01:31:45,720 SOLVE TONIGHT ALLUDE OFF OUR 2071 01:31:45,720 --> 01:31:46,480 DESIRED COMPOUND AND SO OF 2072 01:31:46,480 --> 01:31:49,080 COURSE WE ARE DOING THIS FIRST 2073 01:31:49,080 --> 01:31:50,040 WITH NEUTRAL COMPOUNDS. 2074 01:31:50,040 --> 01:31:52,240 AND STARTED LOOKING AT THIS IN 2075 01:31:52,240 --> 01:31:53,640 VARIOUS THINGS. 2076 01:31:53,640 --> 01:31:58,040 HERE'S AN EXAMPLE OF WHAT I WAS 2077 01:31:58,040 --> 01:31:58,600 TALKING ABOUT BEFORE. 2078 01:31:58,600 --> 01:32:09,880 THE FLOR --FLOR, TAG AND IS 2079 01:32:09,880 --> 01:32:10,240 INCORPORATED. 2080 01:32:10,240 --> 01:32:11,880 CAN WE INCORPORATE A SULFATE 2081 01:32:11,880 --> 01:32:13,640 INTO THE PROTOCOLS AND WOULD WE 2082 01:32:13,640 --> 01:32:15,760 BE ABLE TO DO THE SOLID PHASE 2083 01:32:15,760 --> 01:32:18,960 EXTRACTION OR WOULD NOW, THAT 2084 01:32:18,960 --> 01:32:19,960 TEGGIC--STRATEGIC 5LY CHARGED 2085 01:32:19,960 --> 01:32:22,200 SULF 8 GO THROUGH IN THE WATER, 2086 01:32:22,200 --> 01:32:24,920 IT WOULDN'T ADHERE TO THE 2087 01:32:24,920 --> 01:32:26,240 EXTRACTION COLUMN. 2088 01:32:26,240 --> 01:32:28,760 AND VICTORIA AN NIH POST DOC 2089 01:32:28,760 --> 01:32:29,840 FELLOW AT THE UNIVERSITY OF 2090 01:32:29,840 --> 01:32:31,320 UTAH, SHE WAS LOOKING FAVORITE 2091 01:32:31,320 --> 01:32:37,080 CAN WE GET TRANSFER PROCESS AND 2092 01:32:37,080 --> 01:32:39,280 IT URN ITS OUT THE COMMON 2093 01:32:39,280 --> 01:32:47,240 SULFATION PROTOCOLS THAT WE USE 2094 01:32:47,240 --> 01:32:53,320 AT THE KONSZITRATIONS 2095 01:32:53,320 --> 01:32:54,680 THAT--CONCENTRATIONS WHEN YOU 2096 01:32:54,680 --> 01:32:56,440 CAN CAN'TAD AID SOLID INTO YOUR 2097 01:32:56,440 --> 01:32:58,840 REACTION, AND YOU SART OUT WITH 2098 01:32:58,840 --> 01:33:00,160 A SALVATIONARMYUSAENT AND ADD 2099 01:33:00,160 --> 01:33:02,360 SOLUTION AND IT DESOLVES. 2100 01:33:02,360 --> 01:33:04,000 AFTER TRYING MANY THINGS I 2101 01:33:04,000 --> 01:33:06,080 ENCOURAGE HER TO LOOK AT THE 2102 01:33:06,080 --> 01:33:07,240 PAPER PER DETAILS, AND SHE 2103 01:33:07,240 --> 01:33:09,400 DISCOVERED THAT SHE COULD USE TO 2104 01:33:09,400 --> 01:33:11,360 BE ABLE TO DEVELOP THIS 2105 01:33:11,360 --> 01:33:12,960 AUTOMATION PROTOCOL AND IN THIS 2106 01:33:12,960 --> 01:33:16,200 CASE, MAKE THESE SOLVATED RAMINS 2107 01:33:16,200 --> 01:33:18,240 AND YOU COULD HAVE THE SOLVATED 2108 01:33:18,240 --> 01:33:20,280 COMPOUND STICK TO THE PHASE, 2109 01:33:20,280 --> 01:33:22,080 EXTRACTION MEDIA AND DO THIS 2110 01:33:22,080 --> 01:33:22,960 KIND OF PURIFICATION PROTOCOL 2111 01:33:22,960 --> 01:33:24,760 AND THAT WAS THE STARTING POINT 2112 01:33:24,760 --> 01:33:27,040 FOR THE FANTASTIC COLLABORATION 2113 01:33:27,040 --> 01:33:28,160 WITH LINDA WILSON'S LAB THAT YOU 2114 01:33:28,160 --> 01:33:29,520 HEARD ABOUT YESTERDAY IN BEING 2115 01:33:29,520 --> 01:33:32,160 ABLE TO APPLY THIS SORT OF 2116 01:33:32,160 --> 01:33:35,520 CHEMISTRY TO THE GLYCO IMMUNO 2117 01:33:35,520 --> 01:33:35,760 GLYCANS. 2118 01:33:35,760 --> 01:33:38,080 WE ARE ALSO LOOKING AT DEFINITE 2119 01:33:38,080 --> 01:33:40,640 PROTOCOLS AND I WON'T INTO ANY 2120 01:33:40,640 --> 01:33:45,120 DETAILS BUT WE ALSO HAVE BEEN 2121 01:33:45,120 --> 01:33:46,840 WORKING ON INCORPORATING A WIDE 2122 01:33:46,840 --> 01:33:49,040 VARIETY OF CHEMIST RADIOYS INTO 2123 01:33:49,040 --> 01:33:50,120 THIS PROTOCOL AND WE HAVEN'T 2124 01:33:50,120 --> 01:33:52,960 SEEN A LIMIT TO THE SOLID PHASE 2125 01:33:52,960 --> 01:33:54,040 EXTRACTION TAG IN TERMS OF 2126 01:33:54,040 --> 01:33:57,480 CHEMISTRIES THAT WE'RE DOING, IT 2127 01:33:57,480 --> 01:33:58,640 IS ELECTRON WITH DRAWING SO WE 2128 01:33:58,640 --> 01:34:01,120 HAVE A BIT OF A SPACER BETWEEN 2129 01:34:01,120 --> 01:34:03,480 THE CF817 AND OUR SUGAR BECAUSE 2130 01:34:03,480 --> 01:34:04,800 OTHERWISE IF IT'S TOO CLOSE IT 2131 01:34:04,800 --> 01:34:05,680 WILL AFFECT THE CHEM RIEOF THEY 2132 01:34:05,680 --> 01:34:07,640 AS CAN YOU IMAGINE, BUT 2133 01:34:07,640 --> 01:34:10,240 OTHERWISE THIS COULD WORK QUITE 2134 01:34:10,240 --> 01:34:10,480 NICELY. 2135 01:34:10,480 --> 01:34:12,040 WE ALSO AS I MENTIONED YESTERDAY 2136 01:34:12,040 --> 01:34:15,960 HAVE BEEN WORKING ON TRYING TO 2137 01:34:15,960 --> 01:34:16,800 PUT GLYCO [INDISCERNIBLE]. 2138 01:34:16,800 --> 01:34:18,320 OTHER DIFFERENT TYPES OF CHEMIST 2139 01:34:18,320 --> 01:34:20,200 RADIOYY THAT YOU DON'T GET IN 2140 01:34:20,200 --> 01:34:21,280 NATURAL PROWCTD PLODUCTS AND 2141 01:34:21,280 --> 01:34:24,600 WOULD BE IMPOSSIBLE TO DO WITH 2142 01:34:24,600 --> 01:34:25,120 THE CHEMISTRY. 2143 01:34:25,120 --> 01:34:27,080 LIKE I SAID YESTERDAY, WE ARE 2144 01:34:27,080 --> 01:34:29,320 WORKING TOGETHER SO QUEE FOLK 2145 01:34:29,320 --> 01:34:31,600 THE PROBLEM WE CAN DO WITH THE 2146 01:34:31,600 --> 01:34:32,720 NATURAL ISOLATION OR ENZYME 2147 01:34:32,720 --> 01:34:34,240 CHEMISTRY AND CERTAINLY MAKING 2148 01:34:34,240 --> 01:34:36,640 THIS KIND OF SULFUR LINKAGE FOR 2149 01:34:36,640 --> 01:34:37,400 STABILITY AGAINST ENZYMES 2150 01:34:37,400 --> 01:34:39,280 CLEAVAGE IS 1 OF THOSE KINDS OF 2151 01:34:39,280 --> 01:34:40,240 CATEGORIES AND ONCE AGAIN IT 2152 01:34:40,240 --> 01:34:41,240 DEPEND OFFICE OF DIVERSITY BEING 2153 01:34:41,240 --> 01:34:46,200 ABLE TO FIND SLIGHTLY DIFFERENT 2154 01:34:46,200 --> 01:34:47,440 CHEMIST RADIORYS AND PROTOCOLS 2155 01:34:47,440 --> 01:34:50,040 TO GET THIS LINKAGE TO COME IN. 2156 01:34:50,040 --> 01:34:51,840 WE COULD APPLY IT TO OTHER 2157 01:34:51,840 --> 01:34:53,800 DIFFERENT KINDS OF SUGARS. 2158 01:34:53,800 --> 01:34:56,520 SO I SAID YESTERDAY, THIS IS ALL 2159 01:34:56,520 --> 01:34:59,400 GREAT BUT THE REASON WHY WE ARE 2160 01:34:59,400 --> 01:35:01,960 STILL MAKING HUGE AMOUNTS IS THE 2161 01:35:01,960 --> 01:35:04,120 BUILDING BLOCKS STATION AND WHY 2162 01:35:04,120 --> 01:35:05,040 CAN'T WE AUTOMATE THESE? 2163 01:35:05,040 --> 01:35:14,800 WHY IS IT THAT WE'RE DOING ALL 2164 01:35:14,800 --> 01:35:16,280 THESE STEPS MANUELLY THEN WE 2165 01:35:16,280 --> 01:35:17,840 MAKE COMPOUNDS AND THAT'S WHAT 2166 01:35:17,840 --> 01:35:19,000 MAKES THE AUTOMATED BUILDING 2167 01:35:19,000 --> 01:35:21,640 CLOCKS OF DEC NOT JUST THE DPLI 2168 01:35:21,640 --> 01:35:22,760 KAN--KANAS THAT THESE THEMSELVES 2169 01:35:22,760 --> 01:35:24,720 BECAUSE OTHERWISE WE DON'T HAVE 2170 01:35:24,720 --> 01:35:26,600 AUTOMATION DPLI KAN--KANA 2171 01:35:26,600 --> 01:35:26,840 SYNTHESIS. 2172 01:35:26,840 --> 01:35:29,120 AND SO, THIS IS WHERE WE 2173 01:35:29,120 --> 01:35:30,880 TRANSATION, SOME OF YOU ASKED ME 2174 01:35:30,880 --> 01:35:31,840 THIS UNIVERSITY, SO I THOUGHT IT 2175 01:35:31,840 --> 01:35:36,080 WAS WORTH A SLIDE HERE, WE'RE 2176 01:35:36,080 --> 01:35:39,320 USELY DOING THINGS, FOR 2177 01:35:39,320 --> 01:35:40,120 ENSIKEATIC CHEMIST RADIOY, THAT 2178 01:35:40,120 --> 01:35:42,360 MEANS CAN YOU STILL USE LIQUID 2179 01:35:42,360 --> 01:35:47,320 HANDLING TO PUT BOTH YOUR 2180 01:35:47,320 --> 01:35:48,720 COMPOUNDS INTO THE GLASS REACTOR 2181 01:35:48,720 --> 01:35:49,760 BUT THEN YOU'RE LIMITED TO BY 2182 01:35:49,760 --> 01:35:52,480 THE SYSTEM OF THE REACTOR, WE 2183 01:35:52,480 --> 01:35:54,160 COULD DO A PARALLEL REACTION BUT 2184 01:35:54,160 --> 01:35:55,680 THERE'S EMILY THE TO THE SCALE 2185 01:35:55,680 --> 01:35:56,600 BASED ON THE REACTOR. 2186 01:35:56,600 --> 01:36:00,080 BUT IF YOU LOOK AT THE CHEMICAL 2187 01:36:00,080 --> 01:36:00,800 INDUSTRY ESPECIALLY PROCESSING 2188 01:36:00,800 --> 01:36:02,800 NATURAL GAZ AND THE GASOLINE, 2189 01:36:02,800 --> 01:36:05,040 OIL PRODUCTS THEY DO THAT IN 2190 01:36:05,040 --> 01:36:05,240 FLOW. 2191 01:36:05,240 --> 01:36:12,760 THAT THAT YOU BASICALLY TAKE 2192 01:36:12,760 --> 01:36:14,840 YOUR STARTING MATERIALS AND THE 2193 01:36:14,840 --> 01:36:16,520 REACTION HAPPENS IN THE TUBE AND 2194 01:36:16,520 --> 01:36:18,800 IT STATES CONSTANT BECAUSE THE 2195 01:36:18,800 --> 01:36:20,160 REACTION IN THE SAME 2196 01:36:20,160 --> 01:36:21,080 [INDISCERNIBLE] ULTIMATELY. 2197 01:36:21,080 --> 01:36:22,640 NOW YOU HAVE TO RETHINK 2198 01:36:22,640 --> 01:36:24,080 CHEMISTRY BECAUSE NOW YOU WANT 2199 01:36:24,080 --> 01:36:25,680 FAST, YOU DON'T WANT TO HAVE TO 2200 01:36:25,680 --> 01:36:27,800 CHUG THIS ALONG SO SLOWLY IN A 2201 01:36:27,800 --> 01:36:30,000 TUBE THAT YOU'RE RUNNING 2202 01:36:30,000 --> 01:36:32,200 REACTIONS FOR 5, 6, 7 HOURS AND 2203 01:36:32,200 --> 01:36:33,600 FLANKLY ON THE AUTOMATION 2204 01:36:33,600 --> 01:36:37,080 PLATFORM, WE HAVE THE REACTION 2205 01:36:37,080 --> 01:36:38,560 THAT MORPHS FOR POWERS BECAUSE 2206 01:36:38,560 --> 01:36:40,440 THE FLAT FORM IS MONEY AND SO 2207 01:36:40,440 --> 01:36:42,720 LESS TYPE HAVE YOU ON THAT, THE 2208 01:36:42,720 --> 01:36:45,240 BETTER THE PROCESS BECOMES 2209 01:36:45,240 --> 01:36:47,880 OVERALL AND SO, SCALING REACTION 2210 01:36:47,880 --> 01:36:50,800 TO MATCH, HAS A LOT OF PATHWAY 2211 01:36:50,800 --> 01:36:53,600 GIVES RAMSTERS AS WE ORDER FROM 2212 01:36:53,600 --> 01:36:54,760 THE PREVIOUS GROUP, IT'S 2213 01:36:54,760 --> 01:36:55,600 CONTINUOUS FLOW, ONCE HAVE YOU 2214 01:36:55,600 --> 01:36:57,640 THE PROCESS YOU CAN RUN IT FOR A 2215 01:36:57,640 --> 01:36:59,240 PERIOD OF TIME AND THEREBY GET 2216 01:36:59,240 --> 01:37:01,000 MORE PRODUCT, SO YOU DON'T HAVE 2217 01:37:01,000 --> 01:37:02,440 TO DO REOPTIMIZATION, AND THIS 2218 01:37:02,440 --> 01:37:04,240 IS OUR PATHWAY TO THE 15 YEAR 2219 01:37:04,240 --> 01:37:06,320 GOLD TO VERY QUICKLY AND EASILY 2220 01:37:06,320 --> 01:37:07,160 GROW AN INDUSTRY AROUND THIS 2221 01:37:07,160 --> 01:37:12,560 WHICH IS WHAT I WAS EXCITED 2222 01:37:12,560 --> 01:37:13,960 ABOUT HIDO WE GROW THESE AROUND 2223 01:37:13,960 --> 01:37:15,560 THIS TO BE ABLE TO SCALE 2224 01:37:15,560 --> 01:37:18,760 EVERYTHING THAT WE MAKE AND THIS 2225 01:37:18,760 --> 01:37:20,840 IS THE ANAGRAM OF FLOW CAM IS 2226 01:37:20,840 --> 01:37:22,240 THE OPEN SOURCE SOFTWARE TO 2227 01:37:22,240 --> 01:37:23,400 PROGRAM THESE THESE. 2228 01:37:23,400 --> 01:37:24,800 SO WE TALKED ABOUT THIS ALSO 2229 01:37:24,800 --> 01:37:25,840 AFTER WE WORK WIDE CLAY BENIT 2230 01:37:25,840 --> 01:37:27,560 AND YOU WILL HEAR MORE ABOUT HIS 2231 01:37:27,560 --> 01:37:28,000 WORK LATER. 2232 01:37:28,000 --> 01:37:31,000 WE NOW HAVE THESE PROCESSES, 2233 01:37:31,000 --> 01:37:33,800 WHERE WE CAN COMBINE PUMPS, THE 2234 01:37:33,800 --> 01:37:37,240 SYRINGE PUMPS OR HPLC PUMPS, WE 2235 01:37:37,240 --> 01:37:39,800 CAN DO ORTH1 REACTION OR WE CAN 2236 01:37:39,800 --> 01:37:42,440 FIEBD COMMON SELF-ENTS SO WE CAN 2237 01:37:42,440 --> 01:37:43,680 [INDISCERNIBLE] REACTIONS AND 2238 01:37:43,680 --> 01:37:44,840 START MAKING ENTIRE BUILDING 2239 01:37:44,840 --> 01:37:46,040 PLOKS FROM FLOW AND THIS, YOU 2240 01:37:46,040 --> 01:37:47,840 KNOW IT TOOK THEM ABOUT A WEEK 2241 01:37:47,840 --> 01:37:50,080 TO MAKE THIS MANUELLY AND 2242 01:37:50,080 --> 01:37:52,200 BASICALLY A MORNING TO MAKE THIS 2243 01:37:52,200 --> 01:37:53,680 IN A FLOW PROCESS, OBVIOUSLY YOU 2244 01:37:53,680 --> 01:37:54,720 HAVE TO RUN IT LONGER IF YOU 2245 01:37:54,720 --> 01:37:57,600 WANT TO DO IT ON A KILOSCALE BUT 2246 01:37:57,600 --> 01:38:00,720 THE IDEA IS THAT YOU SAVE A LOT 2247 01:38:00,720 --> 01:38:03,520 OF LABOR AND HALF A DAY TO MAKE 2248 01:38:03,520 --> 01:38:05,720 A BUILDING PLOOK AND WHAT I WANT 2249 01:38:05,720 --> 01:38:08,600 BECAUSE IF I HAVE AN IDEA OR 2250 01:38:08,600 --> 01:38:10,600 HAVE YOU AN IDEA, OR WEEKS TO 2251 01:38:10,600 --> 01:38:12,400 MAKE THE BLOCK BEFORE WE GET TO 2252 01:38:12,400 --> 01:38:12,920 THE GLYCAN SYNTHESIS. 2253 01:38:12,920 --> 01:38:15,680 SO THIS IS THE FIRST EXAMPLE OF 2254 01:38:15,680 --> 01:38:17,280 AUTOMATED CARBOHYDRATE BUILDING 2255 01:38:17,280 --> 01:38:18,120 PLOKS SYNTHESIS AND WE'RE 2256 01:38:18,120 --> 01:38:19,440 EXCITED TO KEEP GOING WITH THIS 2257 01:38:19,440 --> 01:38:21,840 PROCESS TO BE ABLE TO AUTOMATE 2258 01:38:21,840 --> 01:38:23,600 ALL SORTS OF DIFFERENT BUILDING 2259 01:38:23,600 --> 01:38:25,520 PLOKS, INCLUDING THE 1S INVOLVED 2260 01:38:25,520 --> 01:38:26,800 IN PEPTIDE O DPLI KAN--KANA 2261 01:38:26,800 --> 01:38:28,200 SYNTHESIS AND THAT'S WHAT WE 2262 01:38:28,200 --> 01:38:31,000 HEAR ABOUT NEXT IS HOW DO WE USE 2263 01:38:31,000 --> 01:38:33,640 SOME OF THESE PROWCTDS THAT GEP 2264 01:38:33,640 --> 01:38:35,520 ANALYTICAL TECH NEEBS BECAUSE WE 2265 01:38:35,520 --> 01:38:36,360 HAVE AUTHENTIC SAMPLES AND 2266 01:38:36,360 --> 01:38:39,360 FINALLY I WANT TO MAKE SURE IT'S 2267 01:38:39,360 --> 01:38:42,720 VERY CLEAR, AUTOMATION REQUIRES 2268 01:38:42,720 --> 01:38:43,760 HUMAN CREATIVITY, THE ICS SIGHT 2269 01:38:43,760 --> 01:38:44,960 8 HOURS THING ABOUT AUTOMATION 2270 01:38:44,960 --> 01:38:48,840 IS YOU CAN GET RID OF A LOT OF 2271 01:38:48,840 --> 01:38:51,800 TEDIOUS WORK, CLONING IS CHEAPER 2272 01:38:51,800 --> 01:38:55,320 NOW THAN BACK IN THE 1700S WHEN 2273 01:38:55,320 --> 01:38:57,760 EVERYTHING HAD BE MADE MANUELLY. 2274 01:38:57,760 --> 01:38:59,080 THAT'S THE SAME PROCESS I SEE 2275 01:38:59,080 --> 01:39:01,600 HERE, WE CAN SHIFT OUR LABOR TO 2276 01:39:01,600 --> 01:39:03,600 TEDEIUM LABOR TO THINK ABOUT 2277 01:39:03,600 --> 01:39:05,000 PROBLEMS AND DEVELOPING 2278 01:39:05,000 --> 01:39:06,280 CHEMISTRY AND QUICKLY DEVELOP 2279 01:39:06,280 --> 01:39:07,560 AND TRANSFERRING THAT TO OTHER 2280 01:39:07,560 --> 01:39:08,120 LABS. 2281 01:39:08,120 --> 01:39:10,680 SO WITH THAT, I THANK YOU FOR MY 2282 01:39:10,680 --> 01:39:12,120 FORGZ OF THIS PORTION AND THEN I 2283 01:39:12,120 --> 01:39:15,360 WOULD LIKE TO TRANSITION TO 2284 01:39:15,360 --> 01:39:16,200 DR. KRISTI PETERSON'S WORK WHO 2285 01:39:16,200 --> 01:39:17,680 WE'VE BEEN WORKING WITH AS PART 2286 01:39:17,680 --> 01:39:19,520 OF A SUPPLEMENT TO LOOK AT 2287 01:39:19,520 --> 01:39:20,320 PEPTIDE DPLI KAN--KANAS AND 2288 01:39:20,320 --> 01:39:23,680 WHERE WE'RE WORKING ON BUILDING 2289 01:39:23,680 --> 01:39:26,080 DPLOK SYNTHESIS PERLET PEPTIDE 2290 01:39:26,080 --> 01:39:28,520 GLYCAN PORTIONS, THE BUILDING 2291 01:39:28,520 --> 01:39:28,800 BLOCKS. 2292 01:39:28,800 --> 01:39:54,400 THANK YOU. 2293 01:39:54,400 --> 01:40:32,480 [ APPLAUSE ] 2294 01:40:32,480 --> 01:40:34,040 --SO THIS IS A PROJECT 2295 01:40:34,040 --> 01:40:35,440 SUPPLEMENT AS YOU HEARD FROM 2296 01:40:35,440 --> 01:40:44,480 NIKI AND I WORKING WITH 2 OTHER 2297 01:40:44,480 --> 01:40:45,240 MICROBIOLOGISTS [INDISCERNIBLE] 2298 01:40:45,240 --> 01:40:47,960 JUTRAS AND MARIA GOOD WHAT. 2299 01:40:47,960 --> 01:40:49,480 SO THE RO1 WAS BASED ON A 2300 01:40:49,480 --> 01:40:53,960 PROJECT TO BE ABLE TO DEVELOP A 2301 01:40:53,960 --> 01:40:54,920 RESERVOIR TARGETED VACCINE SO I 2302 01:40:54,920 --> 01:40:56,880 WILL STEP BACK FOR A BRIEF 2303 01:40:56,880 --> 01:40:58,640 SECOND AND SAY, THIS PROJECT IS 2304 01:40:58,640 --> 01:41:01,480 REALLY THE PERFECT POSTER CHILD 2305 01:41:01,480 --> 01:41:11,320 OF WHAT THE SUPPLEMENTS WERE 2306 01:41:11,320 --> 01:41:15,520 CREATED TO DO SO I AM A 2307 01:41:15,520 --> 01:41:17,800 VETERINARIAN WHO STUDIES DISEASE 2308 01:41:17,800 --> 01:41:20,440 IN POPULATION SAYS AND WHEN SHE 2309 01:41:20,440 --> 01:41:22,520 SAYS INVIVO SHE MEANS IN CELLS 2310 01:41:22,520 --> 01:41:24,840 AND WHEN I SAY INVIVO, I AM 2311 01:41:24,840 --> 01:41:26,720 TALKING ABOUT A CREATURE, SO I 2312 01:41:26,720 --> 01:41:28,200 AM LITERALLY THE FAR END OF THE 2313 01:41:28,200 --> 01:41:35,840 SPECTRUM LOOKING AT POPULATIONS 2314 01:41:35,840 --> 01:41:36,960 OF HUMANS, DOGS, OTHERWISE, AND 2315 01:41:36,960 --> 01:41:38,600 OF COURSE AS YOU'VE BEEN HEARING 2316 01:41:38,600 --> 01:41:40,040 FOR THE LAST DAYS YOU'RE LOOKING 2317 01:41:40,040 --> 01:41:42,240 AT GROUPS OF MOLECULES, SO IT'S 2318 01:41:42,240 --> 01:41:48,120 WONDERFUL TO BE ABLE TO BRIDGE 2319 01:41:48,120 --> 01:41:50,040 THIS TECHNOLOGY OVER TO MY 2320 01:41:50,040 --> 01:41:50,240 GROUP. 2321 01:41:50,240 --> 01:41:52,280 SO A BIT OF AN EXPLANATION OF 2322 01:41:52,280 --> 01:41:56,280 WHAT WE'RE DOING, MA RIA HAD 2323 01:41:56,280 --> 01:42:00,800 CREATED AN OUTER SURFACE PROTEIN 2324 01:42:00,800 --> 01:42:01,400 [INDISCERNIBLE] THE CAUSATIVE 2325 01:42:01,400 --> 01:42:03,320 AGENT OF LYME DISEASE, VACCINE 2326 01:42:03,320 --> 01:42:04,800 THAT WHEN MICE EAT AND WHEN MICE 2327 01:42:04,800 --> 01:42:07,240 EAT IT AS SHOWN HERE THEY THEN 2328 01:42:07,240 --> 01:42:10,040 CREATE ANTIBODIES AGAINST THAT 2329 01:42:10,040 --> 01:42:11,960 PROTEIN WHICH WHICH IS THEN 2330 01:42:11,960 --> 01:42:15,800 TAKEN UP BY THE TICK THAT 2331 01:42:15,800 --> 01:42:17,360 CARRIES LYME DISEASE. 2332 01:42:17,360 --> 01:42:19,120 THAT TICK THEN LOSES THE CANNED 2333 01:42:19,120 --> 01:42:21,040 WHAT, THEY ARE KILLED AND BY 2334 01:42:21,040 --> 01:42:23,480 CLEARING IT IDENTITY OF THE 2335 01:42:23,480 --> 01:42:26,120 TICKS, WE PROPOSE IN ON OUR RO1 2336 01:42:26,120 --> 01:42:31,040 TO SEE IF WE CAN THEN PREVENT 2337 01:42:31,040 --> 01:42:33,080 INFECTION OF BOTH HUMANS BUT AS 2338 01:42:33,080 --> 01:42:37,040 A PROXY WE ARE USING A GROUP OF 2339 01:42:37,040 --> 01:42:37,480 HUNTING DOGS. 2340 01:42:37,480 --> 01:42:40,600 IN ORDER TO DO THIS BETTER WE 2341 01:42:40,600 --> 01:42:43,440 REALLY WOULD LIKE TO HAVE BETTER 2342 01:42:43,440 --> 01:42:45,920 DIAGNOSTICS SO THE CURRENT 2343 01:42:45,920 --> 01:42:47,720 DIAGNOSTICS TO IDENTIFY 2344 01:42:47,720 --> 01:42:52,600 INFECTION WITH LYME DISEASE AND 2345 01:42:52,600 --> 01:42:54,440 SERIES POINTS OOLG BASED 2346 01:42:54,440 --> 01:42:55,120 ESPECIALLY FOR INFECTION AND 2347 01:42:55,120 --> 01:42:56,880 THESE ARE THE 1S WE WOULD LIKE 2348 01:42:56,880 --> 01:42:58,200 TO SEE THE DIFFERENCE 2349 01:42:58,200 --> 01:42:59,440 PARTICULARLY IN THE FIRST COUPLE 2350 01:42:59,440 --> 01:43:01,240 YEARS OF OUR TRIAL AND WE'RE IN 2351 01:43:01,240 --> 01:43:03,680 YEAR 3 RIGHT NOW, SO IT'S 2352 01:43:03,680 --> 01:43:06,040 PERFECT TIMING TO BE DEVELOPING 2353 01:43:06,040 --> 01:43:08,120 THESE THINGS. 2354 01:43:08,120 --> 01:43:10,840 SO JUST A QUICK REMINDER THAT 2355 01:43:10,840 --> 01:43:13,400 ALL BACTERIA HAVE THESE GLYCANS 2356 01:43:13,400 --> 01:43:14,880 AND THEY CREATE THEIR CELL WALL 2357 01:43:14,880 --> 01:43:17,960 IN ORDER TO KEEP THEM FROM 2358 01:43:17,960 --> 01:43:19,040 BURSTING THROUGH OZ METIC 2359 01:43:19,040 --> 01:43:27,200 PRESSURE, BUT WHAT'S SPECIAL 2360 01:43:27,200 --> 01:43:29,040 ABOUT BURELLIA, AS NIKI JUST 2361 01:43:29,040 --> 01:43:32,920 SAID AND THE SHORT PEPTIDES 2362 01:43:32,920 --> 01:43:33,920 WHICH DIAMEANS ARE DIFFERENT. 2363 01:43:33,920 --> 01:43:36,280 SO IF WE TALK ABOUT GRAM 2364 01:43:36,280 --> 01:43:37,600 NEGATIVE OR POSITIVE BACTERIA, 2365 01:43:37,600 --> 01:43:40,000 THE MOST COMMON TYPES THESE ARE 2366 01:43:40,000 --> 01:43:43,520 THE TYPES OF PEPTIDES YOU HAVE, 2367 01:43:43,520 --> 01:43:45,840 BUT FOR [INDISCERNIBLE], WE HAVE 2368 01:43:45,840 --> 01:43:48,720 THIS [INDISCERNIBLE] GLUE 2369 01:43:48,720 --> 01:43:49,440 MARIOUS SEEN FORMATION. 2370 01:43:49,440 --> 01:43:53,640 SO THE GOALS FOR OUR SUPPLEMENT 2371 01:43:53,640 --> 01:43:56,240 WERE TO SYNTHESIZE THIS 2372 01:43:56,240 --> 01:43:57,360 [INDISCERNIBLE] PEPTIDES AND 2373 01:43:57,360 --> 01:43:59,800 DR. PLATTENER AND HIS PNAS PAPER 2374 01:43:59,800 --> 01:44:02,440 IDENTIFIED ABOUT 13 OF THEM SO 2375 01:44:02,440 --> 01:44:06,720 WE TARGETED IT DOWN TO THE MOST 2376 01:44:06,720 --> 01:44:08,440 IMMUNOGENIC 1S BECAUSE THEY ARE 2377 01:44:08,440 --> 01:44:10,280 THE MOST USEFUL DIAGNOSTICALLY, 2378 01:44:10,280 --> 01:44:13,240 AND WE WILL CREATE THEM AS 2379 01:44:13,240 --> 01:44:15,040 REFERENCE STANDARDS FOR 2380 01:44:15,040 --> 01:44:17,560 DIAGNOSTIC PURPOSES, TO BE USED 2381 01:44:17,560 --> 01:44:20,080 AS INTERVENTIONS LIKE ELIZA, OR 2382 01:44:20,080 --> 01:44:23,880 PC R, TO LOOK AT THE ACTUAL 2383 01:44:23,880 --> 01:44:30,040 PRESENCE OR ABSENCE EVER THESE 2384 01:44:30,040 --> 01:44:33,320 PEPTIDES IN A BIOLOGICAL 2385 01:44:33,320 --> 01:44:36,440 SYMPTOMS, AS WE THINK PIE IN THE 2386 01:44:36,440 --> 01:44:38,080 SKY BIG PICTURE IF I CAN GET 2387 01:44:38,080 --> 01:44:39,440 KILOINGS OF IT AS IT WAS JUST 2388 01:44:39,440 --> 01:44:41,400 MENTIONED TO BE ABLE TO USE 2389 01:44:41,400 --> 01:44:43,240 THESE FOR IMMUNO LOGICAL ASSAYS, 2390 01:44:43,240 --> 01:44:45,920 BECAUSE WE KNOW THAT ASIDE FROM 2391 01:44:45,920 --> 01:44:48,480 ANTIBODY RESPONSES IT ALSO CAN 2392 01:44:48,480 --> 01:44:50,960 DRIVE T-CELL AND NK T-CELL 2393 01:44:50,960 --> 01:44:52,720 RESPONSES AND THESE ARE OF REAL 2394 01:44:52,720 --> 01:44:54,680 INTEREST OF MY GROUP AND 2395 01:44:54,680 --> 01:44:56,200 HONESTLY, IT'S A REAL GROWING 2396 01:44:56,200 --> 01:44:59,520 AREA IN THE FIELD OF BOTH CANCER 2397 01:44:59,520 --> 01:45:01,040 AND INFECT YOWZ IMMUNOLOGY. 2398 01:45:01,040 --> 01:45:02,960 AND THEN LASTLY, CAN YOU MAKE 2399 01:45:02,960 --> 01:45:05,360 THE SPECIFIC TARGETS FROM 2400 01:45:05,360 --> 01:45:06,120 MONOCLONAL ANTIBODY PRODUCTION 2401 01:45:06,120 --> 01:45:08,800 AND AND WE CAN USE THOSE 2402 01:45:08,800 --> 01:45:11,240 MONOCLONE'S EABT BODIES FROM 2403 01:45:11,240 --> 01:45:12,920 EVERYTHING FROM ARRAYS TO 2404 01:45:12,920 --> 01:45:14,640 SPECIFIC LATERAL FLOW ASSAYS 2405 01:45:14,640 --> 01:45:18,440 LIKE WHAT WE'RE USING TO 2406 01:45:18,440 --> 01:45:19,680 ADENTIFY SARS-COV-2. 2407 01:45:19,680 --> 01:45:21,560 SO AS YOU MIGHT IMAGINE THERE 2408 01:45:21,560 --> 01:45:23,440 WAS DELAY DUE TO PANDEMIC BUT WE 2409 01:45:23,440 --> 01:45:27,720 WERE SO PLEASED TO HAVE 2410 01:45:27,720 --> 01:45:29,240 [INDISCERNIBLE] JOIN NIKI'S 2411 01:45:29,240 --> 01:45:29,840 GROUP LAST YEAR. 2412 01:45:29,840 --> 01:45:32,720 HERE SHE IS AND MOST POST DOC, I 2413 01:45:32,720 --> 01:45:36,360 WAS ANAL TO GO TO IN THE NLS 2414 01:45:36,360 --> 01:45:38,680 JULY AND SPENT A MONTH DOING 2415 01:45:38,680 --> 01:45:39,760 BASIC TRAINING WITH THE GROUP TO 2416 01:45:39,760 --> 01:45:44,320 LEARN SO SHE WAS A MOLECULAR BI 2417 01:45:44,320 --> 01:45:45,000 BIOLOGIST BY TRANSLATIONAL 2418 01:45:45,000 --> 01:45:46,440 RESEARCH TRAINING AND TO LEARN 2419 01:45:46,440 --> 01:45:48,960 MORE ABOUT THE MANUEL SYNTHESIS 2420 01:45:48,960 --> 01:45:52,240 FOR THESE NEUROPEPTIDES I ALSO 2421 01:45:52,240 --> 01:45:53,480 ENJOYED DR. WILSON'S TALK 2422 01:45:53,480 --> 01:45:54,760 YESTERDAY AND OF COURSE NOW 2423 01:45:54,760 --> 01:45:57,000 BECAUSE THE GOAL IS TO HAVE AN 2424 01:45:57,000 --> 01:46:00,240 AUTOMATED SYNTHESIS FOR THESE 2425 01:46:00,240 --> 01:46:01,320 DIFFERENT PEPTIDES, 2426 01:46:01,320 --> 01:46:01,640 NEUROPEPTIDES. 2427 01:46:01,640 --> 01:46:03,760 BUT FOR NOW WE NEED TO FIGURE 2428 01:46:03,760 --> 01:46:06,080 OUT HOW TO MAKE THEM, THE FIRST 2429 01:46:06,080 --> 01:46:08,840 TIME, SO THIS IS WHAT THE GROUP 2430 01:46:08,840 --> 01:46:12,560 HAS BEEN WORKING ON AS WELL AS 2431 01:46:12,560 --> 01:46:13,280 [INDISCERNIBLE]. 2432 01:46:13,280 --> 01:46:13,840 HERE. 2433 01:46:13,840 --> 01:46:15,480 AND AFTER THAT TIME, YOU KNOW 2434 01:46:15,480 --> 01:46:18,040 THE 3 OF THEM WOG TOGETHER TO 2435 01:46:18,040 --> 01:46:19,480 TRULY IDENTIFY WHAT ARE 2436 01:46:19,480 --> 01:46:20,840 NEUROPEPTIDES OF INTEREST AND 2437 01:46:20,840 --> 01:46:26,080 THE BIG 1 IS SHOWN HERE. 2438 01:46:26,080 --> 01:46:29,160 SO, THIS WAS IDENTIFIED BY 2439 01:46:29,160 --> 01:46:30,600 BRANDON AND DR. [INDISCERNIBLE] 2440 01:46:30,600 --> 01:46:33,760 LAB TO BE HIGH OWE IMMUNOGENIC 2441 01:46:33,760 --> 01:46:35,960 FOUND IN PATIENTS WITH LYME 2442 01:46:35,960 --> 01:46:40,960 ARTHRITIS AND BECAUSE OF THAT 2443 01:46:40,960 --> 01:46:42,720 THIS HYDROMIEWRIN AC IS OUR 2444 01:46:42,720 --> 01:46:45,600 HIGHEST TARGET IN ORD TO DETECT 2445 01:46:45,600 --> 01:46:46,440 IT, THEY'RE STILL FINISHING THE 2446 01:46:46,440 --> 01:46:48,560 SCREAMS FOR THIS IS SYNTHESIZING 2447 01:46:48,560 --> 01:46:51,640 IT SO WE STARTED IDENTIFYING THE 2448 01:46:51,640 --> 01:46:54,040 TRI PEPTIDE AS A FIRST STEP IN 2449 01:46:54,040 --> 01:46:54,600 MY LAB. 2450 01:46:54,600 --> 01:46:57,000 SO THIS WAS SENT TO US IN 2451 01:46:57,000 --> 01:46:57,240 FEBRUARY. 2452 01:46:57,240 --> 01:47:01,120 AND HOT OFF THE PRESSES, THIS IS 2453 01:47:01,120 --> 01:47:02,640 OUR MS TEAM AT THE UNIVERSITY OF 2454 01:47:02,640 --> 01:47:05,280 IOWA AND TO REMIND YOU WE DON'T 2455 01:47:05,280 --> 01:47:06,720 RUN MS ALL THE TIME. 2456 01:47:06,720 --> 01:47:09,640 WE ARE RUNNING FACTS ASHESINAL 2457 01:47:09,640 --> 01:47:13,800 SIS AND ELIZAS AND PERFORMING 2458 01:47:13,800 --> 01:47:15,240 HIGHER LEVEL EPIDEMIOLOGICAL 2459 01:47:15,240 --> 01:47:16,400 STATISTICAL ANALYSIS IN OUR 2460 01:47:16,400 --> 01:47:18,040 POPULATION, SO MS WAS NOT 2461 01:47:18,040 --> 01:47:19,440 SOMETHING WE NORMALLY DO AND 2462 01:47:19,440 --> 01:47:21,520 IT'S FANTASTIC TO SHOW YOU THIS 2463 01:47:21,520 --> 01:47:22,600 DATA SIMPLE THOUGH IT MIGHT BE 2464 01:47:22,600 --> 01:47:25,600 SO WE WERE LOOKING AT THE 2465 01:47:25,600 --> 01:47:27,000 MASTERY CHARGE RATIOS WHICH WERE 2466 01:47:27,000 --> 01:47:27,760 CONSIST WENT WHAT WE'RE 2467 01:47:27,760 --> 01:47:30,400 EXPECTING BRACED ON THE DATA OUT 2468 01:47:30,400 --> 01:47:34,160 OF THE POHL LAB AND WE WERE ABLE 2469 01:47:34,160 --> 01:48:33,440 TO THEN SEE THAT THAT EVEN 2470 01:48:33,440 --> 01:48:37,600 THOUGH WE AND LIKE--LIKE I SAID 2471 01:48:37,600 --> 01:48:39,680 WITH LINKAGES, WHEN WE LOOK AT 2472 01:48:39,680 --> 01:48:42,760 THESE PEAKS, WE CAN FIND THE TRI 2473 01:48:42,760 --> 01:48:45,720 PECTIDE MSMS IN HERE, AND 2474 01:48:45,720 --> 01:48:46,680 INDICATING THIS SPECIFICS TRI 2475 01:48:46,680 --> 01:48:49,640 PEPTIDE THAT IS FOUND IN 2476 01:48:49,640 --> 01:48:51,040 [INDISCERNIBLE], SO THE NEXT 2477 01:48:51,040 --> 01:48:55,360 STEPS ARE TO COMPLETE THE POETIC 2478 01:48:55,360 --> 01:48:56,040 TEBTIAL [INDISCERNIBLE] TARGETS, 2479 01:48:56,040 --> 01:49:01,080 COMBINE THEM ALL AND LOOK AT AND 2480 01:49:01,080 --> 01:49:01,760 FRAGMENTATION PATTERNS, THEN 2481 01:49:01,760 --> 01:49:03,160 WE'RE GOING TO START SOME ASSAYS 2482 01:49:03,160 --> 01:49:06,880 TO REALLY SEE HOW WELL THIS 2483 01:49:06,880 --> 01:49:08,960 WORKS AS I DIAGNOSTIC, WE WANT 2484 01:49:08,960 --> 01:49:11,440 TO COMPARE IT TO SAMPLES FROM 2485 01:49:11,440 --> 01:49:13,280 OTHER COMMON PACTERRIA THAT ARE 2486 01:49:13,280 --> 01:49:15,360 LIKELY TO CONTAMINATE, LIKE 2487 01:49:15,360 --> 01:49:16,640 E.COLI OR CANNED WHAT TO SHOW 2488 01:49:16,640 --> 01:49:18,840 THE FES FISCHERITY OF THIS, AND 2489 01:49:18,840 --> 01:49:20,840 THEN, WE'RE GOING TO GET INTO 2490 01:49:20,840 --> 01:49:27,360 THE ACTUAL SERUM SAMPLES FROM 2491 01:49:27,360 --> 01:49:27,600 OUR TRIAL. 2492 01:49:27,600 --> 01:49:29,600 AND BOTH FIRST WITH HUMAN SERUM 2493 01:49:29,600 --> 01:49:30,960 AND DIFFERENT COMPOUNDS AND THEN 2494 01:49:30,960 --> 01:49:32,440 GET INTO ANALYZING OUR SPECIFIC 2495 01:49:32,440 --> 01:49:34,120 SAMPLE SO WE'RE EXCITED ABOUT 2496 01:49:34,120 --> 01:49:35,840 THIS AND IT'S BEEN A REAL 2497 01:49:35,840 --> 01:49:37,840 PLEASURE TO WORK WITH DR. POHL. 2498 01:49:37,840 --> 01:49:57,920 THANK YOU ALL FOR YOUR TIME AND 2499 01:49:57,920 --> 01:49:58,200 ATTENTION. 2500 01:49:58,200 --> 01:49:59,360 NTHANK YOU VERY MUCH AND OUR 2501 01:49:59,360 --> 01:50:01,040 FINAL TOWK THIS MORNING BEFORE A 2502 01:50:01,040 --> 01:50:19,280 BRIEF BREAK IS DR. DAVID CRICH. 2503 01:50:19,280 --> 01:50:21,040 >> MORNING EVERYONE AND THANK 2504 01:50:21,040 --> 01:50:29,080 FOR LISTENING TO MY TALK AND 2505 01:50:29,080 --> 01:50:31,320 ALSO THANKS FOR THE NIH FOR 2506 01:50:31,320 --> 01:50:33,760 PUTTING TOGETHER THIS TREMENDOUS 2507 01:50:33,760 --> 01:50:34,240 PROGRAM. 2508 01:50:34,240 --> 01:50:35,120 I'M HERE REPRESENTING AND 2509 01:50:35,120 --> 01:50:36,880 MYSELF, WE DON'T HAVE ANY 2510 01:50:36,880 --> 01:50:40,080 CONFLICTS OF INTEREST THAT WE 2511 01:50:40,080 --> 01:50:53,640 KNOW OF. 2512 01:50:53,640 --> 01:50:54,080 SO, OKAY. 2513 01:50:54,080 --> 01:51:00,600 LET ME GO BACK 1. 2514 01:51:00,600 --> 01:51:02,480 OKAY, SO SO LIKE EVERYONE ELSE 2515 01:51:02,480 --> 01:51:04,440 WE INSPIRED BY THE PUBLICATION 2516 01:51:04,440 --> 01:51:05,840 OF THE NATIONAL ACADEMIES OF 2517 01:51:05,840 --> 01:51:07,760 SCIENCE AND WE USE THIS TO 2518 01:51:07,760 --> 01:51:11,440 STRUCTURE OUR PROGRAM, SO, WE 2519 01:51:11,440 --> 01:51:16,160 PAID PARTICULAR ATTENTION TO 2520 01:51:16,160 --> 01:51:19,040 THESE 2 COMMENTS HERE ABOUT THE 2521 01:51:19,040 --> 01:51:19,720 REPRODUCIBILITY OF GLYCOSYLATION 2522 01:51:19,720 --> 01:51:23,640 REACTIONS AND THE NEED TO DO 2523 01:51:23,640 --> 01:51:24,160 SOMETHING ABOUT THAT. 2524 01:51:24,160 --> 01:51:27,360 AND THEN WE ALSO PAID ATTENTION 2525 01:51:27,360 --> 01:51:31,680 TO THE NEED TO BRING IT INTO THE 2526 01:51:31,680 --> 01:51:33,240 MAIN STREAM SYNTHETIC 2527 01:51:33,240 --> 01:51:34,600 ORGANIZATIONS GANNAIC CHEMISTRY, 2528 01:51:34,600 --> 01:51:36,280 BE 1 WAY TO GO BA THIS OF COURSE 2529 01:51:36,280 --> 01:51:39,040 IS TO LOOK AT NEW GLYCOSYLATION 2530 01:51:39,040 --> 01:51:40,240 METHODS AND THAT CERTAINLY HAS 2531 01:51:40,240 --> 01:51:44,640 BEEN A FOCUS OF OUR UR1 PROGRAM 2532 01:51:44,640 --> 01:51:48,440 AND IT'S DIVIDED INTO VARIOUS 2533 01:51:48,440 --> 01:51:51,840 DIFFERENT SPECIFIC AIMS, SO MY 2534 01:51:51,840 --> 01:51:53,160 COLLABORATOR PETER GAVE A POSTER 2535 01:51:53,160 --> 01:51:55,400 YESTERDAY ON WHAT WE'VE BEEN 2536 01:51:55,400 --> 01:51:57,160 DOING WITH MICROWAVE ASSISTED 2537 01:51:57,160 --> 01:51:59,360 DPLI COSALATION, I'M VERY 2538 01:51:59,360 --> 01:52:01,640 INTERESTED IN PHOTO CHEMICAL 2539 01:52:01,640 --> 01:52:05,000 GLYCOSYLATION AND I--I DO THINK 2540 01:52:05,000 --> 01:52:07,600 THIS IS A WAY FORWARD AND I DO 2541 01:52:07,600 --> 01:52:09,480 HAVE NICE RESULTS THERE, BUT I'M 2542 01:52:09,480 --> 01:52:11,240 NOT READY TO TALK ABOUT THEM 2543 01:52:11,240 --> 01:52:11,440 YET. 2544 01:52:11,440 --> 01:52:13,360 WE WILL JUST CALL THAT A WORK IN 2545 01:52:13,360 --> 01:52:16,440 PROGRESS AND LEAVE IT THERE, AND 2546 01:52:16,440 --> 01:52:18,320 THEN, ANOTHER THEME OF OUR 2547 01:52:18,320 --> 01:52:20,480 PROJECT HAS BEEN THE DEVELOPMENT 2548 01:52:20,480 --> 01:52:22,400 OF METHODS THROUGH THE BACTERIAL 2549 01:52:22,400 --> 01:52:24,360 GLYCANS AND I HAVE BEEN WORKING 2550 01:52:24,360 --> 01:52:26,320 ON THIS FOR MANY YEARS AS WELL. 2551 01:52:26,320 --> 01:52:32,040 SO I WILL JUST GIVE YOU I 2552 01:52:32,040 --> 01:52:33,600 SNAPSHOT OF WHAT WE'VE BEEN 2553 01:52:33,600 --> 01:52:34,880 DOING REEBTLY SO YOU DON'T THINK 2554 01:52:34,880 --> 01:52:36,920 THAT I'VE GONE INTO RETIREMENT, 2555 01:52:36,920 --> 01:52:52,440 SO, A FEW YEARS AGO, WE GOT INTO 2556 01:52:52,440 --> 01:52:55,520 THE LEGION AMINIC ACID AND 2557 01:52:55,520 --> 01:52:56,840 PSEUDOMINNIC ACID, SO WE 2558 01:52:56,840 --> 01:52:57,880 DESIGNED THIS DONEAR AND 2559 01:52:57,880 --> 01:53:01,520 ACTUALLY WE SHOWED THAT IT 2560 01:53:01,520 --> 01:53:04,400 WORKED REALLY NICELY AND WITH 2561 01:53:04,400 --> 01:53:05,880 THE PSEUDOMENNIC ACID FOR THIS 2562 01:53:05,880 --> 01:53:08,640 DONOR HERE AND WE SHOWED THAT IT 2563 01:53:08,640 --> 01:53:10,840 WORKED EVEN BETTER SO BOTH OF 2564 01:53:10,840 --> 01:53:13,040 THESE CLASSES OF GLYCO SITES CAN 2565 01:53:13,040 --> 01:53:16,800 BE MADE NOW, THE PROBLEM IS 2566 01:53:16,800 --> 01:53:25,640 THATY YOU HAVE TO MAKE THE 2567 01:53:25,640 --> 01:53:28,640 DONORS AND THE DONOR WE USE 20 2568 01:53:28,640 --> 01:53:30,960 STEPS, SO WE DON'T THINK THAT 2569 01:53:30,960 --> 01:53:33,200 ANYONE IS ABOUT TO DO THAT 2570 01:53:33,200 --> 01:53:33,840 ANYTIME SOON. 2571 01:53:33,840 --> 01:53:36,040 SO CLEARLY WE HAVE BEEN WORKING 2572 01:53:36,040 --> 01:53:39,880 TO IMPROVE THIS, AND SO, THIS IS 2573 01:53:39,880 --> 01:53:42,280 WHAT WE PUBLISHED JUST A COUPLE 2574 01:53:42,280 --> 01:53:46,000 WEEKS AGO, SO WE STILL GO OUT 2575 01:53:46,000 --> 01:53:47,640 FROM [INDISCERNIBLE] ACID, AND 2576 01:53:47,640 --> 01:53:49,480 WE HAVE DESIGNED MUCH BETTER 2577 01:53:49,480 --> 01:53:53,040 CHEMISTRY NOW TO MAKE THE REGION 2578 01:53:53,040 --> 01:53:57,560 DONOR AND THE TRICK THAT WE USE 2579 01:53:57,560 --> 01:53:59,520 HERE BURN OFF THE SIDE CHAIN 2580 01:53:59,520 --> 01:54:01,760 FROM THIS TRIAL TO MAKE AN ALD 2581 01:54:01,760 --> 01:54:05,800 EHIDE HERE AND THEN WE USE THE 2582 01:54:05,800 --> 01:54:08,280 CHEMISTRY SO WE HAVE THIS WITH 2583 01:54:08,280 --> 01:54:11,080 12 STEPS, IT'S ACTUALLY FAIRLY 2584 01:54:11,080 --> 01:54:14,640 PRACTICAL AND IN ALL HONESTY, 2585 01:54:14,640 --> 01:54:16,600 THIS MATERIAL HERE CAN BE MADE 2586 01:54:16,600 --> 01:54:21,360 ON SCALE AND COULD AND SHOULD BE 2587 01:54:21,360 --> 01:54:21,640 SPECIALIZED. 2588 01:54:21,640 --> 01:54:23,000 AN YOU HAVE TO SHOOT OF THIS IS 2589 01:54:23,000 --> 01:54:26,440 ALSO THAT WE'RE ABLE TO MAKE 2590 01:54:26,440 --> 01:54:28,080 ACETA MINNIC ACID ALSO IN TWEP 2591 01:54:28,080 --> 01:54:29,240 STEPS AND MAKE THAT AVAILABLE, 2592 01:54:29,240 --> 01:54:32,880 SO WE HAVE ABOUT HALF A GRAM OF 2593 01:54:32,880 --> 01:54:35,160 THIS AROUND NOW AND THEN 2594 01:54:35,160 --> 01:54:36,440 PSEUDOMENNIC ACID, THE MOMENT WE 2595 01:54:36,440 --> 01:54:40,040 HAVE IT DOWN TO 15 STEPS, USING 2596 01:54:40,040 --> 01:54:43,120 A VARIATION ON THE SAME KIND OF 2597 01:54:43,120 --> 01:54:45,240 CHEMISTRY, BURN OFF THE SIDE 2598 01:54:45,240 --> 01:54:46,760 CHAIN AND THEN REINSTALL THE 2599 01:54:46,760 --> 01:54:47,240 PIECES. 2600 01:54:47,240 --> 01:54:48,920 THE EXTRA STEPS COME FROM THE 2601 01:54:48,920 --> 01:54:52,000 NEED TO PUT IN THIS AXIAL CN 2602 01:54:52,000 --> 01:54:55,080 BOND THERE, SO THIS CHEMISTRY IS 2603 01:54:55,080 --> 01:54:56,560 BECOMING PRACTICAL, WE AGAIN 2604 01:54:56,560 --> 01:54:58,000 HAVE SEVERAL HUNDREDS OF 2605 01:54:58,000 --> 01:55:02,040 MILLIGRAMS OF THIS COMPOUND. 2606 01:55:02,040 --> 01:55:06,160 I WANT TO COME BACK TO THIS AND 2607 01:55:06,160 --> 01:55:07,560 THESE STATEMENTS HERE AND I 2608 01:55:07,560 --> 01:55:11,280 REALLY WANT TOED TO TELL YOU 2609 01:55:11,280 --> 01:55:12,680 ABOUT MY LONG STANDING 2610 01:55:12,680 --> 01:55:17,360 PERSPECTIVE ON THIS FIELD AS A 2611 01:55:17,360 --> 01:55:18,640 MAIN STREAM ORGANIC KRIMMIST I 2612 01:55:18,640 --> 01:55:20,960 MADE SAY AND I JUST SAID MYSELF 2613 01:55:20,960 --> 01:55:22,800 AGAIN DESCRIBED AS PHYSICAL 2614 01:55:22,800 --> 01:55:26,600 ORGANIC CHEMIST IS ALWAYS A 2615 01:55:26,600 --> 01:55:26,920 COMPLEMENT. 2616 01:55:26,920 --> 01:55:30,640 EVEN AMUSING BUT I'M JUST AN 2617 01:55:30,640 --> 01:55:32,600 ORGANIC CHEMIST, AND MY 2618 01:55:32,600 --> 01:55:33,440 PERECTOMYOSINNIVE HAS ALWAYS 2619 01:55:33,440 --> 01:55:35,200 BEEN THAT RATIONAL ADVANCES 2620 01:55:35,200 --> 01:55:38,640 DERIVED FROM A BETTER 2621 01:55:38,640 --> 01:55:40,200 APPROXIMATE INSIGHT INTO THE 2622 01:55:40,200 --> 01:55:41,320 MECHANISM OF GLYCOSYLATION AND 2623 01:55:41,320 --> 01:55:46,680 I'VE BEEN WRITING THIS IN MY NIH 2624 01:55:46,680 --> 01:55:47,640 PROPOSALS CONSISTENTLY FOR 25 2625 01:55:47,640 --> 01:55:51,840 YEARS NOW, SO WE'VE BEEN AT THIS 2626 01:55:51,840 --> 01:55:56,160 FOR A WHILE AND WHERE ALL OF 2627 01:55:56,160 --> 01:55:58,680 THIS HAS TAKEN ME IS TO THE 2628 01:55:58,680 --> 01:56:00,240 STATEMENT THAT SM1 MECHANISMS 2629 01:56:00,240 --> 01:56:03,040 STATEMENTS NEED TO BE 2630 01:56:03,040 --> 01:56:03,560 DEEMPHASIZED, STRONGLY 2631 01:56:03,560 --> 01:56:06,120 DEEMPHASIZED YOU MIGHT SAY AND I 2632 01:56:06,120 --> 01:56:11,200 HAVE TALKED ABOUT THIS TO 2633 01:56:11,200 --> 01:56:11,680 VARIOUS REVIEW ARTICLES. 2634 01:56:11,680 --> 01:56:13,560 SO WE DON'T HAVE TIME TO GO INTO 2635 01:56:13,560 --> 01:56:18,040 ALL OF THE EVIDENCE BUT HERE'S A 2636 01:56:18,040 --> 01:56:20,040 GENERAL GLYCOSYLATION METHOD AND 2637 01:56:20,040 --> 01:56:23,120 FOR THE LONGEST TIME THE FIELD 2638 01:56:23,120 --> 01:56:26,640 HAVE BEEN OPERATING HERE ON THE 2639 01:56:26,640 --> 01:56:28,520 ASSUMPTION THAT GLYCOSYLATION 2640 01:56:28,520 --> 01:56:32,200 REACTIONS PROCEED VIA SM1 2641 01:56:32,200 --> 01:56:33,720 MECHANISMS, ON NAKED OXOCARBIN O 2642 01:56:33,720 --> 01:56:36,160 MIDS AND THIS IS WHERE WE GO 2643 01:56:36,160 --> 01:56:36,400 WRONG. 2644 01:56:36,400 --> 01:56:41,240 THIS IS THE CORE OF IT AND WHY 2645 01:56:41,240 --> 01:56:44,440 GLYCOSYLATIONS REACTIONS ARE NOT 2646 01:56:44,440 --> 01:56:45,000 REPRODUCIBLE. 2647 01:56:45,000 --> 01:56:46,440 THE REALITY IS THAT 2648 01:56:46,440 --> 01:57:07,400 GLYCOSYLATIONS REACT ON A SCALE 2649 01:57:07,400 --> 01:57:09,080 AND TO KEEP THEM THERE. TO DO 2650 01:57:09,080 --> 01:57:17,400 THIS WE HAVE TO HAVE THE COUNTER 2651 01:57:17,400 --> 01:57:18,880 IONS ARE TRIPLETS SO WOE DON'T 2652 01:57:18,880 --> 01:57:20,400 HAVE TIME FOR ALL THE EVIDENCE 2653 01:57:20,400 --> 01:57:21,800 SO WITH THE COMMON FUND PROGRAM 2654 01:57:21,800 --> 01:57:24,440 WE WROTE WITH MY COWORKERS 2 2655 01:57:24,440 --> 01:57:27,720 LARGE CHEMICAL REVIEWS THAT 2656 01:57:27,720 --> 01:57:28,480 BRING TOGETHER ALL OF THE 2657 01:57:28,480 --> 01:57:30,560 EVIDENCE AND SUMMARIZE IT, SO I 2658 01:57:30,560 --> 01:57:31,720 RECOMMEND THAT YOU READ THEM AND 2659 01:57:31,720 --> 01:57:35,160 THINK ABOUT WHAT THEY SAY. 2660 01:57:35,160 --> 01:57:37,840 AND THEN, IT'S NICE TO NOTE, 2661 01:57:37,840 --> 01:57:41,240 THAT SOME OF THE MAIN ADVANCES 2662 01:57:41,240 --> 01:57:42,360 IN SYNTHETIC CHEMISTRY, COMING 2663 01:57:42,360 --> 01:57:44,800 OUT OF THIS COMMON FUND PROGRAM 2664 01:57:44,800 --> 01:57:48,600 BY COMMON FUND MEMBERS, ARE ALL 2665 01:57:48,600 --> 01:57:50,800 HIGHLY STEREO SELECTIVE DPLI 2666 01:57:50,800 --> 01:57:52,200 COSALATION REACTIONS AND AT THE 2667 01:57:52,200 --> 01:57:57,560 END OF THE DAY, THEY ALL INVOLVE 2668 01:57:57,560 --> 01:58:07,440 SM2 LIKE MECHANISMS. 2669 01:58:07,440 --> 01:58:09,160 SO THAT AND PUSH THESE REACTIONS 2670 01:58:09,160 --> 01:58:10,400 TO THE EXTREMES WE DEFINITELY 2671 01:58:10,400 --> 01:58:13,640 NEED TO STOP THINKING ABOUT 2672 01:58:13,640 --> 01:58:15,080 THINGS LIKE THIS, AND WE HAVE TO 2673 01:58:15,080 --> 01:58:17,320 NOTED REALLY IS HOW TO SHIFT 2674 01:58:17,320 --> 01:58:18,640 THESE EQUILL LIBRARY FOUNDATION 2675 01:58:18,640 --> 01:58:21,080 RIIA FROM A BETA DONOR THAT 2676 01:58:21,080 --> 01:58:22,680 GIVES AN ALPHA GEICO SEED TO AN 2677 01:58:22,680 --> 01:58:24,800 ALPHA DONOR THAT GIVES A BETA 2678 01:58:24,800 --> 01:58:27,640 FLIEKOCIDE AND IF WE CAN 2679 01:58:27,640 --> 01:58:29,640 UNDERSTAND THAT, PROPERLY, THEN 2680 01:58:29,640 --> 01:58:32,400 WE CAN DESIGN HIGHLY STEREO 2681 01:58:32,400 --> 01:58:34,960 SELECTIVE REACTIONS, SO FOR OUR 2682 01:58:34,960 --> 01:58:40,600 COWORKERS WE HAVE VARIOUS 2683 01:58:40,600 --> 01:58:43,040 LESSONS. 2684 01:58:43,040 --> 01:58:45,960 PLEASE STOP [INDISCERNIBLE]. 2685 01:58:45,960 --> 01:58:46,640 EVEN MECHANISTIC ENZYMOLOGISTS 2686 01:58:46,640 --> 01:58:48,760 DON'T DO THAT BUT YET WE 2687 01:58:48,760 --> 01:58:49,080 PERSISTED IT. 2688 01:58:49,080 --> 01:58:52,200 WE NEED TO THINK IN TERMS OF 2689 01:58:52,200 --> 01:58:54,960 CONCENTRATION AND NOTED 2690 01:58:54,960 --> 01:58:56,680 EQUIVALENT BECAUSE WE NEED TO 2691 01:58:56,680 --> 01:58:58,160 UNDERSTAND THE DIFFERENCE 2692 01:58:58,160 --> 01:59:01,240 BETWEEN THE RATE LAW FOR AN AN1 2693 01:59:01,240 --> 01:59:04,680 REACTION AND AN SN2 REACTION AND 2694 01:59:04,680 --> 01:59:08,520 THE DIFFERENCES IN THE WAY THEY 2695 01:59:08,520 --> 01:59:09,320 DEPEND ON CONCENTRATION AND IF 2696 01:59:09,320 --> 01:59:11,160 YOU THINK ABOUT THAT, IT TELLS 2697 01:59:11,160 --> 01:59:12,400 YOU THAT AS RAY ACTION 2698 01:59:12,400 --> 01:59:14,640 PROGRESSES AND THE 2699 01:59:14,640 --> 01:59:16,560 CONCENTRATIONS DIMINISH, THEN 2700 01:59:16,560 --> 01:59:19,520 THE MECHANISM SHIFTS OVER HERE 2701 01:59:19,520 --> 01:59:21,040 AND BECOMES MORE SN1 LIKE, 2702 01:59:21,040 --> 01:59:24,440 DURING THE COURSE OF THE 2703 01:59:24,440 --> 01:59:25,640 REACTION. 2704 01:59:25,640 --> 01:59:27,760 AND THAT'S--THAT'S AN IMPORTANT 2705 01:59:27,760 --> 01:59:28,040 REALIZATION. 2706 01:59:28,040 --> 01:59:32,520 AND THE ORDER TO COMBAT THAT, WE 2707 01:59:32,520 --> 01:59:34,120 SHOULD WORK ON THE FIRST ORDER 2708 01:59:34,120 --> 01:59:36,240 CONDITIONS AND THEN WE SORT OF 2709 01:59:36,240 --> 01:59:38,000 NEED TO REALLY THINK ABOUT THE 2710 01:59:38,000 --> 01:59:40,320 COUNTER IONS AND HOW TO PISH 2711 01:59:40,320 --> 01:59:41,560 THESE EQUILL LIBRARY FOUNDATION 2712 01:59:41,560 --> 01:59:46,880 Y RIA TO THIS SEAD OR THIS SIDE. 2713 01:59:46,880 --> 01:59:48,520 SO TRYING TO FORMALIZE ALL THIS 2714 01:59:48,520 --> 01:59:51,080 AND GET THIS MESSAGE ACROSS TO A 2715 01:59:51,080 --> 01:59:53,360 BROAD AUDIENCE, PETER AND I WE 2716 01:59:53,360 --> 01:59:54,960 PUT TOGETHER THESE FIED LINES 2717 01:59:54,960 --> 01:59:57,760 FOR OLD GRIEK O SIDE FORMATION 2718 01:59:57,760 --> 02:00:00,960 NAFIRST PRINCIPLES THAT WE 2719 02:00:00,960 --> 02:00:03,200 PUSHLISHED LAST YEAR IN ACS 2720 02:00:03,200 --> 02:00:03,560 CENTRAL SCIENCE. 2721 02:00:03,560 --> 02:00:06,040 SO CAN YOU SEE THAT WE PUT 2722 02:00:06,040 --> 02:00:10,120 TOGETHER 12 GUIDELINES FOR DOING 2723 02:00:10,120 --> 02:00:11,640 SEROSELECTIVE GLYCOSYLATION AND 2724 02:00:11,640 --> 02:00:14,000 THE MAIN FEATURES ARE 2725 02:00:14,000 --> 02:00:14,640 CONCENTRATION, TEMPERATURE, AND 2726 02:00:14,640 --> 02:00:17,280 THE COUNTER ION AND THESE 2727 02:00:17,280 --> 02:00:19,680 GUIDELINES WERE DEVELOPED TO 2728 02:00:19,680 --> 02:00:26,880 DIFFERENT PLACES ON I75 OR IN 2729 02:00:26,880 --> 02:00:30,400 PROXIMITY TO IT, AND TO KEEP THE 2730 02:00:30,400 --> 02:00:31,480 TEMPERATURE DOWN, OF THE YIELD 2731 02:00:31,480 --> 02:00:34,000 OF THE SELECTIVITY AND TAKE NOTE 2732 02:00:34,000 --> 02:00:34,640 OF THE COUNTERION. 2733 02:00:34,640 --> 02:00:40,200 THOSE ARE THE CORE MESSAGES, SO, 2734 02:00:40,200 --> 02:00:43,840 GUIDELINE 1 IS FAIRLY OBVIOUS, 2735 02:00:43,840 --> 02:00:45,880 WE NEED TO KEEP GOOD 2736 02:00:45,880 --> 02:00:47,520 EXPERIMENTAL BOTS, AND IN 2737 02:00:47,520 --> 02:00:50,240 PARTICULAR, WE NEED TO DISCUSS 2738 02:00:50,240 --> 02:00:52,680 THE CONCENTRATION AND THE AMOUNT 2739 02:00:52,680 --> 02:00:54,760 OF SOLVENT THAT WE USE. 2740 02:00:54,760 --> 02:00:58,160 THEN, IN ORDER TO DO 2741 02:00:58,160 --> 02:00:58,920 COMPARISONS, WE SHOULD--WE 2742 02:00:58,920 --> 02:01:00,680 SHOULD NOTE THAT WE SHOULD--WE 2743 02:01:00,680 --> 02:01:03,240 SHOULD PICK A STANDARD 2744 02:01:03,240 --> 02:01:06,080 CONVERSION OR PRODUCT YIELD, WE 2745 02:01:06,080 --> 02:01:08,400 SUGGEST AN 80% YIELD AND IF 2746 02:01:08,400 --> 02:01:11,000 EVERYONE REPORTED REACTIONS AT 2747 02:01:11,000 --> 02:01:13,600 THE SAME CONVERSION, IT WOULD 2748 02:01:13,600 --> 02:01:14,840 MAKE COMPARISON BETWEEN 2749 02:01:14,840 --> 02:01:17,720 DIFFERENT METHODS MUCH EASIER 2750 02:01:17,720 --> 02:01:21,440 AND MUCH MORE ACCURATE. 2751 02:01:21,440 --> 02:01:22,600 BECAUSE OF THE CONCENTRATION 2752 02:01:22,600 --> 02:01:25,240 PROBLEM WE SHOULD WORK UNDER THE 2753 02:01:25,240 --> 02:01:26,040 PSEUDOFIRST ORDER CONDITIONS AND 2754 02:01:26,040 --> 02:01:27,240 BIG AT THAT TIME DAILY BASIS 2755 02:01:27,240 --> 02:01:30,200 APPROACHES TO MAKING PREDICTIONS 2756 02:01:30,200 --> 02:01:32,280 SHOULD ONLY DUCE THAT DATA THAT 2757 02:01:32,280 --> 02:01:37,120 HAS THE CONCENTRATION IN IT. 2758 02:01:37,120 --> 02:01:38,520 A SECOND IMPORTANT THING IS 2759 02:01:38,520 --> 02:01:38,840 TEMPERATURE. 2760 02:01:38,840 --> 02:01:40,760 AGAIN IF YOU THINK OF THE 2761 02:01:40,760 --> 02:01:42,120 DIFFERENT ENTROPY REQUIREMENTS 2762 02:01:42,120 --> 02:01:44,040 FOR THE TRANSITION STATE OF AN 2763 02:01:44,040 --> 02:01:47,520 SN 1, AND AN SN2 REACTION, IT'S 2764 02:01:47,520 --> 02:01:50,160 OBVIOUS THAT IF YOU ALLOW THE 2765 02:01:50,160 --> 02:01:51,640 TEMPERATURE TO CHANGE DURING THE 2766 02:01:51,640 --> 02:01:54,240 COURSE OF A REACTION, THE 2767 02:01:54,240 --> 02:01:54,840 MECHANISM WILL CHANGE. 2768 02:01:54,840 --> 02:01:58,560 SO WHAT WE NEED TO DO IS TO 2769 02:01:58,560 --> 02:02:01,200 OPERATE OUR REACTIONS AT A 2770 02:02:01,200 --> 02:02:01,920 CONSTANT TEMPERATURE AND THAT IF 2771 02:02:01,920 --> 02:02:05,280 WE WANT TO DO THE SN2 LIKE 2772 02:02:05,280 --> 02:02:06,920 MECHANISMS, THEN, THE REACTION 2773 02:02:06,920 --> 02:02:10,640 SHOULD BE CONDUCTED AT THE 2774 02:02:10,640 --> 02:02:11,320 LOWEST POSSIBLE TEMPERATURE 1 2775 02:02:11,320 --> 02:02:14,920 CYST WENT OUR PRACTICAL'S ACTION 2776 02:02:14,920 --> 02:02:18,600 TIME AND THEN WE HAVE TO THINK 2777 02:02:18,600 --> 02:02:21,360 ABOUT COUNTER IONS AND SO ON AND 2778 02:02:21,360 --> 02:02:21,960 SO FORTH. 2779 02:02:21,960 --> 02:02:25,360 AND IT SO HAPPENS THAT 2780 02:02:25,360 --> 02:02:26,840 [INDISCERNIBLE] IS A GOOD 2781 02:02:26,840 --> 02:02:39,840 COUNTER ION, --IF FACT WE HAD TO 2782 02:02:39,840 --> 02:02:42,760 TALK ABOUT THE ACCEPTAR AND 2783 02:02:42,760 --> 02:02:44,280 MAKING IT MORE NUCLEOPHILIC AND 2784 02:02:44,280 --> 02:02:46,440 PROTECTING GROUPS AND SO ON, 2785 02:02:46,440 --> 02:02:48,640 THEY LEAD TO COMPLICATIONS AND 2786 02:02:48,640 --> 02:02:52,880 THEN, WE SHOULD WORRY ABOUT 2787 02:02:52,880 --> 02:02:54,400 SOLVENT AND DISCIPLINARY CLOOR 2788 02:02:54,400 --> 02:02:56,840 MEAN IS ULTIMATE, SO WE 2789 02:02:56,840 --> 02:02:57,680 PUBLISHED THESE, IN THE HOPE 2790 02:02:57,680 --> 02:03:00,000 THAT PEOPLE WILL READ THEM AND 2791 02:03:00,000 --> 02:03:02,560 REALLY THINK ABOUT THEY'RE DOING 2792 02:03:02,560 --> 02:03:03,840 DPLI COSALATION REACTIONS AND WE 2793 02:03:03,840 --> 02:03:06,160 THINK IF YOU FOLLOW THIS LIST OF 2794 02:03:06,160 --> 02:03:07,200 GUIDELINES THAT THEN, WE SHOULD 2795 02:03:07,200 --> 02:03:10,800 BE ON THE WAY TO MAKING THIS 2796 02:03:10,800 --> 02:03:11,680 FIELD MORE REPRODUCIBLE, WHICH 2797 02:03:11,680 --> 02:03:14,320 WOULD BE AN ACHIEVEMENT, SO THE 2798 02:03:14,320 --> 02:03:16,280 QUESTION AND THE LAST SLIDE IS 2799 02:03:16,280 --> 02:03:18,240 ABOUT THE NEXT STEPS AND WHERE 2800 02:03:18,240 --> 02:03:21,760 DO WE GO FROM HERE. 2801 02:03:21,760 --> 02:03:24,000 AND WHAT I WOULD REALLY LIKE FOR 2802 02:03:24,000 --> 02:03:28,120 US TO DO NOT JUST MY LAB BUT AS 2803 02:03:28,120 --> 02:03:34,200 A COMMUNITY IS TO BUILD ON THE 2804 02:03:34,200 --> 02:03:38,640 GIED THAT WE PUBLISHED AND BUILD 2805 02:03:38,640 --> 02:03:40,320 STANDARDS ON THE DPLI 2806 02:03:40,320 --> 02:03:41,880 COSALATION'S ACTIONS SO THAT 2807 02:03:41,880 --> 02:03:42,840 PEOPLE CAN REPRODUCE THEM 2808 02:03:42,840 --> 02:03:44,880 THOUGHTFUL AND THEN FOR MAKING 2809 02:03:44,880 --> 02:03:46,080 COMPARISONS BETWEEN METHODS, SO 2810 02:03:46,080 --> 02:03:47,720 THAT WE CAN--WE CAN REALLY 2811 02:03:47,720 --> 02:03:49,880 DECIDE WHICH 1 IS BEST, AND 2812 02:03:49,880 --> 02:03:52,400 WHICH METHOD WE SHOULD ALL 2813 02:03:52,400 --> 02:03:52,720 FOLLOW. 2814 02:03:52,720 --> 02:03:55,080 SO, THOSE ARE THE KIND OF THINGS 2815 02:03:55,080 --> 02:03:57,600 THAT PETER AND I ARE INTERESTED 2816 02:03:57,600 --> 02:03:58,600 IN. 2817 02:03:58,600 --> 02:04:00,240 AND CLEARLY THAT'S A WORK IN 2818 02:04:00,240 --> 02:04:02,680 PROGRESS AND I'M LOOKING FORWARD 2819 02:04:02,680 --> 02:04:07,320 TO THE FEEDBACK FROM OUR 2820 02:04:07,320 --> 02:04:08,040 SYNTHETIC COLLEAGUES, HOPEFULLY 2821 02:04:08,040 --> 02:04:09,640 AT LUNCH TIME AS TO HOW WE MIGHT 2822 02:04:09,640 --> 02:04:12,840 GO FORWARD WITH THAT. 2823 02:04:12,840 --> 02:04:16,640 SO, THIS WORK HAS BEEN DONE IN 2824 02:04:16,640 --> 02:04:19,840 GEORGIA AND IN TO LEEING LIDO. 2825 02:04:19,840 --> 02:04:21,920 CLEARLY WE SHOULD THANK DAVID 2826 02:04:21,920 --> 02:04:25,840 WALT AND THE AUTHORS OF THE NRC 2827 02:04:25,840 --> 02:04:30,520 REPORT. 2828 02:04:30,520 --> 02:04:33,720 AGAIN, PAM MARINO AND THE GRANT 2829 02:04:33,720 --> 02:04:35,320 AND MANY COWORKERS OVER MANY 2830 02:04:35,320 --> 02:04:36,000 YEARS FOR THE CONTRIBUTION. 2831 02:04:36,000 --> 02:04:47,240 SO THANK YOU. 2832 02:04:47,240 --> 02:04:47,520 [ APPLAUSE ] 2833 02:04:47,520 --> 02:04:49,360 >> SO 1 THING TO BE AWARE OF IS 2834 02:04:49,360 --> 02:04:51,200 THAT FLOW CHEMIST RADIOY ALLOWS 2835 02:04:51,200 --> 02:04:52,960 TO YOU DO CONSTANT TEMPERATURE, 2836 02:04:52,960 --> 02:04:54,080 SO THAT'S WHY WE'RE EXCITED 2837 02:04:54,080 --> 02:04:56,240 ABOUT LOOKING AT THOSE OF THOSE 2838 02:04:56,240 --> 02:04:58,240 REACS LOOKING AT THESE 2839 02:04:58,240 --> 02:04:59,360 CONDITIONS, THIS ENDS THE EARLY 2840 02:04:59,360 --> 02:05:00,640 MORNING PROGRAM AND NOW I 2841 02:05:00,640 --> 02:05:07,160 TURNOVER THE FLOOR TO AMANDA. 2842 02:05:07,160 --> 02:05:09,560 I JUST WANT TO MAKE A FEW QUICK 2843 02:05:09,560 --> 02:05:10,640 ANNOUNCEMENTS, WE ARE GOING TO 2844 02:05:10,640 --> 02:05:12,320 SKIP THE BREAK, SO IF YOU NEED A 2845 02:05:12,320 --> 02:05:17,560 BREAK, LEAVE, COME BACK IN. 2846 02:05:17,560 --> 02:05:18,080 BLAWZ WE'RE RUNNING LATE. 2847 02:05:18,080 --> 02:05:20,320 WE WILL RUN INTO THE NEXT 2848 02:05:20,320 --> 02:05:21,600 SESSION CHAIRED BY DR. TMYER BUT 2849 02:05:21,600 --> 02:05:23,520 JUST A FEW OTHER ANNOUNCEMENTS 2850 02:05:23,520 --> 02:05:26,440 IF YOU EVAPORATE PICKED UP YOUR 2851 02:05:26,440 --> 02:05:27,640 BREAKFAST, PICK UP YOUR 2852 02:05:27,640 --> 02:05:29,920 BREAKFAST AND MAKURE YOU PAY PAM 2853 02:05:29,920 --> 02:05:31,440 FOR BREAKFAST, LUNCH AND DINNER 2854 02:05:31,440 --> 02:05:32,760 IF YOU ORDERED ANY OF THOSE AND 2855 02:05:32,760 --> 02:05:36,120 IF YOU ARE IN THE AFTERNOON 2856 02:05:36,120 --> 02:05:38,280 SESSION, AT 12:15 AS SOON AS 2857 02:05:38,280 --> 02:05:39,800 THIS SESSION ENDS PLEASE COME UP 2858 02:05:39,800 --> 02:05:40,720 AND UPLOAD YOUR SLIDE FIST YOU 2859 02:05:40,720 --> 02:06:02,080 EVAPORATE DONE SO YET. 2860 02:06:02,080 --> 02:06:04,280 THANK YOU. 2861 02:06:04,280 --> 02:06:06,240 >> --DR. CHEN, 60 SIS AND 2862 02:06:06,240 --> 02:06:25,400 REPLICATION OF GLYCO LIPIDS. 2863 02:06:25,400 --> 02:06:26,360 >> THANK YOU FOR THE 2864 02:06:26,360 --> 02:06:27,960 INTRODUCTION, AND I WOULD LIKE 2865 02:06:27,960 --> 02:06:29,440 TO THANK OTHER ORGANIZERS FOR 2866 02:06:29,440 --> 02:06:30,560 PUTTING THIS MEETING TOGETHER. 2867 02:06:30,560 --> 02:06:31,520 IT'S ALWAYS A GREAT PLEASURE TO 2868 02:06:31,520 --> 02:06:32,920 BE HERE IN PERSON. 2869 02:06:32,920 --> 02:06:37,920 AND TODAY, I AM GOING TO SHARE 2870 02:06:37,920 --> 02:06:39,560 WITH YOU OUR CONTRIBUTION FOR 2871 02:06:39,560 --> 02:06:48,000 THE DEVELOPMENT OF METHODS, 2872 02:06:48,000 --> 02:06:50,400 REGARDING WITH THE FOCUS ON 2873 02:06:50,400 --> 02:06:55,040 MEDICARE 2874 02:06:55,040 --> 02:06:56,080 MAMMALIAN LIPIDS. 2875 02:06:56,080 --> 02:06:57,840 HAD IS A COLLABORATION WITH MY 2876 02:06:57,840 --> 02:06:59,680 GROUP AT UC DAVIS AND THE 2877 02:06:59,680 --> 02:07:04,320 PROFESSOR GROUP AT THE GEORGIA 2878 02:07:04,320 --> 02:07:05,600 STATE UNIVERSITY. 2879 02:07:05,600 --> 02:07:08,040 AND THIS WAS A COLLABORATION 2880 02:07:08,040 --> 02:07:11,680 EFFORT THAT IS FUNDED BY UO1 2881 02:07:11,680 --> 02:07:17,040 COMMON FUND FLIEK O SCIENCE 2882 02:07:17,040 --> 02:07:17,240 PROGRAM. 2883 02:07:17,240 --> 02:07:21,360 ANDUREENTLY WITH WORKING WITH 2884 02:07:21,360 --> 02:07:24,160 IMCS IN TRANSLATING OUR P1S, THE 2885 02:07:24,160 --> 02:07:28,200 STRATEGIES AND THE KIDS FOR 2886 02:07:28,200 --> 02:07:35,840 ACCESSING FLIEK O SINGLE LIPPITY 2887 02:07:35,840 --> 02:08:00,160 EDUCATIONALS BY COMMERCIALIZING 2888 02:08:00,160 --> 02:08:01,240 THEM--HOWEVER, DETAILS OF THIS 2889 02:08:01,240 --> 02:08:03,720 MECHANISM IS NOT CLEAR. 2890 02:08:03,720 --> 02:08:07,600 THE MAJOR REASONS IS THE LACKING 2891 02:08:07,600 --> 02:08:10,560 OF FOR EXCESS TO STRUCTURALLY 2892 02:08:10,560 --> 02:08:14,400 DEFINED GLUE MARIOUS COSSINGLE 2893 02:08:14,400 --> 02:08:14,640 LIPIDS. 2894 02:08:14,640 --> 02:08:17,200 AND IN ADDITION TO SERVING 2895 02:08:17,200 --> 02:08:21,440 EXTENDERS, AND THE PROBES FOR 2896 02:08:21,440 --> 02:08:27,240 RESEARCH, LIPIDS ARE POTENTIAL 2897 02:08:27,240 --> 02:08:28,440 THERAPEUTIC REAGENTS ESPECIALLY 2898 02:08:28,440 --> 02:08:30,240 GREAT POTENTIAL FOR TREATING NEW 2899 02:08:30,240 --> 02:08:32,720 ROLE DAMAGES AND THE NEUROLOGY 2900 02:08:32,720 --> 02:08:35,040 CODE DISEASES SUCH AS HUNTING 2901 02:08:35,040 --> 02:08:37,040 TONS AND THE PARKINSON'S 2902 02:08:37,040 --> 02:08:39,680 DISEASES AND THERE'S A URGENT 2903 02:08:39,680 --> 02:08:43,400 NEED TO ACCESS THESE 2904 02:08:43,400 --> 02:08:44,280 STRUCTURALLY DEFINED GLYCO 2905 02:08:44,280 --> 02:08:45,760 SINGLE LIPIDS IF LARGE AMOUNT 2906 02:08:45,760 --> 02:08:47,840 ESPECIALLY IF YOU WANT TO USE 2907 02:08:47,840 --> 02:08:50,320 THEM AS A THERAPEUTIC REAGENTS 2908 02:08:50,320 --> 02:08:53,840 AND LET'S TAKE A LOOK AT THE 2909 02:08:53,840 --> 02:08:54,760 COMMERCIALLY AVAILABLE SOURCES. 2910 02:08:54,760 --> 02:08:58,240 ACTUALLY THOSE ARE VERY LIMITED. 2911 02:08:58,240 --> 02:09:01,040 AS YOU CAN SEE FROM THE 2912 02:09:01,040 --> 02:09:03,880 [INDISCERNIBLE] POLEAR LIPIDS, 2913 02:09:03,880 --> 02:09:06,400 THIS SIMPLE [INDISCERNIBLE] 2914 02:09:06,400 --> 02:09:08,960 DENDRITIC CELLROCYTE, THE SPRIES 2915 02:09:08,960 --> 02:09:10,000 EXTREMELY HIGH. 2916 02:09:10,000 --> 02:09:13,600 AND THE MORE PROBLEMATICALLY, 2917 02:09:13,600 --> 02:09:15,600 THE AVAILABLE REAGENT ACTUALLY 2918 02:09:15,600 --> 02:09:18,520 IS A MIXTURE AS THE POINT OUT IN 2919 02:09:18,520 --> 02:09:26,400 THE WEBSITE, THIS ONLY IS THE 1 2920 02:09:26,400 --> 02:09:28,800 OF THE STRUCTURES THAT SO AMINE 2921 02:09:28,800 --> 02:09:32,160 SYSTEM VERY HARD TO ACCESS 2922 02:09:32,160 --> 02:09:33,000 STRUCKURALLY DEFINED GLYCO 2923 02:09:33,000 --> 02:09:35,880 SINGLE LIPIDS WITH DEFINED GLYCO 2924 02:09:35,880 --> 02:09:41,640 COMPONENT AND THE LIPID 2925 02:09:41,640 --> 02:09:42,040 COMPONENT. 2926 02:09:42,040 --> 02:09:43,240 AND MAMMALIAN LIPIDS CAN BE 2927 02:09:43,240 --> 02:09:45,040 MOVED INTO 5 GROUPS. 2928 02:09:45,040 --> 02:09:49,160 ALL OF THEM ACTUALLY HAS THIS 2929 02:09:49,160 --> 02:09:49,800 COMMON [INDISCERNIBLE] ABILITY 2930 02:09:49,800 --> 02:09:59,000 COMPONENT WHICH IS CALLED ACET 2931 02:09:59,000 --> 02:09:59,240 MID. 2932 02:09:59,240 --> 02:10:00,440 THIS HAS SINGLE STRAND COMBONENT 2933 02:10:00,440 --> 02:10:01,880 WHICH IS FOUND WITH THE FATTY 2934 02:10:01,880 --> 02:10:04,760 ACID, BOTH OF THESE COMPONENTS 2935 02:10:04,760 --> 02:10:06,360 CAN BE DIFFERENT. 2936 02:10:06,360 --> 02:10:08,840 SO FOR THE SINGLE CELL PART, IN 2937 02:10:08,840 --> 02:10:12,240 NATURE YOU CAN 28 OR 18 2938 02:10:12,240 --> 02:10:12,640 [INDISCERNIBLE]. 2939 02:10:12,640 --> 02:10:14,720 FOLD UP THE ASSAY FOR THE 2940 02:10:14,720 --> 02:10:16,240 STRUCTURE OF VARIOUS PREMELY 2941 02:10:16,240 --> 02:10:20,520 CANNED WHAT DIFFERENT ON THE 2942 02:10:20,520 --> 02:10:22,640 SIZE AND ALSO DEGREE OF 2943 02:10:22,640 --> 02:10:27,000 SATURATION AND ALSO STRUCTURAL 2944 02:10:27,000 --> 02:10:27,320 MODIFICATIONS. 2945 02:10:27,320 --> 02:10:29,840 ALSO WITH THE LIPIDS ACTUALLY 2946 02:10:29,840 --> 02:10:32,440 SHARE A COMMON CORE WITH LACTOSE 2947 02:10:32,440 --> 02:10:35,960 THAT IS BETA TO THE CELL 2948 02:10:35,960 --> 02:10:36,320 [INDISCERNIBLE]. 2949 02:10:36,320 --> 02:10:38,280 AND THIS CELL BY ITSELF IS 2950 02:10:38,280 --> 02:10:40,440 INSOLUBLE IN WATER. 2951 02:10:40,440 --> 02:10:43,240 HOWEVER, IF YOU REMEEF THIS 2952 02:10:43,240 --> 02:10:43,440 APART. 2953 02:10:43,440 --> 02:10:46,920 THE LACTOSE SINGLE ASSAY ITSELF, 2954 02:10:46,920 --> 02:10:47,880 IT'S A RELATIVELY--READILY 2955 02:10:47,880 --> 02:10:51,240 SOLUBLE IN WATER, SO IT'S A VERY 2956 02:10:51,240 --> 02:10:53,840 SUITABLE ACCEPTOR SUBJECT FOR 2957 02:10:53,840 --> 02:10:55,040 GLUE MARIOUSICOSAL TRANSFER ACE 2958 02:10:55,040 --> 02:10:56,560 CATALYZE REACTION THAT CARRIED 2959 02:10:56,560 --> 02:10:58,240 OUT IN THE SOLUTION. 2960 02:10:58,240 --> 02:11:02,440 AND ALSO THE NATURAL OCCURRING 2961 02:11:02,440 --> 02:11:03,440 [INDISCERNIBLE] COMPONENT 2962 02:11:03,440 --> 02:11:06,000 PROVIDES A VERY GOOD TECH, 2963 02:11:06,000 --> 02:11:11,880 HYDROPHOBIC TECH TO ALIEU EITHER 2964 02:11:11,880 --> 02:11:13,280 PURIFICATION OF THE PRODUCTS, SO 2965 02:11:13,280 --> 02:11:16,640 OUR GOAL TO SET UP AT THE 2966 02:11:16,640 --> 02:11:21,280 BEGINNING OF THE YEAR WAS TO 2967 02:11:21,280 --> 02:11:22,640 DEVELOP PURIFICATION STRATEGIES 2968 02:11:22,640 --> 02:11:24,480 FOR FLIEK O SINGLE LIPIDS AND 2969 02:11:24,480 --> 02:11:27,280 TO,A LOW EVEN NONSPECIALISTS CAN 2970 02:11:27,280 --> 02:11:30,800 ACCESS THIS KIND OF COMPOUNDS 2971 02:11:30,800 --> 02:11:34,000 WITH VERY CONVENIENT TOOL BOX 2972 02:11:34,000 --> 02:11:39,840 AND ALSO VERY EASY PROCEDURES. 2973 02:11:39,840 --> 02:11:44,000 SO WE BASE THIS ON IMPORTANT 2974 02:11:44,000 --> 02:11:44,800 INTERMEDIATE LACTOSE SELLS WHERE 2975 02:11:44,800 --> 02:11:47,720 WE GIVE UP OUR STRATEGIES IN 3 2976 02:11:47,720 --> 02:11:48,040 STEPS. 2977 02:11:48,040 --> 02:11:51,160 THE FIRST STEP BASICALLY IS BY 2978 02:11:51,160 --> 02:11:53,200 CHEMICAL SYNTHESIS, TO ACCESS 2979 02:11:53,200 --> 02:11:55,840 MODULE AMOUNT OF LACTOSE SINGLE 2980 02:11:55,840 --> 02:11:59,120 STRAND AND THEN, STEP 2, IS TO 2981 02:11:59,120 --> 02:12:01,880 DO THE ENZYMATIC EXTENSION OF 2982 02:12:01,880 --> 02:12:06,360 THE GLYCO COMPONENT OF AND THIS 2983 02:12:06,360 --> 02:12:08,840 ACTUALLY IS THE BASED ON 2984 02:12:08,840 --> 02:12:11,240 THEENTIOUS FICIENCY OF THIS 2985 02:12:11,240 --> 02:12:13,640 GLYCO TRANSFER ACE CONTAINING 1 2986 02:12:13,640 --> 02:12:17,280 PARTICLE ENZYME SYSTEM, WITH THE 2987 02:12:17,280 --> 02:12:21,120 FACILITATION OF THE SINGLES IN 2988 02:12:21,120 --> 02:12:21,840 COMPONENT FOR PURIFICATION. 2989 02:12:21,840 --> 02:12:28,240 SOME WE HAVE OBTAINED THE GLYCO 2990 02:12:28,240 --> 02:12:30,400 [INDISCERNIBLE] INTERMEDIATES, 2991 02:12:30,400 --> 02:12:32,200 BY A SINGLE PURIFICATION IN LESS 2992 02:12:32,200 --> 02:12:34,920 THAN NIRT MINUTES AND STEP 3, 2993 02:12:34,920 --> 02:12:37,920 BASICALLY IS DOING THE ISOLATION 2994 02:12:37,920 --> 02:12:39,760 WHICH IS VERYENTIOUS FICIENT 2995 02:12:39,760 --> 02:12:42,240 STRATEGY TO INSTALL DIFFERENT 2996 02:12:42,240 --> 02:12:47,120 KIND OF FATTY TRAIN IN THE LAST 2997 02:12:47,120 --> 02:12:49,040 STEP TO PROVIDE A WIDE VARIETY 2998 02:12:49,040 --> 02:12:54,480 OF THE TARGET STRUCTURES. 2999 02:12:54,480 --> 02:12:56,320 SO STEP 1, ACTUALLY WEIGHT AND 3000 02:12:56,320 --> 02:12:59,240 GRABBED THE 4 GENERATIONS OF THE 3001 02:12:59,240 --> 02:13:00,640 SYNTHETIC STRATEGIES. 3002 02:13:00,640 --> 02:13:03,960 THE FIRST GENERATION WAS ALWAYS 3003 02:13:03,960 --> 02:13:05,440 THE SINGLE SCENE, WE HAVE 8 3004 02:13:05,440 --> 02:13:09,280 STEPS OF THE TRANSFORMATION, WE 3005 02:13:09,280 --> 02:13:13,080 OBTAINED THIS IMPORTANT TPHREU 3006 02:13:13,080 --> 02:13:14,440 RACING CO ACCEPTOR THAT ALLOW US 3007 02:13:14,440 --> 02:13:16,920 TO DO THE DPLI COSALATION 3008 02:13:16,920 --> 02:13:21,040 REACTION WITH LACTOSE DERIVED 3009 02:13:21,040 --> 02:13:21,240 DONOR. 3010 02:13:21,240 --> 02:13:24,240 WE WERE AICIAL TO DO THIS 3011 02:13:24,240 --> 02:13:26,280 REACTION IN LARGER SKILL AND 3012 02:13:26,280 --> 02:13:29,840 WITH RELATIVELY HIGH YIELDS. 3013 02:13:29,840 --> 02:13:31,600 THE LIMITATION ACTUALLY IS THAT 3014 02:13:31,600 --> 02:13:35,640 WE CAN ONLY OBTAIN THE LACTIC 3015 02:13:35,640 --> 02:13:38,840 ACIDOSEISS ON [INDISCERNIBLE] 3016 02:13:38,840 --> 02:13:41,480 AND THE OTHER SPINNING O SWEEN, 3017 02:13:41,480 --> 02:13:46,840 WITH THE 18 COVERS SPHINGOSIGN, 3018 02:13:46,840 --> 02:13:51,160 AND THIS START WITH THE LESS 3019 02:13:51,160 --> 02:13:52,680 EXPENSIVE L-SERIES POINTSINE 3020 02:13:52,680 --> 02:13:53,160 BUILDING BLOCK. 3021 02:13:53,160 --> 02:13:56,840 AND WE'RE ABLE TO ACHIEVE ALSO 3022 02:13:56,840 --> 02:14:07,600 MORE THAN 10 GRAND SCALE SYNTH 3023 02:14:07,600 --> 02:14:10,040 AND THE FOURTH GENERATION OF THE 3024 02:14:10,040 --> 02:14:11,560 SYNTHETIC STEPS AND THIS 3025 02:14:11,560 --> 02:14:14,000 ACTUALLY ALLOWS TO ACCESS THE 3026 02:14:14,000 --> 02:14:16,280 TARGET IN LESS STEPS AND THE 3027 02:14:16,280 --> 02:14:19,120 ALSO WITH MUCH LOWER COST AND 3028 02:14:19,120 --> 02:14:24,320 THIS WILL ALLOW US TO BE ABLE TO 3029 02:14:24,320 --> 02:14:28,240 PRODUCE THE LACTOSE SPINGOSIGN 3030 02:14:28,240 --> 02:14:30,880 IN LESS EXPENSIVE COST. 3031 02:14:30,880 --> 02:14:50,520 SO WITH THE LACTOSE AND--AND AND 3032 02:14:50,520 --> 02:14:55,440 THE DIVERSE OF THE TARGETS. 3033 02:14:55,440 --> 02:14:58,160 AND EVERY STEP ACTUALLY SAY 3034 02:14:58,160 --> 02:15:00,200 PURIFICATION ALLOW US TO GET THE 3035 02:15:00,200 --> 02:15:02,840 TARGET IN LESS THAN 30 MINUTES. 3036 02:15:02,840 --> 02:15:08,840 AND I ALSO WANT TO HIGHLIGHT THE 3037 02:15:08,840 --> 02:15:10,280 ADVANTAGE OF THE ENZYME SYSTEM 3038 02:15:10,280 --> 02:15:12,640 ALLOWS US TO INTRODUCE MODIFIED 3039 02:15:12,640 --> 02:15:17,560 SUGAR STRUCTURES, INTO THE 3040 02:15:17,560 --> 02:15:18,880 TARGETS. 3041 02:15:18,880 --> 02:15:20,840 SO OVERALL 15, 1 PART 3042 02:15:20,840 --> 02:15:23,240 [INDISCERNIBLE] SYSTEMS CAN BE 3043 02:15:23,240 --> 02:15:26,120 USED TO GENERATE MOST OF THE 3044 02:15:26,120 --> 02:15:27,520 GLYCO LIPID TARGETS AND I WILL 3045 02:15:27,520 --> 02:15:31,440 HIGHLIGHT 4 OF THEM WHICH CAN BE 3046 02:15:31,440 --> 02:15:35,040 USED TO SYNTHESIZE MAJORITY OF 3047 02:15:35,040 --> 02:15:37,600 THE IMPORTANT CELLULAR GLYCO 3048 02:15:37,600 --> 02:15:38,840 [INDISCERNIBLE]. 3049 02:15:38,840 --> 02:15:41,800 AND OF THEM, ACTUALLY START WITH 3050 02:15:41,800 --> 02:15:45,240 SIMPLE MONOSACCHARIDES WHICH ARE 3051 02:15:45,240 --> 02:15:45,840 INEXPENSIVE AND COMMERCIALLY 3052 02:15:45,840 --> 02:15:49,240 AVAILABLE AND THEY CAN BE 3053 02:15:49,240 --> 02:15:50,560 ACTIVATED TO GENERATE SUGAR 3054 02:15:50,560 --> 02:15:53,000 NUCLEOTIDE THAT CAN BE USED UP 3055 02:15:53,000 --> 02:15:55,480 BY THE SUITABLE TRANSFER ACE TO 3056 02:15:55,480 --> 02:15:58,400 MAKE THE THESE OTHER TARGETS. 3057 02:15:58,400 --> 02:16:01,440 AGAIN THE COMPONENT IN THE 3058 02:16:01,440 --> 02:16:03,960 PRODUCT HELPS FOR THE 3059 02:16:03,960 --> 02:16:06,240 PURIFICATIONS PROCESS THAT CAN 3060 02:16:06,240 --> 02:16:09,960 BE DONE IN LESS THAN 30 MINUTES, 3061 02:16:09,960 --> 02:16:12,440 USE THE SYMPOSIUM 3062 02:16:12,440 --> 02:16:14,040 [INDISCERNIBLE]. 3063 02:16:14,040 --> 02:16:19,840 AND AS A THUMB POST HAS 3064 02:16:19,840 --> 02:16:21,600 SUCCESSFULLY SYNTHESIZED FOR 3065 02:16:21,600 --> 02:16:28,760 GANGLIA BASED PRIORITIZED CANCER 3066 02:16:28,760 --> 02:16:30,600 AND THE GENES WERE COMPLETED 3067 02:16:30,600 --> 02:16:33,400 WITH 1 OR MORE REACTIONS WITH 3068 02:16:33,400 --> 02:16:37,440 VARIOUS SKILLS AND FOR THE 3069 02:16:37,440 --> 02:16:38,080 [INDISCERNIBLE], THE INDIVIDUAL 3070 02:16:38,080 --> 02:16:42,480 TO MAKE MORE THAN 1 GRAM IN THIS 3071 02:16:42,480 --> 02:16:46,160 PRODUCT AND AFTER STEPS ACTUALLY 3072 02:16:46,160 --> 02:16:48,840 OBTAINED IN HIGH YIELDS, 3073 02:16:48,840 --> 02:16:50,960 APPROXIMATE THE USE ISOLATION 3074 02:16:50,960 --> 02:16:54,800 CONDITION THAT PURIFY THE 3075 02:16:54,800 --> 02:16:56,640 PRODUCT VERY EFFICIENTLY. 3076 02:16:56,640 --> 02:16:58,280 AND WE ALSO ALSO INTRODUCE 3077 02:16:58,280 --> 02:17:01,240 MODIFY SUGARS TO THE FINAL 3078 02:17:01,240 --> 02:17:05,320 PRODUCT AS A DRUM HOLE, THERE'S 3079 02:17:05,320 --> 02:17:08,160 ALWAYS THIS [INDISCERNIBLE] 3080 02:17:08,160 --> 02:17:11,200 ASSAY, AND USE SALIC ACID, 3081 02:17:11,200 --> 02:17:15,960 OTHERWISE BASE THE REACTION TO 3082 02:17:15,960 --> 02:17:18,320 GENERATE THIS GM-3 CONTAINING 3083 02:17:18,320 --> 02:17:24,600 DIFFERENT SIALOGLYCANSIC ACID 3084 02:17:24,600 --> 02:17:24,800 FORMS. 3085 02:17:24,800 --> 02:17:25,760 AND CURRENTLY WE ARE WORKING 3086 02:17:25,760 --> 02:17:29,400 WITH THIS TO FURTHER IMPROVE THE 3087 02:17:29,400 --> 02:17:31,120 STABILITY, EXPRESSION LEVELS OF 3088 02:17:31,120 --> 02:17:33,160 THE KEY FLIEK O TRANSFER ACES 3089 02:17:33,160 --> 02:17:36,520 THAT ARE INVOLVED IN THIS 3090 02:17:36,520 --> 02:17:39,440 SYNTHETIC PROCESS, BY APROTEIN 3091 02:17:39,440 --> 02:17:43,640 ENGINEERING PROCESS AND ALSO 3092 02:17:43,640 --> 02:17:44,840 WITH IMCS EXPERTS. 3093 02:17:44,840 --> 02:17:46,760 ON THE FRAGMENTATION PROCEDURES. 3094 02:17:46,760 --> 02:17:51,040 AND WE ALSO FURTHER IMPROVE THE 3095 02:17:51,040 --> 02:17:53,120 PROCESS, BY IMPROVING CHEMICAL 3096 02:17:53,120 --> 02:17:55,760 SYNTHESIS OF THE LACTOSE 3097 02:17:55,760 --> 02:17:57,640 [INDISCERNIBLE] AS WELL AS TO 3098 02:17:57,640 --> 02:17:59,160 IMPROVE THE OPMI AND THE MEDICAL 3099 02:17:59,160 --> 02:18:02,040 PROCESS, IF WE HAVE OUR TARGET 3100 02:18:02,040 --> 02:18:04,680 IN MIND, ACTUALLY WE DON'T NEED 3101 02:18:04,680 --> 02:18:08,440 TO PURIFY THE INTERMEDIATE 3102 02:18:08,440 --> 02:18:11,640 PRODUCTS AND BY THIS WAY, WE CAN 3103 02:18:11,640 --> 02:18:13,640 MINIMIZE THE PURIFICATIONS STEPS 3104 02:18:13,640 --> 02:18:16,120 AND ALSO WE FURTHER IMPROVE THE 3105 02:18:16,120 --> 02:18:18,040 PROCESS BY CHANGING THE 3106 02:18:18,040 --> 02:18:20,600 CONDITION AND THE DECREASED 3107 02:18:20,600 --> 02:18:22,240 AMOUNT OF THE ENZYME THAT CAN BE 3108 02:18:22,240 --> 02:18:24,560 USED AND WITH SUCCESS RATE, WE 3109 02:18:24,560 --> 02:18:26,680 DID THIS, GRAM SKILL SYNTHESIS 3110 02:18:26,680 --> 02:18:30,880 OF THE GM 1 AND WE ARE WORKING 3111 02:18:30,880 --> 02:18:31,640 WITH MATERIALS MAKING THE 3112 02:18:31,640 --> 02:18:33,160 10-GRAM SCALE AND MAYBE EVEN 3113 02:18:33,160 --> 02:18:36,280 LARGER SCALE OF THAT AND WITH 3114 02:18:36,280 --> 02:18:37,720 THOSE TARGET FLIEK O 3115 02:18:37,720 --> 02:18:40,200 [INDISCERNIBLE] IN HAND WE CAN 3116 02:18:40,200 --> 02:18:42,080 DO THE FINAL ISOLATION REACTION 3117 02:18:42,080 --> 02:18:49,040 AND A COMPELLED TO OUR EARLIER 3118 02:18:49,040 --> 02:18:51,000 CONDITIONS WHICH WASSA, CHIEFED 3119 02:18:51,000 --> 02:18:52,440 25% YIELD IN 84 HOURS STARTING 3120 02:18:52,440 --> 02:18:55,680 WITH THE FATTY ASSAY AS 1 OF THE 3121 02:18:55,680 --> 02:18:56,040 STUDY MATERIAL. 3122 02:18:56,040 --> 02:18:58,400 WE FOUND OUT THE ACL 3123 02:18:58,400 --> 02:18:59,640 [INDISCERNIBLE] IS A BETTER 3124 02:18:59,640 --> 02:19:01,560 STUDY MATERIAL AND THE REACTION 3125 02:19:01,560 --> 02:19:06,880 CAN BE COMPLETED IN 2 HOURS 3126 02:19:06,880 --> 02:19:09,080 IEWOF THE JUSTUTESSING THE 3127 02:19:09,080 --> 02:19:09,960 COGNATE IN [INDISCERNIBLE]. 3128 02:19:09,960 --> 02:19:11,960 AND WE HAVE VERY HIGH YIELDS FOR 3129 02:19:11,960 --> 02:19:15,040 THE TARGETS AND AS EXAMPLE, WE 3130 02:19:15,040 --> 02:19:17,840 CAN STORE DIFFERENT SIZES. 3131 02:19:17,840 --> 02:19:19,160 AND DIFFERENT DEGREE OF 3132 02:19:19,160 --> 02:19:21,120 SATTURATION OF THE FATTY ACID 3133 02:19:21,120 --> 02:19:24,360 SAY ON TO THE FINAL PRODUCT. 3134 02:19:24,360 --> 02:19:26,120 SO BASICALLY ALL THIS WERE 3135 02:19:26,120 --> 02:19:30,440 ACHIEVED IN VERY HIGH YIELDS. 3136 02:19:30,440 --> 02:19:35,600 SO IN SUMMARY, WE HAVE ACHIEVED 3137 02:19:35,600 --> 02:19:36,840 OUR NUMBER 1 GOAL AND OUR RESULT 3138 02:19:36,840 --> 02:19:39,480 VS BEEN SHARED WITH THE 3139 02:19:39,480 --> 02:19:42,040 COMMUNITY BY RESEARCH PAPERS, WE 3140 02:19:42,040 --> 02:19:44,720 ARE ALSO STILL IN THE PROCESS OF 3141 02:19:44,720 --> 02:19:49,240 REPAIRING NEW PAPERS WHICH WILL 3142 02:19:49,240 --> 02:19:50,880 COME OUT LATER IN YEAR AND THE 3143 02:19:50,880 --> 02:19:53,400 PROTOCOL FOR THE SIPGHT SIS OF 3144 02:19:53,400 --> 02:19:58,960 THE GD3 AND ALSO GM3, COVERAGED 3145 02:19:58,960 --> 02:20:01,840 IN THIS GLYCO SCIENCE SPECIAL 3146 02:20:01,840 --> 02:20:03,320 COLLECTION ORGANIZED BY 3147 02:20:03,320 --> 02:20:06,240 [INDISCERNIBLE] AND ALSO 3148 02:20:06,240 --> 02:20:06,640 [INDISCERNIBLE]. 3149 02:20:06,640 --> 02:20:18,800 AND WE ALSO SHOW OUR RESULTS IN 3150 02:20:18,800 --> 02:20:20,040 TALKS AND THE THE CROSS 3151 02:20:20,040 --> 02:20:21,360 VALIDATION WAS DONE BY THE TEAM. 3152 02:20:21,360 --> 02:20:25,160 AND WE ALSO SHARE OUR ENZYMES 3153 02:20:25,160 --> 02:20:27,120 WITH SEVERAL GROUPS AND FOR MORE 3154 02:20:27,120 --> 02:20:27,720 DETAILED INFORMATION PLEASE 3155 02:20:27,720 --> 02:20:31,800 CHECK OUT THE WEBSITE THAT 3156 02:20:31,800 --> 02:20:35,840 DR. [INDISCERNIBLE] AT GEORGIA 3157 02:20:35,840 --> 02:20:36,440 STATE UNIVERSITY ESTABLISHED. 3158 02:20:36,440 --> 02:20:39,520 AND WITH THAT I WOULD LIKE TO 3159 02:20:39,520 --> 02:20:41,600 THANK OUR TEAM MEMBERS WHO 3160 02:20:41,600 --> 02:20:43,120 CONTRIBUTE TO THE WORK THAT ARE 3161 02:20:43,120 --> 02:20:46,240 PRESENTING AND ALSO OUR 3162 02:20:46,240 --> 02:20:49,400 COLLABORATORS AND THE GROUP FOR 3163 02:20:49,400 --> 02:20:50,320 CROSS VALIDATION EFFORT AND OF 3164 02:20:50,320 --> 02:20:54,560 COURSE I WOULD LIKE TO THANK THE 3165 02:20:54,560 --> 02:20:56,880 FINANCIAL SUPPORT FROM NIH 3166 02:20:56,880 --> 02:20:58,280 COMMON FUND GLYCO SCIENCE 3167 02:20:58,280 --> 02:21:04,480 PROGRAM AND ALSO NIGMS WITHOUT 3168 02:21:04,480 --> 02:21:06,440 THIS FINANCIAL SUPPORT, OUR 3169 02:21:06,440 --> 02:21:07,840 EFFORTS IN ADVANCING THE GLYCO 3170 02:21:07,840 --> 02:21:09,680 SCIENCE FIELD WILL NOT BE 3171 02:21:09,680 --> 02:21:10,040 POSSIBLE. 3172 02:21:10,040 --> 02:21:12,160 AND WITH THAT, IRB WOULD LIKE TO 3173 02:21:12,160 --> 02:21:19,920 THANK YOU FOR YOUR ATTENTION. 3174 02:21:19,920 --> 02:21:27,920 [ APPLAUSE ] 3175 02:21:27,920 --> 02:21:28,720 , THANKS. 3176 02:21:28,720 --> 02:21:32,680 >> OUR NEXT TALK IS FROM DR. YU 3177 02:21:32,680 --> 02:21:35,000 FROM UNIVERSITY CALIFORNIA DAVIS 3178 02:21:35,000 --> 02:21:37,680 AS WE RICOCHET BACK AND FORTH 3179 02:21:37,680 --> 02:21:40,880 BETWEEN POLYSACCHARIDES AND 3180 02:21:40,880 --> 02:21:41,800 LIPIDS. 3181 02:21:41,800 --> 02:21:46,120 FOR CHEMOEN ENZYMATIC SYNTHESIS 3182 02:21:46,120 --> 02:21:49,200 OF BACTERIAL POLYSACCHARIDES. 3183 02:21:49,200 --> 02:21:51,640 >> THANK YOU MICHAEL. 3184 02:21:51,640 --> 02:21:54,760 SO OUR U01 PROJECT IS ABOUT 3185 02:21:54,760 --> 02:21:56,400 CHEMO ENSWRANSS OF 3186 02:21:56,400 --> 02:21:56,760 POLYSACCHARIDE. 3187 02:21:56,760 --> 02:21:58,760 WE ARE CLOBERATING WITH LAURA AT 3188 02:21:58,760 --> 02:22:01,840 M. I.T. AND [INDISCERNIBLE] AT 3189 02:22:01,840 --> 02:22:04,240 GEORGIA STATE. 3190 02:22:04,240 --> 02:22:10,120 THE INTEREST CURRENTLY SVR, WITH 3191 02:22:10,120 --> 02:22:15,560 THE [INDISCERNIBLE] AND 3192 02:22:15,560 --> 02:22:22,840 COLLABORATE WITH [INDISCERNIBLE] 3193 02:22:22,840 --> 02:22:23,840 FOCUSING GENES SIPGHTICIZING 3194 02:22:23,840 --> 02:22:24,240 DPLI KAN--KANAS. 3195 02:22:24,240 --> 02:22:31,400 WE KNOW THIS IS A MAJOR PROBLEM, 3196 02:22:31,400 --> 02:22:35,000 BY THE SURVASE, IT'S A PACTERIAL 3197 02:22:35,000 --> 02:22:35,800 POLYSACCHARIDE. 3198 02:22:35,800 --> 02:22:37,880 SO THE POLYSACCHARIDE ON THE 3199 02:22:37,880 --> 02:22:39,240 BACTERIAL VECTOR, THE OTHER 3200 02:22:39,240 --> 02:22:44,680 VACCINE TARGET AND THE CAUSE OF 3201 02:22:44,680 --> 02:22:47,160 THE TYROSINE CANDIDATE, SO 3202 02:22:47,160 --> 02:22:49,440 DETECTED BY THE POLYSACCHARIDE 3203 02:22:49,440 --> 02:22:50,920 IS VERY POTENT. 3204 02:22:50,920 --> 02:22:54,240 SO THE PROBE TO STED THE RELATED 3205 02:22:54,240 --> 02:22:55,880 INTER--PROBE TO STUDY THE 3206 02:22:55,880 --> 02:22:57,760 RELATED INTERACTIONS AND THE 3207 02:22:57,760 --> 02:23:01,840 MATERIALS AND IMPROVE THE 3208 02:23:01,840 --> 02:23:02,320 PROTEIN THEOR PUTTICS. 3209 02:23:02,320 --> 02:23:03,840 SO ALSO AWARE OF THE IMPORTANT 3210 02:23:03,840 --> 02:23:09,040 PROBE TO STUDY THE ARBO HYDRATE 3211 02:23:09,040 --> 02:23:10,640 ENZYME HOWEVER CURRENTLY MANY OF 3212 02:23:10,640 --> 02:23:14,960 THE LABS SOMEHOW ARE AVAILABLE. 3213 02:23:14,960 --> 02:23:19,720 THE 3 TO PRODUCING 3214 02:23:19,720 --> 02:23:21,120 POLYSACCHARIDE IN THIS GROUP, 3215 02:23:21,120 --> 02:23:24,920 MANY ARE FOCUSING ON THE 3216 02:23:24,920 --> 02:23:28,200 SYNTHESIZING [INDISCERNIBLE] 3217 02:23:28,200 --> 02:23:29,560 DEPENDENT POLYSACCHARIDE TARGET. 3218 02:23:29,560 --> 02:23:34,080 LAURA IN OUR GROUP TRYING TO 3219 02:23:34,080 --> 02:23:36,360 [INDISCERNIBLE] INDEPENDENT 3220 02:23:36,360 --> 02:23:39,400 TARGET. 3221 02:23:39,400 --> 02:23:41,360 HOWEVER, WE PARTICULARLY 3222 02:23:41,360 --> 02:23:43,920 [INDISCERNIBLE] PALY SACCHARIDE, 3223 02:23:43,920 --> 02:23:50,600 INCLUDING THE [INDISCERNIBLE], 3224 02:23:50,600 --> 02:23:54,520 DEFINE, AND [INDISCERNIBLE]. 3225 02:23:54,520 --> 02:23:57,280 BACTERIA CAN CAUSE LIFE 3226 02:23:57,280 --> 02:24:00,000 [INDISCERNIBLE] MOTIFS. 3227 02:24:00,000 --> 02:24:02,640 SO THE BACTERIAL POINT HAS A 3228 02:24:02,640 --> 02:24:08,040 SEPARATE UNIT, WHICH IS BY THE 3229 02:24:08,040 --> 02:24:09,560 SYNTHESIS. 3230 02:24:09,560 --> 02:24:11,280 THE PROBLEM WITH THE SYNTHESIS 3231 02:24:11,280 --> 02:24:13,160 IS THAT THEY HAVE THE 3232 02:24:13,160 --> 02:24:15,160 [INDISCERNIBLE] PENN STATE FOR 3233 02:24:15,160 --> 02:24:17,400 THE IMPLAY 1 PORT TO THE ENZYME 3234 02:24:17,400 --> 02:24:20,200 FOR THE DPLI COSALATION COMBINED 3235 02:24:20,200 --> 02:24:23,720 WITH THE BACTERIAL 60 SIS TO 3236 02:24:23,720 --> 02:24:28,760 PRODUCE DPLI KAN--KANAS, IN THE 3237 02:24:28,760 --> 02:24:31,360 [INDISCERNIBLE] NUCLEOTIDE, AND 3238 02:24:31,360 --> 02:24:35,040 FROM THE SIMPLE MONOSACCHARIDE, 3239 02:24:35,040 --> 02:24:38,320 TRYING TO BY THE BACTERIAL 3240 02:24:38,320 --> 02:24:42,640 SYNTHESIS,OT 1 PAGE THE OPM E IS 3241 02:24:42,640 --> 02:24:45,760 AWARD USING [INDISCERNIBLE], 3242 02:24:45,760 --> 02:24:47,960 NUCLEOTIDE, AND THE NEED IT TO 3243 02:24:47,960 --> 02:24:48,800 INCORNERATE THE MODIFIED SUGAR 3244 02:24:48,800 --> 02:24:51,240 TO THE DPLI KAN--KANA SUCH AS 3245 02:24:51,240 --> 02:24:52,200 [INDISCERNIBLE] AND IKAN--KANA 3246 02:24:52,200 --> 02:24:59,600 GROUP TO ALLOW FOR THE 3247 02:24:59,600 --> 02:24:59,920 MODIFICATIONS. 3248 02:24:59,920 --> 02:25:05,080 SO BY EXAMPLE, THE OPME, WILL 3249 02:25:05,080 --> 02:25:07,320 ASSIMMULATE IN AN DROJ NOWS 3250 02:25:07,320 --> 02:25:13,480 [INDISCERNIBLE] POLYSACCHARIDE 3251 02:25:13,480 --> 02:25:17,160 DPLI KAN--KANA. 3252 02:25:17,160 --> 02:25:17,760 THE DISCIPLINARY SACCHARIDE, 3253 02:25:17,760 --> 02:25:20,040 SINCE THIS IS CALLED 3254 02:25:20,040 --> 02:25:21,640 [INDISCERNIBLE] BETTER FUNCTION 3255 02:25:21,640 --> 02:25:23,840 FOR THE ENZYME, YOU JUST KILLED 3256 02:25:23,840 --> 02:25:25,040 THE POLYSACCHARIDE AND YOU 3257 02:25:25,040 --> 02:25:26,240 UPROOT WE START ON THE 3258 02:25:26,240 --> 02:25:30,080 [INDISCERNIBLE] FOR THE NEW OOH 3259 02:25:30,080 --> 02:25:33,520 C, AND AND DPLI COSALATION IN 3260 02:25:33,520 --> 02:25:38,760 THE [INDISCERNIBLE] TO FORM THE 3261 02:25:38,760 --> 02:25:39,120 [INDISCERNIBLE]. 3262 02:25:39,120 --> 02:25:41,640 CAN SHOW THE GRAND SCALE 3263 02:25:41,640 --> 02:25:41,920 INCIDENT. 3264 02:25:41,920 --> 02:25:47,160 THE AFULLATION FROM THE 3265 02:25:47,160 --> 02:25:47,520 [INDISCERNIBLE]. 3266 02:25:47,520 --> 02:25:50,640 THE CPT LAB TO PURIFY THE 3267 02:25:50,640 --> 02:26:10,400 PRODUCT, USING THE IT CAN BE 3268 02:26:10,400 --> 02:26:11,720 EASILY REMOVED--GLYCOSYLATION, 3269 02:26:11,720 --> 02:26:16,920 IT'S A DIFFERENT SITE TO THE 3270 02:26:16,920 --> 02:26:17,240 SACCHARIDE. 3271 02:26:17,240 --> 02:26:22,680 UPON IT WAS ABLE TO STUDY THE 3272 02:26:22,680 --> 02:26:24,240 DIFFERENT SHAPE BETWEEN THE SET 3273 02:26:24,240 --> 02:26:29,400 OF THE SUBSET, AND THE 3274 02:26:29,400 --> 02:26:29,760 [INDISCERNIBLE]. 3275 02:26:29,760 --> 02:26:33,360 IT FUNDED THE GUARANTEE OF THE 3276 02:26:33,360 --> 02:26:40,000 [INDISCERNIBLE] WHEN YOU CAN 3277 02:26:40,000 --> 02:26:42,040 [INDISCERNIBLE] THE CENTER WILL 3278 02:26:42,040 --> 02:26:46,040 NOT AFFECT THE DOSE 3279 02:26:46,040 --> 02:26:46,400 [INDISCERNIBLE]. 3280 02:26:46,400 --> 02:26:49,760 SO FURTHER, THE [INDISCERNIBLE] 3281 02:26:49,760 --> 02:26:53,280 OF THIS [INDISCERNIBLE] ENZYME 3282 02:26:53,280 --> 02:26:54,680 VERY BROAD CONOR SPECIFICITY BUT 3283 02:26:54,680 --> 02:26:57,360 IT CAN TOLERATE A LOT OF 3284 02:26:57,360 --> 02:26:59,280 MODIFICATIONS ON THIS, 3285 02:26:59,280 --> 02:27:03,240 ESPECIALLY THE IT CAN TAKE THE 3286 02:27:03,240 --> 02:27:07,800 [INDISCERNIBLE], AND THE CMP, 3287 02:27:07,800 --> 02:27:13,520 ACID TOLERANT, THE PROPERTIES 3288 02:27:13,520 --> 02:27:15,280 USING THE [INDISCERNIBLE] OR THE 3289 02:27:15,280 --> 02:27:21,360 SIXTH [INDISCERNIBLE]. 3290 02:27:21,360 --> 02:27:22,840 ADDING SALIC ACID SEPRESSOR FROM 3291 02:27:22,840 --> 02:27:26,040 STARTING FROM THE SACCHARIDE,A 3292 02:27:26,040 --> 02:27:27,960 PLIES OPME, STARTING THE CANNED 3293 02:27:27,960 --> 02:27:29,440 WHAT AND RADIATION PRODUCE 3294 02:27:29,440 --> 02:27:32,160 LIBRARY FROM THE TRI SACCHARIDE, 3295 02:27:32,160 --> 02:27:34,320 TO UNDER THE 10 TO THE 3296 02:27:34,320 --> 02:27:36,280 SACCHARIDE, AND WHICH YOU SHOULD 3297 02:27:36,280 --> 02:27:38,360 THE STRICT ASSAY, THOSE ARE 3298 02:27:38,360 --> 02:27:40,240 SEVENTH OF THE NINTH 3299 02:27:40,240 --> 02:27:44,960 MODIFICATION, SO, IT CAN BE 3300 02:27:44,960 --> 02:27:47,240 READILY [INDISCERNIBLE] AS A 3301 02:27:47,240 --> 02:27:49,040 GROUP SO THOSE TPHREU RACING 3302 02:27:49,040 --> 02:27:51,760 KAN--KANA FLOWED AS A STABLE 3303 02:27:51,760 --> 02:28:02,800 PROBE AS THE TPHREU RACING 3304 02:28:02,800 --> 02:28:04,160 KAN--KANAS. 3305 02:28:04,160 --> 02:28:07,400 HAVE D AND THE K5 POLYSACCHARIDE 3306 02:28:07,400 --> 02:28:09,280 AND THE 2 BART ARE THE SAME, 3307 02:28:09,280 --> 02:28:11,840 THEY SEPARATE THE UNIT AND THE 3308 02:28:11,840 --> 02:28:14,120 PM IT SHOWED 2 IS THE BI 3309 02:28:14,120 --> 02:28:16,840 FUNCTIONAL ENZYME FOR THE YOUTH 3310 02:28:16,840 --> 02:28:18,840 TO SYNTHESIZE THESE 3311 02:28:18,840 --> 02:28:19,200 POLYSACCHARIDE. 3312 02:28:19,200 --> 02:28:23,120 AND ON THE TRUNCATED DATE, 3313 02:28:23,120 --> 02:28:24,720 8:00 P.M. TO 2 TO HOW TO IMPROVE 3314 02:28:24,720 --> 02:28:31,720 THE SUPPRESS AND HOW BETTER THE 3315 02:28:31,720 --> 02:28:36,680 STABILITY IS TRUNCATED IN THE 3316 02:28:36,680 --> 02:28:37,040 SENTENCES. 3317 02:28:37,040 --> 02:28:40,200 SO APPLYING THE OPME, AND THE 3318 02:28:40,200 --> 02:28:45,040 LOCAL TRANSFERAISES, START FROM 3319 02:28:45,040 --> 02:28:46,960 THE SMALL MONOSACCHARIDE WE 3320 02:28:46,960 --> 02:28:48,640 SYNTHESIZED UP TO 3321 02:28:48,640 --> 02:28:53,280 [INDISCERNIBLE] SACCHARIDE, SO 3322 02:28:53,280 --> 02:28:54,280 EACH PROGRAM WAS PRODUCED ON 3323 02:28:54,280 --> 02:28:54,680 SCALE. 3324 02:28:54,680 --> 02:28:56,680 SO WHEN TRYING SETTING, THE 3325 02:28:56,680 --> 02:28:59,160 GLYCAN SUCH AS A PHARMA 3326 02:28:59,160 --> 02:29:01,600 [INDISCERNIBLE] TO HAVE THAT 3327 02:29:01,600 --> 02:29:03,400 SACCHARIDE, FOR THE FIRST 3328 02:29:03,400 --> 02:29:06,160 CAUSEIDATION FOR THE ENZYME WAS 3329 02:29:06,160 --> 02:29:10,040 THERE, WHICH MEANS WHEN YOU TRY 3330 02:29:10,040 --> 02:29:14,600 TO USING THE SYNTHESIZE, BESIDES 3331 02:29:14,600 --> 02:29:17,440 THE HYDROCELLS ARE THE DIFFERENT 3332 02:29:17,440 --> 02:29:19,720 SET OF [INDISCERNIBLE] 3333 02:29:19,720 --> 02:29:21,640 [INDISCERNIBLE] TO OVERCOME THIS 3334 02:29:21,640 --> 02:29:24,000 [INDISCERNIBLE] WITH THE 2 3335 02:29:24,000 --> 02:29:24,240 MUTANTS. 3336 02:29:24,240 --> 02:29:26,640 ONE MUTANT, 1 SINGLE FUNCTION OF 3337 02:29:26,640 --> 02:29:32,240 ATT, AND HOW IT LOOKS NEGATIVE 2 3338 02:29:32,240 --> 02:29:32,520 TWORKS T. 3339 02:29:32,520 --> 02:29:34,160 AND TO TURN THIS OUT, THERE ARE 3340 02:29:34,160 --> 02:29:36,240 DPLI KAN--KANAS SO WE WERE ABLE 3341 02:29:36,240 --> 02:29:41,040 TO SEND THIS IN, OUT OF THE 3342 02:29:41,040 --> 02:29:53,640 SACCHARIDE, WITH THE YIELD. 3343 02:29:53,640 --> 02:30:01,360 OKAY, [INDISCERNIBLE] AND THE 3344 02:30:01,360 --> 02:30:04,000 CHEMO [INDISCERNIBLE] YOU START 3345 02:30:04,000 --> 02:30:11,480 FROM THE UDP SERUM WITH INSITUE 3346 02:30:11,480 --> 02:30:17,720 [INDISCERNIBLE] BY UJM, WHICH 3347 02:30:17,720 --> 02:30:19,040 GENERATE A 4 SIDE 3348 02:30:19,040 --> 02:30:22,320 [INDISCERNIBLE] OF THE 3349 02:30:22,320 --> 02:30:23,240 [INDISCERNIBLE] MUTATION, 3350 02:30:23,240 --> 02:30:25,480 INTRODUCING AS A HANDLE ON THE 3351 02:30:25,480 --> 02:30:26,840 TERMINAL SUGAR. 3352 02:30:26,840 --> 02:30:29,680 SO PRODUCT CAN BE READY TO 3353 02:30:29,680 --> 02:30:35,000 PURIFY, IN THE SOLUMKC. 3354 02:30:35,000 --> 02:30:40,040 AND HE ALSO FUNDED THE GTAC, 3355 02:30:40,040 --> 02:30:42,840 THIS CAN TAKE THE MODIFIED SUGAR 3356 02:30:42,840 --> 02:30:46,440 LIPID AS A DONOR AND YOU 3357 02:30:46,440 --> 02:30:53,720 COMPUEITATE, INCORNERATE IT'S 3358 02:30:53,720 --> 02:30:54,760 MODIFIED RABBINOSE WHICH WOULD 3359 02:30:54,760 --> 02:31:00,720 ALLOW THE FROZEN [INDISCERNIBLE] 3360 02:31:00,720 --> 02:31:03,320 BY THE CHEMISTRY. 3361 02:31:03,320 --> 02:31:05,120 THE 3 DIFFERENT SUGAR 3362 02:31:05,120 --> 02:31:09,760 [INDISCERNIBLE] WHICH IS THEOT 3363 02:31:09,760 --> 02:31:11,240 CARBON STRAIGHT, CARBON 5 AND 3364 02:31:11,240 --> 02:31:14,440 FOUND OUT THE DIFFERENT PROBE 3365 02:31:14,440 --> 02:31:17,200 CAN BE PERFERED BY THE DIFFERENT 3366 02:31:17,200 --> 02:31:19,160 PATIENTS FOR EXAMPLE, 3367 02:31:19,160 --> 02:31:25,880 [INDISCERNIBLE], OF THE PA, AND 3368 02:31:25,880 --> 02:31:27,240 [INDISCERNIBLE] SO THE PROBES, 3369 02:31:27,240 --> 02:31:29,160 HOW MANY ARE USED FOR 3370 02:31:29,160 --> 02:31:32,240 PUBLICATIONS FOR THIS CAN BE 3371 02:31:32,240 --> 02:31:34,680 USED FOR THE HYGIENES OF THE 3372 02:31:34,680 --> 02:31:40,640 DPLI CAN IN THE SEMESTER WALL, 3373 02:31:40,640 --> 02:31:42,560 MORE COMPLEX SYSTEM, FIRST THE 3374 02:31:42,560 --> 02:31:45,320 [INDISCERNIBLE], YOU INCORPORATE 3375 02:31:45,320 --> 02:31:46,280 THE 2 [INDISCERNIBLE] RABBINOSE 3376 02:31:46,280 --> 02:31:50,800 INTO THE TPHREU RACING KAN--KANA 3377 02:31:50,800 --> 02:31:52,040 OR THE [INDISCERNIBLE]. 3378 02:31:52,040 --> 02:31:54,400 WHEN TD TARGET GOES TO THE 3379 02:31:54,400 --> 02:31:56,600 MACROPHAGE CAN BE OBSERVED, YOU 3380 02:31:56,600 --> 02:31:59,240 CAN THAT FOR THE PROBE TO 3381 02:31:59,240 --> 02:32:07,400 MONITOR THE CHANGING, DURING THE 3382 02:32:07,400 --> 02:32:08,440 [INDISCERNIBLE]. 3383 02:32:08,440 --> 02:32:10,200 THIS GROUP TARGETED AND 3384 02:32:10,200 --> 02:32:14,880 [INDISCERNIBLE] 71 DEPENDENT 3385 02:32:14,880 --> 02:32:22,240 TARGET FOR EXAMPLE, THIS IS PF2, 3386 02:32:22,240 --> 02:32:25,080 AND SIDE WHAT 3 DEFINITE 3387 02:32:25,080 --> 02:32:27,280 POLYSACCHARIDE, SO IT'S A P A S 3388 02:32:27,280 --> 02:32:30,880 2, IS THE MOST ABUNDANT 1 WHICH 3389 02:32:30,880 --> 02:32:37,600 HAS THE HETEROCANNED WHAT. 3390 02:32:37,600 --> 02:32:40,360 THE HOW ALL THE [INDISCERNIBLE] 3391 02:32:40,360 --> 02:32:42,440 TO TRANSFER ACE, THAT STAND FROM 3392 02:32:42,440 --> 02:32:45,560 THE CHEMICAL INSIDE THE 3393 02:32:45,560 --> 02:32:48,760 MONOCELLULAR REPAID, IT PRODUCE 3394 02:32:48,760 --> 02:32:54,480 THIS NEOINATE ON THE SCALE. 3395 02:32:54,480 --> 02:32:55,800 THE GROUP ALSO CO INSIDE A 3396 02:32:55,800 --> 02:33:00,920 NUMBER OF THE [INDISCERNIBLE]. 3397 02:33:00,920 --> 02:33:05,040 THE [INDISCERNIBLE] ORGAN SAY 3398 02:33:05,040 --> 02:33:06,200 WHY, [INDISCERNIBLE] NEOINATE. 3399 02:33:06,200 --> 02:33:12,440 WE KNOW THIS IS A POLYMERASE 3400 02:33:12,440 --> 02:33:15,840 [INDISCERNIBLE], IS HARD TO DO 3401 02:33:15,840 --> 02:33:16,080 THE SKILL. 3402 02:33:16,080 --> 02:33:18,200 THIS GROUP IS VERY DEFICIENT IN 3403 02:33:18,200 --> 02:33:20,640 THE CHEMICAL APPROACH TO COUPLE 3404 02:33:20,640 --> 02:33:23,440 THE NEOINATE FORM THE 3405 02:33:23,440 --> 02:33:27,240 [INDISCERNIBLE] FOR OUR 3406 02:33:27,240 --> 02:33:27,520 ENGAGEMENT. 3407 02:33:27,520 --> 02:33:28,240 THE [INDISCERNIBLE] OR THE 1 3408 02:33:28,240 --> 02:33:32,760 PAGE THAT WE HAVE THE 3409 02:33:32,760 --> 02:33:33,240 HYDROLYSIS. 3410 02:33:33,240 --> 02:33:37,080 SO THIS APPROACHED BY THE 3411 02:33:37,080 --> 02:33:39,080 HYDROXIDE, SUGAR PLAT, THIS IS 3412 02:33:39,080 --> 02:33:42,560 ANOTHER SUGAR CONTAINED THE 3413 02:33:42,560 --> 02:33:42,760 GROUP. 3414 02:33:42,760 --> 02:33:44,520 THE REST OF THE WATER CAN BE 3415 02:33:44,520 --> 02:33:50,160 FINISHING IN HALF AN HOUR, AND 3416 02:33:50,160 --> 02:33:53,720 THE GROUP HAS IS A LAB 3417 02:33:53,720 --> 02:34:00,480 COLLABORATOR AND TEND TO SUFFER, 3418 02:34:00,480 --> 02:34:04,760 USE THIS APPROACH, 3419 02:34:04,760 --> 02:34:06,560 [INDISCERNIBLE] PROTEIN NEOINATE 3420 02:34:06,560 --> 02:34:09,320 WHICH IS A START FROM 3421 02:34:09,320 --> 02:34:11,760 THE--COUPLE THE TRI SACCHARIDE 3422 02:34:11,760 --> 02:34:17,440 WITH THE FLOW RATE. 3423 02:34:17,440 --> 02:34:18,040 [INDISCERNIBLE] DROKSIDE WITH 3424 02:34:18,040 --> 02:34:24,080 THE OTHER GROUP USING MUTATIONAL 3425 02:34:24,080 --> 02:34:29,520 2 ENZYME WITH 2 SUGARS, FOR THIS 3426 02:34:29,520 --> 02:34:31,480 NEOINATE, CURRENTLY IN THIS 3427 02:34:31,480 --> 02:34:32,960 GROUP IS WORKING OUT, YOU USE 3428 02:34:32,960 --> 02:34:35,240 THE SAME STRATEGIC PLAN EMGY TO 3429 02:34:35,240 --> 02:34:42,480 REACHING MUCH LONGER REPEATING 3430 02:34:42,480 --> 02:34:43,400 NEOINATE, AFTER 20. 3431 02:34:43,400 --> 02:34:44,480 OKAY, SO CURRENTLY WE ARE 3432 02:34:44,480 --> 02:34:47,720 WORKING ON THE PROTOCOL, AND WE 3433 02:34:47,720 --> 02:34:51,640 ARE DEVELOPMENT OUT IN THIS--FOR 3434 02:34:51,640 --> 02:34:53,920 THIS WEBSITE, AND FOR OTHER 3435 02:34:53,920 --> 02:34:58,960 PROTOCOL READY ANDY ARE ALSO 3436 02:34:58,960 --> 02:34:59,520 PRODUCING [INDISCERNIBLE]. 3437 02:34:59,520 --> 02:35:02,640 WHILE WORKING WITH PROFESSOR 3438 02:35:02,640 --> 02:35:05,720 [INDISCERNIBLE] AT UCD, AND TO 3439 02:35:05,720 --> 02:35:09,240 PRODUCE THE BIOMATERIALS, SINCE 3440 02:35:09,240 --> 02:35:11,800 THAT MATERIAL DPLI KAN--KANAS 3441 02:35:11,800 --> 02:35:12,480 AND OUR [INDISCERNIBLE]. 3442 02:35:12,480 --> 02:35:17,240 IS DOING THE CROSS VALIDATION, 3443 02:35:17,240 --> 02:35:23,600 ALSO CROSS VALIDATION, AND ALSO 3444 02:35:23,600 --> 02:35:25,960 TO PROTECT OKAY, FINALLY, I WILL 3445 02:35:25,960 --> 02:35:27,960 THANK ALL OF OUR TEAM MEMBERS, 3446 02:35:27,960 --> 02:35:30,640 ON THE THANK YOU FOR THIS WORK. 3447 02:35:30,640 --> 02:35:43,240 THANK YOU FOR YOUR PATIENCE. 3448 02:35:43,240 --> 02:35:43,800 [ APPLAUSE ] 3449 02:35:43,800 --> 02:35:47,520 >> THE NEXT SPEAKER WILL BE 3450 02:35:47,520 --> 02:35:59,960 ANDREW LEE FROM IMCS. 3451 02:35:59,960 --> 02:36:01,840 >> TESTING OKAY, THANK YOU, I AM 3452 02:36:01,840 --> 02:36:03,680 ANDREW LEE AND I THANK THE 3453 02:36:03,680 --> 02:36:05,120 ORGANIZER AND PAM FOR INVITING 3454 02:36:05,120 --> 02:36:06,800 ME HERE, IT'S MY FIRST TIME 3455 02:36:06,800 --> 02:36:08,240 PRESENTING IN FRONT OF THIS 3456 02:36:08,240 --> 02:36:12,400 AUDIENCE, I WILL CONFESS, I HAVE 3457 02:36:12,400 --> 02:36:13,400 NO GLYCO EXPERIENCE WHATSOEVER 3458 02:36:13,400 --> 02:36:15,240 AND WHAT AM I DOING HERE BUT 3459 02:36:15,240 --> 02:36:21,480 WE'LL ACTUALLY START WITH 1 3460 02:36:21,480 --> 02:36:23,000 SLIDE BACK WE ARE COLLABORATING 3461 02:36:23,000 --> 02:36:24,480 AND WE DID A FANTASTIC 3462 02:36:24,480 --> 02:36:26,840 INTRODUCTION, WE ARE PART OF 3463 02:36:26,840 --> 02:36:29,120 THAT ENZYME SCALE UP COMMERCIAL 3464 02:36:29,120 --> 02:36:29,360 ENTITY. 3465 02:36:29,360 --> 02:36:32,880 SO THE TITLE SAYS, TRANSITIONING 3466 02:36:32,880 --> 02:36:35,240 IN THE BENCH SCALE BACK TO 3467 02:36:35,240 --> 02:36:36,040 INDUSTRY SCALE FROM YOUR HANDS 3468 02:36:36,040 --> 02:36:37,800 BACK TO THE PENCH. 3469 02:36:37,800 --> 02:36:40,040 WHAT IMCS DOES IS WE DESIGN, 3470 02:36:40,040 --> 02:36:40,760 DEVELOP AND MANUFACTURE AND 3471 02:36:40,760 --> 02:36:41,040 DISTRIBUTE. 3472 02:36:41,040 --> 02:36:43,320 SO FROM THE START TO FINISH, OF 3473 02:36:43,320 --> 02:36:46,120 ALL ENZYME PRODUCTS AS WELL AS 3474 02:36:46,120 --> 02:36:46,680 THE CHROMATOGRAPHY PRODUCT. 3475 02:36:46,680 --> 02:36:50,320 AND IF YOU HAPPEN TO CONFUSE THE 3476 02:36:50,320 --> 02:36:54,640 ACRONYM, IMCS, THINK OF IT AS AI 3477 02:36:54,640 --> 02:36:57,440 AM A CUTE SCIENTIST, JUST 3478 02:36:57,440 --> 02:36:59,360 REMEMBER TO THE ACRONYM. 3479 02:36:59,360 --> 02:37:01,560 SO IMCS IS A PRETTY YOUNG 3480 02:37:01,560 --> 02:37:02,160 COMPANY. 3481 02:37:02,160 --> 02:37:06,440 WE STARTED IN 2013 AND WE 3482 02:37:06,440 --> 02:37:07,440 ACTUALLY FOCUSED ON 3483 02:37:07,440 --> 02:37:08,040 MICROCHROMATOGRAPHY PRODUCTS, 3484 02:37:08,040 --> 02:37:09,320 BUT WITHIN 6 MONTHS WE KICKED 3485 02:37:09,320 --> 02:37:12,440 OFF AND FOUND A BETA GLUE 3486 02:37:12,440 --> 02:37:15,040 CHRONODACE THAT TOOK OVER THE 3487 02:37:15,040 --> 02:37:16,480 URINE DRUG TESTING INDUSTRY. 3488 02:37:16,480 --> 02:37:17,760 SOPHISTICATEDY IN AMERICA IF YOU 3489 02:37:17,760 --> 02:37:19,160 KNOW PRESIDENT OPIOID CRISIS AND 3490 02:37:19,160 --> 02:37:23,040 YOU'RE IN DRUG TESTING IT IS 3491 02:37:23,040 --> 02:37:24,160 JUST AOF THED ROUGH ATOM 3492 02:37:24,160 --> 02:37:25,640 NATIONAL LIBRARY OF MEDICINEICAL 3493 02:37:25,640 --> 02:37:28,800 NUMBERS SO THIS ENZYME REPLACED 3494 02:37:28,800 --> 02:37:30,040 THE WHOLE URINE DRUG TESTING 3495 02:37:30,040 --> 02:37:32,800 MARKETS AND WE SCALE UP PRETTY 3496 02:37:32,800 --> 02:37:33,640 RAPIDLY SO WE'RE PRETTY FAMILIAR 3497 02:37:33,640 --> 02:37:36,040 WITH THE SAIL UP ENZYMES SO IN 3498 02:37:36,040 --> 02:37:42,000 TYPICAL ENZYME PRODO YOU 3499 02:37:42,000 --> 02:37:44,080 RECOGNIZE, WE WOULD DO AND 20 3500 02:37:44,080 --> 02:37:46,320 AND 21, THAT'S WHEN WE WERE 3501 02:37:46,320 --> 02:37:48,440 AWARDED THE NIH SBIR FAST TRACK 3502 02:37:48,440 --> 02:37:50,480 AWARD SO PHASE 1 AND PHASE 2 AND 3503 02:37:50,480 --> 02:37:53,080 THEN THE STTR AWARD WITH HIGH 3504 02:37:53,080 --> 02:38:00,840 YU, WE GOT IT IN 2021. 3505 02:38:00,840 --> 02:38:02,640 SO AS AN ENTREPRENEUR, THERE'S 3506 02:38:02,640 --> 02:38:03,960 NEW TECHNOLOGIES AND SEEING 3507 02:38:03,960 --> 02:38:05,680 EVERYBODY HERE, IT DOES BRING 3508 02:38:05,680 --> 02:38:06,720 SOME OF THIS MEMORY BACK BUT 3509 02:38:06,720 --> 02:38:12,360 THEN IN THE BACK OF MY MIND I'VE 3510 02:38:12,360 --> 02:38:14,080 ALSO EXPERIENCED SOME REALLY 3511 02:38:14,080 --> 02:38:16,040 SOME REALLY SCARY FOULS AND 3512 02:38:16,040 --> 02:38:18,800 LONG, LONG TOWN HILL TRAVELS, 3513 02:38:18,800 --> 02:38:20,160 TOO, BUT IT'S NOT ALL BAD 3514 02:38:20,160 --> 02:38:20,720 STORIES HERE. 3515 02:38:20,720 --> 02:38:22,840 I WILL POINT OUT EXCITING 3516 02:38:22,840 --> 02:38:23,840 RESULTS AND AND REALLY OUR 3517 02:38:23,840 --> 02:38:25,240 BACKGROUND WITH THE TIPS, WE 3518 02:38:25,240 --> 02:38:27,640 COLLABORATE WITH A LOT OF THESE 3519 02:38:27,640 --> 02:38:30,800 PHARMA COMPANIES. 3520 02:38:30,800 --> 02:38:33,680 PUBLISHED WITH PFIZER, SYNOFI 3521 02:38:33,680 --> 02:38:37,040 DID A WEBINAR FOR US AND ALSO WE 3522 02:38:37,040 --> 02:38:41,640 HAD A PUBLICATION FOR US. 3523 02:38:41,640 --> 02:38:44,520 AND THE [INDISCERNIBLE] THERE 3524 02:38:44,520 --> 02:38:49,120 ARE 700 LAPS TAKEN--THEY USE 3525 02:38:49,120 --> 02:38:50,440 THIS PRODUCT FOR DRUG AND THIS 3526 02:38:50,440 --> 02:39:06,840 IS A SERIES OF ENZYMES WE'RE 3527 02:39:06,840 --> 02:39:10,480 PLACING THIS TRANSFER ACES, THE 3528 02:39:10,480 --> 02:39:11,320 ACTIVATING GROUPS ARE THE 3529 02:39:11,320 --> 02:39:16,280 TRANSFER ACE AND THE 3530 02:39:16,280 --> 02:39:18,560 BIOSYNTHETIC ENZYMES. 3531 02:39:18,560 --> 02:39:19,840 YOU'VE ALREADY HASHED ABOUT 3532 02:39:19,840 --> 02:39:22,520 THOSE ENZYMES THAT ARE INVOLVED 3533 02:39:22,520 --> 02:39:25,440 IN THAT IN MOST OF THE ENZYME 3534 02:39:25,440 --> 02:39:28,600 WILL BE MANUFACTURED BY IMCs 3535 02:39:28,600 --> 02:39:30,040 AND COMMERCIALIZED SO FOR MORE 3536 02:39:30,040 --> 02:39:33,320 INFORMATION GO SEE THE POSTER 3537 02:39:33,320 --> 02:39:34,240 THAT DR. JOHN MACARTHUR WILL 3538 02:39:34,240 --> 02:39:37,520 PRESENT, HE DID HIS POST DOC 3539 02:39:37,520 --> 02:39:38,320 WITH C.CHEN AS WELL. 3540 02:39:38,320 --> 02:39:39,600 AND THE OTHER SET OF ENZYMES 3541 02:39:39,600 --> 02:39:41,440 THAT WILL BE COMMERCIALIZED ARE 3542 02:39:41,440 --> 02:39:43,520 ABOUT 10 DIFFERENT OTHER ENZYMES 3543 02:39:43,520 --> 02:39:48,000 THAT HELPS MAKE THE GLYCANS 3544 02:39:48,000 --> 02:39:48,200 HERE. 3545 02:39:48,200 --> 02:39:50,520 AND THE KIND OF THE COMPARISON 3546 02:39:50,520 --> 02:39:51,680 OF THE ENZYME SCALE UP THAT 3547 02:39:51,680 --> 02:39:53,560 WE'VE DONE IS THAT TYPICALLY IN 3548 02:39:53,560 --> 02:39:55,880 THE LAB SCALE, YOU DO YOUR FLASK 3549 02:39:55,880 --> 02:39:58,680 CULTURES AND WE'VE MIGRATED IT 3550 02:39:58,680 --> 02:40:02,040 TO A FED BATCH FERMENT AITIONZ 3551 02:40:02,040 --> 02:40:03,040 TECHNIQUES WITH OUR OWN MEDIA IN 3552 02:40:03,040 --> 02:40:09,280 THE CELLS AND WE TYPICALLY GET A 3553 02:40:09,280 --> 02:40:10,400 DESCENT 5 TO HUNDRED FOLD 3554 02:40:10,400 --> 02:40:11,840 IMPROVEMENT IN THE YEAR AND WE 3555 02:40:11,840 --> 02:40:14,360 WOULD TYPICALLY GO IN THE 3556 02:40:14,360 --> 02:40:17,840 QUAINTITYIS OF PRODUCTION FOR 3557 02:40:17,840 --> 02:40:19,040 THE LOW SCALE, AND AND THEN WE 3558 02:40:19,040 --> 02:40:20,840 HAVE TO MAKE SURE THEY ARE 3559 02:40:20,840 --> 02:40:22,360 PRODUCED IN THE FLASKS AS WELL 3560 02:40:22,360 --> 02:40:24,240 AS TRANSFERRED TO THE FED BATCH 3561 02:40:24,240 --> 02:40:26,640 SCALES ARE EQUIVALENT AND SO THE 3562 02:40:26,640 --> 02:40:28,480 PURITY OF BOTH THE FLASK 3563 02:40:28,480 --> 02:40:30,960 CULTURES AND THE FERMENTATION AS 3564 02:40:30,960 --> 02:40:32,640 SHOWN IN LANE 1 OR 2 FOR 1 OF 3565 02:40:32,640 --> 02:40:35,760 THE SILL O TRANSFER ACES, YOU 3566 02:40:35,760 --> 02:40:39,200 CAN SEE THE FERMENTATION METHOD 3567 02:40:39,200 --> 02:40:40,240 INTRODUCE EXTRA BANDS, THE 3568 02:40:40,240 --> 02:40:42,520 PURITY IS NOT AS PURE IN THE 3569 02:40:42,520 --> 02:40:44,080 SMALL SCALE BUT FOR BOTH SOLID 3570 02:40:44,080 --> 02:40:45,720 TRANSFER ACES OF THE SECOND 3571 02:40:45,720 --> 02:40:47,640 SOLID TRANSFER ACE OF LINK 2, 3 3572 02:40:47,640 --> 02:40:51,720 AND 4, THEY WILL HAVE COMPARABLE 3573 02:40:51,720 --> 02:40:53,480 LEVELS AND THE PERCENT 3574 02:40:53,480 --> 02:40:54,960 CONVERSION OF THE TRANSFER ACE 3575 02:40:54,960 --> 02:40:57,840 AND THE TRANSFERRING TO CMP 5 AC 3576 02:40:57,840 --> 02:41:00,320 IS EQUIV LEAPT WHETHER IT'S IN 3577 02:41:00,320 --> 02:41:02,520 PLAOF THESH R FLASK CULTURE, AS 3578 02:41:02,520 --> 02:41:05,040 YOU SEE IN THE TEAL AND THEN FOR 3579 02:41:05,040 --> 02:41:07,440 RED, THE SECOND TILE OF TRONS 3580 02:41:07,440 --> 02:41:08,760 FER ACE IS PRETTY EQUIVALENT 3581 02:41:08,760 --> 02:41:10,080 ACTIVITY WHETHER YOU CULTURE IT 3582 02:41:10,080 --> 02:41:14,040 IN THE FERMENTER OR IN THE 3583 02:41:14,040 --> 02:41:14,800 FLASK. 3584 02:41:14,800 --> 02:41:16,800 AND THEN THE--ONCE YOU GET THE 3585 02:41:16,800 --> 02:41:18,200 ENZYMES WE DO HAVE TO MAKE SURE 3586 02:41:18,200 --> 02:41:19,920 IT DOES GO OUT TO THE COMMUNITY. 3587 02:41:19,920 --> 02:41:21,640 AND WE HAVE TO MAKE SURE THAT IT 3588 02:41:21,640 --> 02:41:24,120 WORKS IN NOT ONLY GRADUATE 3589 02:41:24,120 --> 02:41:25,560 STUDENTS, I THINK JUSTIN POINTED 3590 02:41:25,560 --> 02:41:27,520 IT OUT THAT YOU MIGHT BE 3591 02:41:27,520 --> 02:41:28,800 SPENDING A HUNDRED DOLLARS ON AN 3592 02:41:28,800 --> 02:41:31,920 INN ZYME BUT IF THEY SPILL SOME 3593 02:41:31,920 --> 02:41:33,240 OR ACCIDENTALLY FREEZE SOME OR 3594 02:41:33,240 --> 02:41:34,640 ACTUALLY COOK IT, YOU DON'T WANT 3595 02:41:34,640 --> 02:41:36,440 IT TO BE INACTIVE AND YOU SPENT 3596 02:41:36,440 --> 02:41:38,520 THAT MONEY AND TIME PREPARING 3597 02:41:38,520 --> 02:41:39,160 FOR THAT REAGENT. 3598 02:41:39,160 --> 02:41:41,440 SO WE ACTUALLY GO PRETTY 3599 02:41:41,440 --> 02:41:42,840 EXTENSIVE ROUNDS OF FORMULATION 3600 02:41:42,840 --> 02:41:44,280 TO INSURE WE STABILIZE THAT 3601 02:41:44,280 --> 02:41:45,240 ENZYME. 3602 02:41:45,240 --> 02:41:49,760 SO HAVE YOU AN EXAMPLE HERE 3603 02:41:49,760 --> 02:41:51,200 HERE, DIFFERENT CONSULTS, 3604 02:41:51,200 --> 02:41:52,560 DIFFERENT BUFFERS AND TITRATIONS 3605 02:41:52,560 --> 02:41:54,640 AND WHAT'S PLOTTED IN THE Y-AXIS 3606 02:41:54,640 --> 02:41:56,880 HERE IS ELEVATED TEMPERATURE OR 3607 02:41:56,880 --> 02:42:00,400 MELT TEMPERATURES YOU HAVE A 3608 02:42:00,400 --> 02:42:01,600 MORE TABLE ENZYME. 3609 02:42:01,600 --> 02:42:02,960 THERE'S 1 INTERESTING POINT HERE 3610 02:42:02,960 --> 02:42:04,840 WITH THE ORANGE DOT ON THE LEFT 3611 02:42:04,840 --> 02:42:06,560 CORNER IS THAT THAT 1 PARTICULAR 3612 02:42:06,560 --> 02:42:09,480 SALT WITH THAT PH IS PRETTY 3613 02:42:09,480 --> 02:42:10,760 STABLE COMPARED TO EVERY OTHER 3614 02:42:10,760 --> 02:42:12,440 SALT WHERE IT TRENDS IN A 3615 02:42:12,440 --> 02:42:14,680 CONTINUOUS MANNER TOWARDS A MORE 3616 02:42:14,680 --> 02:42:15,240 BASIC PH, RIGHT? 3617 02:42:15,240 --> 02:42:16,920 BUT IF YOU COMPARE THAT 3618 02:42:16,920 --> 02:42:18,560 PARTICULAR SALT WITH OTHER 3619 02:42:18,560 --> 02:42:20,680 BUFFERS, EVEN AT THE HIGHER PH, 3620 02:42:20,680 --> 02:42:22,040 THAT SALT BUFFER DOES PROVIDE 3621 02:42:22,040 --> 02:42:24,840 MORE STABILITY FOR THAT ENZYME. 3622 02:42:24,840 --> 02:42:26,600 AND THEN AS IT GOES THROUGH 3623 02:42:26,600 --> 02:42:30,960 ROUNDS 1, 2, 3, 4, 5, YOU DO 3624 02:42:30,960 --> 02:42:32,720 SHIFT THAT MELTING AS SHOWN HERE 3625 02:42:32,720 --> 02:42:35,920 THIS IS A SINGLE WAVE LENGTH 3626 02:42:35,920 --> 02:42:37,640 LIGHT SCATTERING AT 256 WHERE IT 3627 02:42:37,640 --> 02:42:40,720 MEASURES NANO PARTICLES SO AS 3628 02:42:40,720 --> 02:42:42,840 THE PROTEIN UNFOLDS THE 3629 02:42:42,840 --> 02:42:44,080 PARTICULARRATES COME TOGETHER 3630 02:42:44,080 --> 02:42:47,160 AND IT WILL SCATTER LIGHT AT 266 3631 02:42:47,160 --> 02:42:49,960 AND YOU MAY ACROSS THE 3632 02:42:49,960 --> 02:42:51,200 TEMPERATURE AS THAT LIGHT 3633 02:42:51,200 --> 02:42:53,800 SCATTER INCREASES, YOU WANT TO 3634 02:42:53,800 --> 02:42:57,640 SEE THIS SHIFT MORE TO OUR 3635 02:42:57,640 --> 02:42:58,760 HIGHER TEMPERATURES, HENS MORE 3636 02:42:58,760 --> 02:43:00,720 STABLE PRODUCT, SO THIS IS YOUR 3637 02:43:00,720 --> 02:43:03,040 SURFACE PLATS YOU WILL GO WITH 3638 02:43:03,040 --> 02:43:05,720 YOUR PH AND YOUR SALT 3639 02:43:05,720 --> 02:43:06,840 CONCENTRATIONS, SO WITH THIS 3640 02:43:06,840 --> 02:43:09,000 CONSTRUCT A LOWER BUFFER SALT 3641 02:43:09,000 --> 02:43:11,520 WITH A HIGHER PH PROVIDED HIGHER 3642 02:43:11,520 --> 02:43:11,800 MELT TEMP. 3643 02:43:11,800 --> 02:43:14,320 SO A LOT OF THESE FORMULATIONS 3644 02:43:14,320 --> 02:43:16,480 WILL GO THROUGH FOR PRACTICALLY 3645 02:43:16,480 --> 02:43:17,680 21 DIFFERENT ENSWROIMS TO 3646 02:43:17,680 --> 02:43:19,760 SABLIZE IT AND THEN WE WOULD GO 3647 02:43:19,760 --> 02:43:21,160 THROUGH A LONG-TERM STABILITY 3648 02:43:21,160 --> 02:43:22,240 TEST, AT THERE DIFFERENT 3649 02:43:22,240 --> 02:43:23,160 TEMPERATURES AT MINIMUM SO HERE 3650 02:43:23,160 --> 02:43:26,640 IS 1 OF THE ENZYME CONSTRUCTS AT 3651 02:43:26,640 --> 02:43:28,200 NEGATIVE 20 DEGROOSS, 4-DEGREES 3652 02:43:28,200 --> 02:43:29,520 AND ROOM TEMPERATURE, 3653 02:43:29,520 --> 02:43:30,440 20-25-DEGREES, AND THE 3654 02:43:30,440 --> 02:43:33,320 ACTIVITIES ARE MEASURED BY 4 3655 02:43:33,320 --> 02:43:34,760 DIFFERENT ANALYSTS, CHEM KISTS, 3656 02:43:34,760 --> 02:43:41,080 ACROSS A 24 WEEK PERIOD. 3657 02:43:41,080 --> 02:43:45,160 SO ONCE YOU HAVE THE FORM AWGZ 3658 02:43:45,160 --> 02:43:47,040 WE HAVE A COMPONENT AND THE 3659 02:43:47,040 --> 02:43:48,520 ENZYME BETWEEN ENZYME A AND B AS 3660 02:43:48,520 --> 02:43:50,240 ALSO TO BE WORKED OUT SO THAT 3661 02:43:50,240 --> 02:43:52,560 HAVE YOU A ROBUST RANGE OF 3662 02:43:52,560 --> 02:43:53,640 PERFORMANCE SO HERE WHAT I'M 3663 02:43:53,640 --> 02:43:55,920 SHOW SUGGEST ABOUT A HUNDRED 3664 02:43:55,920 --> 02:43:57,320 MICROGAMS PER MILL CONCENTRATION 3665 02:43:57,320 --> 02:43:59,080 OF ENZYME A AND FURTHER DILUTED 3666 02:43:59,080 --> 02:44:02,240 10 FOLD AND THE RATIO OF ENZYME 3667 02:44:02,240 --> 02:44:03,960 B THAT'S TITRATED TO ENZYME A, 3668 02:44:03,960 --> 02:44:05,360 AND DEPEBDING ON THE TITRATION 3669 02:44:05,360 --> 02:44:08,520 YOU CAN HAVE A PRETTY EFFICIENT 3670 02:44:08,520 --> 02:44:10,040 CONVERSION OF THE ACTIVATED 3671 02:44:10,040 --> 02:44:13,240 SUGAR OR CAN YOU HAVE A PRETTY 3672 02:44:13,240 --> 02:44:17,720 LOW CONVERSION BUT YOU CAN SEE 3673 02:44:17,720 --> 02:44:18,600 THE DILUTION OF--LASER--NO. 3674 02:44:18,600 --> 02:44:22,560 THE ENZYME B IS NOT AS HIGHLY 3675 02:44:22,560 --> 02:44:22,840 REQUIRED. 3676 02:44:22,840 --> 02:44:26,400 A SMALL QUANTITY OF ENZYME B IS 3677 02:44:26,400 --> 02:44:26,640 THE PRICE. 3678 02:44:26,640 --> 02:44:30,360 MULTIPLIER IS 1 TO 1 RATIO OR 3679 02:44:30,360 --> 02:44:31,880 HUNDRED FOLD DILUTION, HUNDRED 3680 02:44:31,880 --> 02:44:34,080 FOLD DILUTION OF ENZYME BR IS 3681 02:44:34,080 --> 02:44:35,200 SUFFICIENT TO DRIVE THAT 3682 02:44:35,200 --> 02:44:36,840 REACTION AND ENZYME A, CAN YOU 3683 02:44:36,840 --> 02:44:38,360 ACTUALLY ACHIEVE YOUR REACTION 3684 02:44:38,360 --> 02:44:41,720 WITH 10 MICROGRAMS INSTEAD OF A 3685 02:44:41,720 --> 02:44:42,680 HUNDRED MICROGRAMS. 3686 02:44:42,680 --> 02:44:44,480 SO ULTIMATE GOAL MRILY TO 3687 02:44:44,480 --> 02:44:46,920 SUMMARIZE THE GRAND SCALE TO 3688 02:44:46,920 --> 02:44:48,920 KILOGRAM SCALE IS THE SWEET SPOT 3689 02:44:48,920 --> 02:44:52,200 FOR THE SYNTHETIC APPROACHES OR 3690 02:44:52,200 --> 02:44:53,680 ENZYMATIC SYNTHETIC APPROACHES 3691 02:44:53,680 --> 02:44:55,120 AND THEN WE'RE ENSURING THAT THE 3692 02:44:55,120 --> 02:44:56,520 ENZYMES ARE STABLE IN MULTIPLE 3693 02:44:56,520 --> 02:44:58,520 HANDS IT EVEN IN PROFESSOR'S 3694 02:44:58,520 --> 02:45:00,000 HANDS SO I KNOW GRAD STUDENTS 3695 02:45:00,000 --> 02:45:02,840 THEY DO A LOST WORK BUT, AND 3696 02:45:02,840 --> 02:45:04,680 SIMPLE TO EASY TO USE MOTE COLS, 3697 02:45:04,680 --> 02:45:10,000 SO THAT YOU GET ROBUSTY 3698 02:45:10,000 --> 02:45:12,120 REACTIONS, AND THE OPME GETS AN 3699 02:45:12,120 --> 02:45:13,200 AFFORDABLE REACTION SO THAT YOU 3700 02:45:13,200 --> 02:45:16,440 CAN MAKE HUNDREDS OF MILLIGRAMS 3701 02:45:16,440 --> 02:45:19,000 POTENTIALLY GRAMS OF THE 3702 02:45:19,000 --> 02:45:21,400 ASILATED GLYCANS AND AND THEN OF 3703 02:45:21,400 --> 02:45:23,240 COURSE PROFESSOR CHEN HOUNDS ME 3704 02:45:23,240 --> 02:45:25,800 ON THIS 1 AND I ADHERE ON IT 3705 02:45:25,800 --> 02:45:27,880 BECAUSE I MYSELF AM AN 3706 02:45:27,880 --> 02:45:31,080 ATTESTAMENT TO THIS, I AM A 3707 02:45:31,080 --> 02:45:33,840 NONSPECIALIST PROMISING TO MAKE 3708 02:45:33,840 --> 02:45:35,480 GANGLIO CITES AND OLIGO 3709 02:45:35,480 --> 02:45:35,800 SACCHARIDES. 3710 02:45:35,800 --> 02:45:37,720 SO I WOULD LIKE TO ACKNOWLEDGE 3711 02:45:37,720 --> 02:45:40,520 THE NIH PROGRAM, THE STTR, THE 3712 02:45:40,520 --> 02:45:42,840 R42 AND THE R44. 3713 02:45:42,840 --> 02:45:45,080 THAT'S THE SBIR AWARD AND THEN 3714 02:45:45,080 --> 02:45:46,720 THE IMCS TEAM THAT'S PICTURED 3715 02:45:46,720 --> 02:45:48,400 HERE THAT'S WORKING ON THE 3716 02:45:48,400 --> 02:45:49,960 VARIOUS ENZYMES HERE AND THEN 3717 02:45:49,960 --> 02:45:51,840 THE UCDAVIS, I KNOW THIS IS A 3718 02:45:51,840 --> 02:45:53,920 LOT SHORTER BUT I DID NOT GET 3719 02:45:53,920 --> 02:45:55,800 THE FULL LIST OF NAMES AND I 3720 02:45:55,800 --> 02:45:57,440 KNOW THERE'S PLENTY OF MORE 3721 02:45:57,440 --> 02:45:59,040 THAT'S ASSOCIATED WITH OUR UC 3722 02:45:59,040 --> 02:45:59,800 DAVIS TEAM. 3723 02:45:59,800 --> 02:46:07,280 SO THANK YOU SO MUCH 3724 02:46:07,280 --> 02:46:08,040 [ APPLAUSE ] 3725 02:46:08,040 --> 02:46:09,440 >> THANK YOU ANDREW FOR 3726 02:46:09,440 --> 02:46:11,440 REMINDING US THAT WE'RE ALL CUTE 3727 02:46:11,440 --> 02:46:12,640 IN SCIENCE IN 1 WAY OR ANOTHER. 3728 02:46:12,640 --> 02:46:17,800 SO THE NEXT TALK IS FROM RON 3729 02:46:17,800 --> 02:46:19,440 SCHNAAR, JOHNS HOPKINS SCHOOL OF 3730 02:46:19,440 --> 02:46:22,400 MEDICINE, HE WILL TELL US ABOUT 3731 02:46:22,400 --> 02:46:30,840 GANGLIO SIDE INTERACT OHM TOOL 3732 02:46:30,840 --> 02:46:31,040 KIT. 3733 02:46:31,040 --> 02:46:32,640 >> I HAVE SOME THANK YOUS, I 3734 02:46:32,640 --> 02:46:35,200 WANT TO THANK THE ENTIRE GROUP 3735 02:46:35,200 --> 02:46:38,680 UP HERE AND DOUG AND AMANDATORY 3736 02:46:38,680 --> 02:46:39,160 AND PAM. 3737 02:46:39,160 --> 02:46:42,240 MODEL CITIZEN HAVE BEEN THE FACE 3738 02:46:42,240 --> 02:46:43,040 OF THIS CONFERENCE. 3739 02:46:43,040 --> 02:46:46,200 I WANT TO THANK CHEN FOR 3740 02:46:46,200 --> 02:46:46,880 INTRODUCING GANGLIOICIDES SO I 3741 02:46:46,880 --> 02:46:49,080 DON'T HAVE TO DO AS MUCH OF 3742 02:46:49,080 --> 02:46:49,280 THAT. 3743 02:46:49,280 --> 02:46:52,440 AND FINELE, I HATE BEING THE 3744 02:46:52,440 --> 02:46:56,440 SPEAKER BEFORE LUNCH SO I WANT 3745 02:46:56,440 --> 02:46:59,080 TO THANK UMESH DESAI WHO'S AFTER 3746 02:46:59,080 --> 02:47:06,800 ME, WHOOPS, BACK. 3747 02:47:06,800 --> 02:47:09,440 >> THIS IS OUR TEAM. 3748 02:47:09,440 --> 02:47:12,440 THEY ARE HERE AND WILL BE 3749 02:47:12,440 --> 02:47:14,400 MANNING POSTER NUMBER 7 FOR 3750 02:47:14,400 --> 02:47:16,280 DETAILS OF WHERE WE'VE GONE WITH 3751 02:47:16,280 --> 02:47:19,160 THIS, SO THERE'S NO 3752 02:47:19,160 --> 02:47:20,320 GANGLIOICIDES, SO WE'RE 3753 02:47:20,320 --> 02:47:23,760 INTERESTED IN THEIR FUNCTIONS. 3754 02:47:23,760 --> 02:47:25,600 SO OF COURSE THIS IS JAM 3, 3755 02:47:25,600 --> 02:47:29,080 YOU'VE SEEN IT ALREADY, IT AS 3756 02:47:29,080 --> 02:47:30,400 THRI KAN--KANAS ATTACHED TO A 3757 02:47:30,400 --> 02:47:33,640 CERAMIDE AS YOU HEARD, 3758 02:47:33,640 --> 02:47:35,760 IMPORTANTLY, OF THE MANY 3759 02:47:35,760 --> 02:47:40,040 GANGLIOICIDES THAT ARE 3760 02:47:40,040 --> 02:47:44,560 AVAILABLE, IN NATURE THESE 8 ON 3761 02:47:44,560 --> 02:47:46,640 THE BOTTOM, I'M NOT GOING TO USE 3762 02:47:46,640 --> 02:47:49,520 THAT, I DON'T SEE THE OTHER 1 3763 02:47:49,520 --> 02:47:49,720 HERE. 3764 02:47:49,720 --> 02:47:51,960 THESE 8 DRIVE A LOT OF THE 3765 02:47:51,960 --> 02:48:03,360 FUNCTION THAT WE KNOW OF. 3766 02:48:03,360 --> 02:48:06,200 THESE 4G M1, DT1 A AND B, ARE 3767 02:48:06,200 --> 02:48:08,800 THE MAJOR GLYCANS OF THE BRAIN. 3768 02:48:08,800 --> 02:48:11,200 GM3 ASK A NONNEURONAL 3769 02:48:11,200 --> 02:48:12,280 GANGLIOICIDE INVOLVED IN 3770 02:48:12,280 --> 02:48:14,120 CONTROLLING METABOLISM AND AS AN 3771 02:48:14,120 --> 02:48:16,240 EXAMPLE GD2 IS A THERAPEUTIC 3772 02:48:16,240 --> 02:48:18,360 CANCER TARGET WITH AN ANTIBODY 3773 02:48:18,360 --> 02:48:18,920 IN THE MARKET. 3774 02:48:18,920 --> 02:48:23,760 SO THERE'S MANY PLACES WHERE 3775 02:48:23,760 --> 02:48:25,160 THESE HAVE FUNCTIONS, AND 3776 02:48:25,160 --> 02:48:26,560 IMPORTANT TO UNDERSTANDING THOSE 3777 02:48:26,560 --> 02:48:29,480 FUNCTIONS IS UNDERSTANDING WHERE 3778 02:48:29,480 --> 02:48:33,600 THEY ARE, THEY'RE EMBEDDED DEEP 3779 02:48:33,600 --> 02:48:36,640 IN THE PLESES MA MEMBRANES 3780 02:48:36,640 --> 02:48:40,880 PRIMARILY HEADING OUT INTO THE 3781 02:48:40,880 --> 02:48:41,400 EXTRA CELLULAR SPACE. 3782 02:48:41,400 --> 02:48:45,960 THEY CAN BE RELATIVELY LARGE, 3783 02:48:45,960 --> 02:48:49,920 50-ANGSTROMS, OUT FROM THE 3784 02:48:49,920 --> 02:48:51,760 MEMBRANE WHERE THEY CAN'T HAVE 3785 02:48:51,760 --> 02:48:52,600 MULTIPLE FUNCTIONS AND THOSE 3786 02:48:52,600 --> 02:48:57,720 FUNCTIONS WE THINK OF IT BEING 2 3787 02:48:57,720 --> 02:48:57,920 TYPES. 3788 02:48:57,920 --> 02:49:01,240 FIRST OF ALL THIS IS A STAINING 3789 02:49:01,240 --> 02:49:06,200 OF WHAT HAPPENS TO NERVE CELLS 3790 02:49:06,200 --> 02:49:08,720 IN CULTURE FOR GANGLIO SEEDS GU1 3791 02:49:08,720 --> 02:49:10,160 A AND B, AND YOU CAN SEE IT'S 3792 02:49:10,160 --> 02:49:11,800 ALL OVER THEIR SURFACES AND ALL 3793 02:49:11,800 --> 02:49:15,400 OVER ALL OF THEIR MEMBRANE 3794 02:49:15,400 --> 02:49:15,680 EXTENSIONS. 3795 02:49:15,680 --> 02:49:19,800 AND IN THE MEMBRANE, THE GANGLIO 3796 02:49:19,800 --> 02:49:23,560 SIDS CAN ACT LAT ROOLLY WITH 3797 02:49:23,560 --> 02:49:24,840 MOLECULES IN THEIR OWN AND ON 3798 02:49:24,840 --> 02:49:26,720 THE MEMBRANE ON THE LEFT TO 3799 02:49:26,720 --> 02:49:29,840 MODIFY THE ACTIVITY OF THOSE 3800 02:49:29,840 --> 02:49:31,400 PROTEINS. 3801 02:49:31,400 --> 02:49:34,760 AND THIS HAPPENS FOR INSTANCE 3802 02:49:34,760 --> 02:49:37,520 FOR RECEPTOR TYROSEEN CAINS,IS 3803 02:49:37,520 --> 02:49:40,000 CONTROL THE INSULIN RECEPTOR OF 3804 02:49:40,000 --> 02:49:40,960 GROWTH REPRESENT SEPTORSOR TRACK 3805 02:49:40,960 --> 02:49:43,280 A AND FOR PERHAPS 3806 02:49:43,280 --> 02:49:44,760 NEUROTRANSMITTER RECEPTORS. 3807 02:49:44,760 --> 02:49:48,440 IN ADDITION, THEY CAN ACT AS 3808 02:49:48,440 --> 02:49:49,360 RECOGNITION COLKIEWLS FOR 3809 02:49:49,360 --> 02:49:56,360 PROTEINS THAT ARE IN THE EXTRA 3810 02:49:56,360 --> 02:50:03,440 CELLULAR SPACE OR OTHER WITH THE 3811 02:50:03,440 --> 02:50:06,600 AXON AND BACTERIAL TOXINS ARE 3812 02:50:06,600 --> 02:50:09,000 SOLUBLE PROTEINS THAT BIND TO 3813 02:50:09,000 --> 02:50:10,440 GANGLIOICIDES, SO THIS IS WHAT 3814 02:50:10,440 --> 02:50:15,120 OUR GOAL IS ABOUT, IT'S ABOUT 3815 02:50:15,120 --> 02:50:17,680 UNDERSTANDINGLET IRPT ACTION OF 3816 02:50:17,680 --> 02:50:18,800 GANGLIOICIDES WITH PROTEINS TO 3817 02:50:18,800 --> 02:50:22,440 DRIVE ACTIVITY, AND WE WANT TO 3818 02:50:22,440 --> 02:50:23,160 [INDISCERNIBLE] GANGLIOICIDE 3819 02:50:23,160 --> 02:50:25,240 INTERACT OHM, TO BE A BROADLY 3820 02:50:25,240 --> 02:50:28,240 APPLICABLE SET OF PHOTO AFFINITY 3821 02:50:28,240 --> 02:50:31,800 CLICK GANGLIOICIDES TO ADENTIFY 3822 02:50:31,800 --> 02:50:35,520 GANGLIO SIDE INTERACTING TEENS 3823 02:50:35,520 --> 02:50:37,200 IN VARIOUS, BIOLODGE 8 HOURSICAL 3824 02:50:37,200 --> 02:50:38,240 TON TEXTS. 3825 02:50:38,240 --> 02:51:15,040 THE TEAM HAVE SUCCESSED IN THESE 3826 02:51:15,040 --> 02:51:18,960 3 GOALS IF WE PUT THE TAG ON THE 3827 02:51:18,960 --> 02:51:20,880 FATTY ACID, PART OF THE SERIES 3828 02:51:20,880 --> 02:51:23,240 POINTS MID IN THE MEMBRANE. 3829 02:51:23,240 --> 02:51:25,840 AND THAT WE HAVE SHORT TAGS, 3830 02:51:25,840 --> 02:51:28,800 THAT HAVE A [INDISCERNIBLE] AND 3831 02:51:28,800 --> 02:51:31,560 AN L-CINE FOR PHOTO AFFINITY 3832 02:51:31,560 --> 02:51:32,160 LABELING AND CLICK CHEMISTRY, 3833 02:51:32,160 --> 02:51:36,920 THIS CAN BE ON THE GATE KEEPER 3834 02:51:36,920 --> 02:51:41,240 LACTOSE AS SHOWN, THE ACID, AND 3835 02:51:41,240 --> 02:51:47,920 JAM 1 TAG B, THERE'S THE LONG 3836 02:51:47,920 --> 02:51:55,040 CHAIN REPLACEMENT FOR THE FATTY 3837 02:51:55,040 --> 02:51:55,640 ACID. 3838 02:51:55,640 --> 02:51:57,040 YELL AT ME TIMEWISE, OKAY? 3839 02:51:57,040 --> 02:51:58,320 THIS IS JUST EXAMPLES, I WILL 3840 02:51:58,320 --> 02:52:00,440 NOT SHOW YOU ALL THE THINGS 3841 02:52:00,440 --> 02:52:02,720 WE'VE MADE BUT FOR EXAMPLE, JAM 3842 02:52:02,720 --> 02:52:05,480 1 UPPER READ ARROW ON THE LEFT 3843 02:52:05,480 --> 02:52:08,240 WE CAN TARGET THE SIDE CHAIN 3844 02:52:08,240 --> 02:52:09,840 WITH [INDISCERNIBLE]. 3845 02:52:09,840 --> 02:52:11,520 TO GENERATE A UNIQUE ALDEHYDE 3846 02:52:11,520 --> 02:52:15,480 WHICH WE CAN CONVERT GOING DOWN 3847 02:52:15,480 --> 02:52:18,600 TO THE NEXT LINE TO THAT PRIMARY 3848 02:52:18,600 --> 02:52:22,640 AMINE AND THEN YOU JUST HAVE 3849 02:52:22,640 --> 02:52:28,880 PRIMARY AMINE TO INSTALL THE 3850 02:52:28,880 --> 02:52:29,120 L-KINE. 3851 02:52:29,120 --> 02:52:30,320 LIKEWISE, WEATHER ON THE TOP 3852 02:52:30,320 --> 02:52:33,360 LEFT, WE CAN TARGET THE GAL 3853 02:52:33,360 --> 02:52:35,920 ACTOSE 6 POSITION WITH THE GAL 3854 02:52:35,920 --> 02:52:38,560 ACTOSEOX DACE TO AGAIN MAKE A 3855 02:52:38,560 --> 02:52:39,400 UNIQUE ALDA HIDESSA HIDE, 3856 02:52:39,400 --> 02:52:48,240 CONVERTED TO AN AMINE AND 3857 02:52:48,240 --> 02:52:49,040 INSTALL OUR SHORT 3858 02:52:49,040 --> 02:52:49,600 [INDISCERNIBLE] THERE. 3859 02:52:49,600 --> 02:52:52,640 AND FINALLY, IN TERMS OF GENERAL 3860 02:52:52,640 --> 02:52:57,440 PROCEDURES, WE WERE LUCKY ENOUGH 3861 02:52:57,440 --> 02:53:00,000 TO GET AN EXPRESSING PLAZ MIDS 3862 02:53:00,000 --> 02:53:02,160 MIDOF SINGLE LIPID SERIES POINTS 3863 02:53:02,160 --> 02:53:05,760 MID MDA SOLACE WHICH CLIPS THE 3864 02:53:05,760 --> 02:53:07,680 FATTY ACID OFF, LEAVING US 3865 02:53:07,680 --> 02:53:10,040 PRIMARY AMINE AND 2 STEPS WE CAN 3866 02:53:10,040 --> 02:53:11,880 GO FROM THE LYSOGANG LIAISONEE 3867 02:53:11,880 --> 02:53:13,200 SID TO OUR IN TAG B. 3868 02:53:13,200 --> 02:53:19,280 ALL OF THESE HAVE BEEN PRODUCED 3869 02:53:19,280 --> 02:53:20,960 AND FINALLY WE CAN USE THE 3870 02:53:20,960 --> 02:53:22,440 SYNTHESIS, YOU HEARD SO MUCH 3871 02:53:22,440 --> 02:53:25,480 ABOUT TO MAKE MORE COMPLEX SCAN 3872 02:53:25,480 --> 02:53:29,520 OUT OF SIMPLER 1S. 3873 02:53:29,520 --> 02:53:34,560 SAMPLE HERE, GEM 1, TAG A, WE 3874 02:53:34,560 --> 02:53:35,320 TREAT WITH [INDISCERNIBLE], 3875 02:53:35,320 --> 02:53:37,480 TRANSFER ACE IN THE PRESENCE OF 3876 02:53:37,480 --> 02:53:40,560 SIALOGLYCANSIC ACID TO MAKE GD1 3877 02:53:40,560 --> 02:53:41,680 A [INDISCERNIBLE] TAG A. 3878 02:53:41,680 --> 02:53:42,960 AND OF COURSE WE DO THAT FOR TAG 3879 02:53:42,960 --> 02:53:45,640 B SO WE CAN EXPAND OUTWARDS IN 3880 02:53:45,640 --> 02:53:49,560 THIS WAY, USING CHEMO AND 3881 02:53:49,560 --> 02:53:51,000 SEMANTICS SYNTHESIS. 3882 02:53:51,000 --> 02:53:53,120 THESE ARE CHARACTERIZED BY THIN 3883 02:53:53,120 --> 02:53:54,160 LAYER CHROMATOGRAPHY ON THE 3884 02:53:54,160 --> 02:53:54,640 LEFT. 3885 02:53:54,640 --> 02:53:57,720 WE HAVE A NICE PURIFICATION 3886 02:53:57,720 --> 02:54:01,720 PROCEDURE THAT MITCHELL AND 3887 02:54:01,720 --> 02:54:04,000 GALLON HAVE OPTIMIZED PROTOON 3888 02:54:04,000 --> 02:54:06,040 AND CARBON NMR AND MASS 3889 02:54:06,040 --> 02:54:10,920 SPECTROMETRY, SO THESE ARE 3890 02:54:10,920 --> 02:54:11,360 CHARACTERIZED TAGS. 3891 02:54:11,360 --> 02:54:13,400 TO DELIVER THE CELL IS A TRICK, 3892 02:54:13,400 --> 02:54:16,400 BECAUSE IF YOU TAKE GANGLIO SIDS 3893 02:54:16,400 --> 02:54:18,840 THEY MAKE NICE CELLS AND IF YOU 3894 02:54:18,840 --> 02:54:20,840 ADD THEM, THEY GET TAKEN UP AND 3895 02:54:20,840 --> 02:54:22,720 SENT TO THE LYSOSOME, BUT WE HAD 3896 02:54:22,720 --> 02:54:25,480 FOUND IN OTHER STUDIES, THAT 3897 02:54:25,480 --> 02:54:29,240 DELIVERING THEM IN COMPLEX WITH 3898 02:54:29,240 --> 02:54:29,800 METHYL BATTA DEXTRAN SULFATE 3899 02:54:29,800 --> 02:54:31,760 TRON ON THE LEFT, DOES ENHANCE 3900 02:54:31,760 --> 02:54:34,800 THAT AND ENHANCES IT REALLY 3901 02:54:34,800 --> 02:54:35,000 WELL. 3902 02:54:35,000 --> 02:54:39,240 SO WE PREMIX THE METHYL BAIT 3903 02:54:39,240 --> 02:54:40,280 BATTA WITH THE PROBE, INCUBATE 3904 02:54:40,280 --> 02:54:41,760 IT WITH CELLS WASHED AND TO SEE 3905 02:54:41,760 --> 02:54:47,240 WHERE IT IS, WE CLICK THE ALKINE 3906 02:54:47,240 --> 02:54:53,520 WITH ALEKSA FLOOR 3907 02:54:53,520 --> 02:54:53,880 [INDISCERNIBLE]. 3908 02:54:53,880 --> 02:55:00,320 AND THESE ARE THE BEAUTIFUL 3909 02:55:00,320 --> 02:55:02,040 IMAGES WITH THE CARCINOMA CELLS, 3910 02:55:02,040 --> 02:55:04,200 YOU CAN SEE IT LOOKS LIKEOOSE ON 3911 02:55:04,200 --> 02:55:06,040 THE SURFACE, ON THE RIGHT IS TAG 3912 02:55:06,040 --> 02:55:07,480 B DUG WAY DOWN INTO THE MEMBRANE 3913 02:55:07,480 --> 02:55:09,920 AT THE VERY END, IT DOES STAIN, 3914 02:55:09,920 --> 02:55:12,240 USING THIS PROCEDURE, BUT WE 3915 02:55:12,240 --> 02:55:17,320 KNOW IT'S THERE BECAUSE IT TAGS 3916 02:55:17,320 --> 02:55:17,680 PROTEINS. 3917 02:55:17,680 --> 02:55:19,000 GALLON AND MITCHELL WENT FURTHER 3918 02:55:19,000 --> 02:55:23,920 TO PROVE THIS IS ON THE SURFACE 3919 02:55:23,920 --> 02:55:26,600 BY TAKING GM 3 TAG A LOADED IN 3920 02:55:26,600 --> 02:55:29,760 THIS CASE, A431 SEMESTERS AND 3921 02:55:29,760 --> 02:55:31,440 TREATING THOSE WITH 3922 02:55:31,440 --> 02:55:32,080 SIALOGLYCANSITTICS WHICH DOESN'T 3923 02:55:32,080 --> 02:55:37,840 GET INSIDE THE CELLS AND THEY'RE 3924 02:55:37,840 --> 02:55:38,440 ALL ACCESSIBLE TO 3925 02:55:38,440 --> 02:55:41,440 [INDISCERNIBLE] AND THE TAG. 3926 02:55:41,440 --> 02:55:43,600 AND TO SHOW YOU THIS IS 3927 02:55:43,600 --> 02:55:46,400 TRANSFERAL BOTH TO OTHER CELL 3928 02:55:46,400 --> 02:55:47,640 TYPES, THIS IS NEUROBLASTOMA 3929 02:55:47,640 --> 02:55:49,800 CELLS, CAN YOU SEE THE DIFFERENT 3930 02:55:49,800 --> 02:55:51,520 MORPHOLOGY, BEAUTIFULLY STAINED 3931 02:55:51,520 --> 02:55:51,880 ON THE SURFACE. 3932 02:55:51,880 --> 02:55:56,240 STEP WOP SYNTHESIZE THEM, STEP 3933 02:55:56,240 --> 02:56:00,320 2, DELIVER THEM, STEP 3, TAG 3934 02:56:00,320 --> 02:56:02,960 PROTEINS SO WE'RE DOING, WE'RE 3935 02:56:02,960 --> 02:56:05,200 LOADING THE CELLS WITH OUR 3936 02:56:05,200 --> 02:56:08,520 METHYL CYCLE DEXTRIN GANG 3937 02:56:08,520 --> 02:56:10,240 BIOICIDE PROBE, WASHING THEM, 3938 02:56:10,240 --> 02:56:12,440 EXPOSING THEM TO UV TO ACTIVATE 3939 02:56:12,440 --> 02:56:17,000 THE PROTEINS, CLICK ON A BIOTIN 3940 02:56:17,000 --> 02:56:18,400 AICIDE SOLUBLIZE, ANALYZE AND 3941 02:56:18,400 --> 02:56:20,400 CAPTURE. 3942 02:56:20,400 --> 02:56:23,760 THIS IS AN EXAMPLE OF THE CELLS, 3943 02:56:23,760 --> 02:56:25,560 TREATED WITH THE 3 DIFFERENT 3944 02:56:25,560 --> 02:56:29,840 TAGS AT THE TOP FOR NO PROBE, WE 3945 02:56:29,840 --> 02:56:31,440 THEN SOLUBLIZE THE PROTEINS, RAN 3946 02:56:31,440 --> 02:56:34,320 THEM ON A GEL AND USED CHOLERA 3947 02:56:34,320 --> 02:56:37,240 TAXIN A WHICH RECOGNIZES JAM 1 3948 02:56:37,240 --> 02:56:38,840 AS THE PROBE AND CAN YOU SEE, WE 3949 02:56:38,840 --> 02:56:40,240 PICKED UP SOME NUMBER OF 3950 02:56:40,240 --> 02:56:45,040 PROTEINS IN THE HUNDRED TO 175 3951 02:56:45,040 --> 02:56:46,000 KILODALTON RANGE THAT 3952 02:56:46,000 --> 02:56:53,360 INTERACTING FROM THESE CELLS IF 3953 02:56:53,360 --> 02:56:55,760 WE--I SHOULD BACK AND SAY, YOU 3954 02:56:55,760 --> 02:57:22,200 SEE NO TRAINING WITH GM3 OR A 1 3955 02:57:22,200 --> 02:57:24,200 TAG A AND TAG B PICK UP 3956 02:57:24,200 --> 02:57:25,680 DIFFERENT PROTEINS FOR JAM 1 3957 02:57:25,680 --> 02:57:27,000 THOSE RIGHT DOWN AT THE BOTTOM. 3958 02:57:27,000 --> 02:57:29,400 WE BELIEVE WE HAVEN'T CONFIRMED 3959 02:57:29,400 --> 02:57:34,200 THIS, MAY BE SMALL TETRASPANINS, 3960 02:57:34,200 --> 02:57:36,400 THAT'S IN HOCKA MAURI OSHEROFFY 3961 02:57:36,400 --> 02:57:38,400 PROPOSED INTERACTING WITH 3962 02:57:38,400 --> 02:57:40,840 GAIPGLIOICIDES IN THE PLANE OF 3963 02:57:40,840 --> 02:57:47,400 THE IDEA TAG B AND TRICK THE 3964 02:57:47,400 --> 02:57:48,440 [INDISCERNIBLE] AND THEY ARE NOW 3965 02:57:48,440 --> 02:57:55,240 AT THE POINT OF CAPTURING FOR 3966 02:57:55,240 --> 02:57:55,720 THE [INDISCERNIBLE]. 3967 02:57:55,720 --> 02:57:58,040 WITH THAT I WILL FINISH, PLEASE 3968 02:57:58,040 --> 02:57:59,920 GO TO POSTER 7, THIS WORK HAS 3969 02:57:59,920 --> 02:58:02,600 REALLY BEEN DRIVEN BY 2 PEOPLE, 3970 02:58:02,600 --> 02:58:05,040 [INDISCERNIBLE] AND MITCHELL 3971 02:58:05,040 --> 02:58:07,080 PORTER, AND WE HAVE HAD 3972 02:58:07,080 --> 02:58:12,440 ESSENTIAL TOOLS PROVIDED TO US 3973 02:58:12,440 --> 02:58:13,960 BY DR. CHEN AS MENTIONED AND 3974 02:58:13,960 --> 02:58:16,800 ANDROGEN LIEU LEE WHO WE JUST 3975 02:58:16,800 --> 02:58:25,240 HEARD FROM AND MACARTHUR AT. 3976 02:58:25,240 --> 02:58:28,840 >> AND NOW FOR THE LAST SPEAKER 3977 02:58:28,840 --> 02:58:29,760 BEFORE LUNCH-- 3978 02:58:29,760 --> 02:58:51,760 >> WE HAVE 4 MORE MINUTES. 3979 02:58:51,760 --> 02:58:54,760 >> THE LAST SPEAKER IS UMESH 3980 02:58:54,760 --> 02:58:57,440 DESAI, WHO WILL TALK ABOUT TOOL 3981 02:58:57,440 --> 02:58:59,240 FOR PREDICTING GRIEK O 3982 02:58:59,240 --> 02:59:00,040 SOPHISTICATED MINE DPLI 3983 02:59:00,040 --> 02:59:01,360 KAN--KANA RECOGNITION OF 3984 02:59:01,360 --> 02:59:01,640 PROTEINS. 3985 02:59:01,640 --> 02:59:02,600 >> HELLO EVERYBODY, I WOULD LIKE 3986 02:59:02,600 --> 02:59:04,880 TO BEGIN BY THANKING PAM AND THE 3987 02:59:04,880 --> 02:59:06,840 CALL AND AMANDA AND EVERYBODY 3988 02:59:06,840 --> 02:59:09,080 CONNECTED WITH THE COMMON FUND 3989 02:59:09,080 --> 02:59:17,560 TO GIVE ME THIS OPPORTUNITY TO 3990 02:59:17,560 --> 02:59:19,560 ESSENTIALLY TELL YOU ABOUT WHAT 3991 02:59:19,560 --> 02:59:21,240 WE ACCOMPLISHED OVER THE LAST 3 3992 02:59:21,240 --> 02:59:22,440 YEARS AND ALSO IBDICATE TO WHERE 3993 02:59:22,440 --> 02:59:51,640 WE ARE HEADING SO, THE 3994 02:59:51,640 --> 02:59:54,040 SPECIFICALLY TRYING TO MOVE THE 3995 02:59:54,040 --> 02:59:56,680 IDEA THAT GLYCO GLYCANS CAN 3996 02:59:56,680 --> 02:59:58,240 ACTUALLY SERVE WELL AS DRUGS 3997 02:59:58,240 --> 03:00:01,960 BECAUSE THERE ARE VERY, VERY FEW 3998 03:00:01,960 --> 03:00:03,960 THAT ARE YOU KNOW HAVE SUCCEEDED 3999 03:00:03,960 --> 03:00:28,440 AS DRUGS AND I THINK WE THESE 4000 03:00:28,440 --> 03:00:29,640 WILL MAKE IT MORE FEASIBLE AND 4001 03:00:29,640 --> 03:00:30,480 THE SECOND PART OF THIS PROGRAM 4002 03:00:30,480 --> 03:00:32,160 WAS TO USE THIS TOOL AND 4003 03:00:32,160 --> 03:00:33,680 ESSENTIALLY HELP YOU KNOW 4004 03:00:33,680 --> 03:00:36,520 BIOLOGISTS WHO ARE NOT REALLY 4005 03:00:36,520 --> 03:00:38,160 TOO FAMILIAR WITH THE 4006 03:00:38,160 --> 03:00:45,920 COMPUTATIONAL ASPECTS TO COME UP 4007 03:00:45,920 --> 03:00:49,160 WITH IN BIOLOGY, SO THIS IS 4008 03:00:49,160 --> 03:00:52,440 ACTUALLY A 20 STEP TO BIG FOR 4009 03:00:52,440 --> 03:00:53,960 MOST PEOPLE BUT IF YOU REALLY 4010 03:00:53,960 --> 03:00:57,080 THINK FROM A BIOLOGIST 4011 03:00:57,080 --> 03:00:58,360 PERSPECTIVE THAT WHAT A 4012 03:00:58,360 --> 03:00:59,840 BIOLOGIST WANT TO KNOW HAVING 4013 03:00:59,840 --> 03:01:01,440 NOT SO MUCH UNDERSTANDING ABOUT 4014 03:01:01,440 --> 03:01:04,840 YOU KNOW GAGS AND THINGS OF THAT 4015 03:01:04,840 --> 03:01:07,320 SORT, NUMBER 1, YOU KNOW DOES MY 4016 03:01:07,320 --> 03:01:09,680 PARTICULAR PROTEIN BIND TO GAGS, 4017 03:01:09,680 --> 03:01:11,560 THAT'S THE BASIC QUESTION, IF IT 4018 03:01:11,560 --> 03:01:13,640 DOES BIND TO GAG THEN WHERE DOES 4019 03:01:13,640 --> 03:01:15,240 IT BIND, IS THERE A SPECIFIC 4020 03:01:15,240 --> 03:01:24,640 SORT OF SIGHT OF BINDING THEN 4021 03:01:24,640 --> 03:01:26,560 WHAT GAG WOULD PREFERENTIALLY 4022 03:01:26,560 --> 03:01:29,440 BIND TO THAT PARTICULAR SITE? 4023 03:01:29,440 --> 03:01:31,000 AND ASSUMING THAT ALL OF THESE 4024 03:01:31,000 --> 03:01:32,160 ANSWERS COME THROUGH, TRYING TO 4025 03:01:32,160 --> 03:01:34,240 UNDERSTAND AS TO WHETHER INVIVO 4026 03:01:34,240 --> 03:01:36,640 IN REAL LIFE SITUATION WHERE 4027 03:01:36,640 --> 03:01:37,720 THAT COMPLEX REMAINS STABLE AND 4028 03:01:37,720 --> 03:01:40,240 IS LIKELY TO INDUCE THE FUNCTION 4029 03:01:40,240 --> 03:01:43,360 THAT WE THINK IS LIKELY TO 4030 03:01:43,360 --> 03:01:48,840 INDUCE, SO THE WE HAVE 4031 03:01:48,840 --> 03:01:53,080 DEVELOPED--WHOOPS LET'S GO BACK. 4032 03:01:53,080 --> 03:01:54,360 TRIES TO ADDRESS THE 3 QUESTIONS 4033 03:01:54,360 --> 03:01:56,680 THAT ARE HIGHLIGHTED HERE, DOES 4034 03:01:56,680 --> 03:01:59,520 MY PROTEIN BIND TO GAG, IS A GAG 4035 03:01:59,520 --> 03:02:01,600 SITE IS WHAT WOULD 4036 03:02:01,600 --> 03:02:03,600 PREFERENTIALLY RECOGNIZE THAT IS 4037 03:02:03,600 --> 03:02:05,120 IT'S PULT MULTIPLE STEPS BUT WE 4038 03:02:05,120 --> 03:02:06,440 HAVE TRIED TO AUTOMATE IT IN 4039 03:02:06,440 --> 03:02:08,480 SUCH A FASHION THAT WE CAN 4040 03:02:08,480 --> 03:02:10,600 AUTOMATE IT IN A LIBRARY OF 4041 03:02:10,600 --> 03:02:12,880 SEQUENCES SOMEWHERE AROUND A 4042 03:02:12,880 --> 03:02:13,840 THOUSAND THOUSAND DIFFERENT 4043 03:02:13,840 --> 03:02:14,080 SEQUENCES. 4044 03:02:14,080 --> 03:02:15,960 SO THIS IS ESSENTIALLY THE 4045 03:02:15,960 --> 03:02:19,280 BOTTOM LINE IN TERMS OF WHAT THE 4046 03:02:19,280 --> 03:02:21,040 ALGORITHM TRIES TO DO. 4047 03:02:21,040 --> 03:02:29,400 NUMBER 1 IT BUILDS A LIBRARY OF 4048 03:02:29,400 --> 03:02:42,400 SEQUENCES THAT YOU WANT--AND 4049 03:02:42,400 --> 03:02:43,640 THESE SEQUENCES ARE AVAILABLE 4050 03:02:43,640 --> 03:02:44,720 FROM [INDISCERNIBLE] AND DPLI 4051 03:02:44,720 --> 03:02:45,440 KAN--KANA THERAPEUTICS AND WE 4052 03:02:45,440 --> 03:02:47,800 HOPE WE CAN ADD ANYBODY WHO 4053 03:02:47,800 --> 03:02:51,120 MAKES THESE GAG SEQUENCES 4054 03:02:51,120 --> 03:02:51,840 COMMERCIALLY AVAILABLE, WE WOULD 4055 03:02:51,840 --> 03:02:56,040 ADD IN THE FUTURE. 4056 03:02:56,040 --> 03:02:57,840 WE WOULD SCREEN ASSUMING WE 4057 03:02:57,840 --> 03:02:59,040 DON'T KNOW ANYTHING ABOUT GAG 4058 03:02:59,040 --> 03:03:00,800 INTERACTING WITH THE PROTEIN, 4059 03:03:00,800 --> 03:03:01,960 ANYTHING ABOUT PROTEIN BINDING 4060 03:03:01,960 --> 03:03:04,240 SITES WE VALID TO SCREEN THE 4061 03:03:04,240 --> 03:03:05,800 ENTIRE PROTEIN SURFACE FOR THE 4062 03:03:05,800 --> 03:03:09,000 POSSIBILITY OF GAG BINDING SO 4063 03:03:09,000 --> 03:03:10,920 DO--MULTIPLE BINDING SIGHTS AND 4064 03:03:10,920 --> 03:03:13,240 THEN OBVIOUSLY DO A WHOLE LOT OF 4065 03:03:13,240 --> 03:03:14,560 COMPUTATIONAL WORK, WE HAVE 4066 03:03:14,560 --> 03:03:17,480 DEVELOPED A TECHNOLOGY OF USING 4067 03:03:17,480 --> 03:03:21,080 2 DIFFERENT FILTERS AND 4068 03:03:21,080 --> 03:03:22,520 TRADITIONALLY COMPUATIONAL WORK 4069 03:03:22,520 --> 03:03:24,840 UTILIZES 1 FILTERS, WE HAVE DONE 4070 03:03:24,840 --> 03:03:28,240 2 FILTERS BECAUSE OF THE NUANCES 4071 03:03:28,240 --> 03:04:04,920 ABOUT GAG PROTEIN INTERACTS 4072 03:04:04,920 --> 03:04:06,000 OF--WHICH MIGHT EXIST INTO 4073 03:04:06,000 --> 03:04:08,200 NATURE IN 2 BINS, 1 IS A BIN 4074 03:04:08,200 --> 03:04:10,240 WHERE YOU HAVE SPECIFIC 4075 03:04:10,240 --> 03:04:12,720 SEQUENCES, THE SECOND WOULD BE 4076 03:04:12,720 --> 03:04:13,520 NONSPECIFIC SEQUENCES, JUST 4077 03:04:13,520 --> 03:04:17,640 BECAUSE I AM A MEDICINAL 4078 03:04:17,640 --> 03:04:19,000 CHEMIST, I'M A DRUG DISCOVERY 4079 03:04:19,000 --> 03:04:20,200 PERSON, I'M MORE ABOUTED IN 4080 03:04:20,200 --> 03:04:22,040 THESE SEQUENCES BECAUSE THERE IS 4081 03:04:22,040 --> 03:04:24,040 A TENDENCY TO CONVERT THEM INTO 4082 03:04:24,040 --> 03:04:25,200 POSSIBILITIES PRETTY HIGH TO 4083 03:04:25,200 --> 03:04:26,920 CONSECRETER THEM INTO DRUGS. 4084 03:04:26,920 --> 03:04:27,760 UNDER ANY CIRCUMSTANCES THIS IS 4085 03:04:27,760 --> 03:04:35,200 THE TOOL THAT IS AVAILABLE FOR 4086 03:04:35,200 --> 03:05:09,680 ALL NONPROFIT/EDUCATIONAL 4087 03:05:09,680 --> 03:05:13,400 INSTITUTION--AND THIS OPERATION 4088 03:05:13,400 --> 03:05:14,040 OF THIS WEBSITE ACTUALLY 4089 03:05:14,040 --> 03:05:15,640 PRESENTED IN THE PAST SO I'M NOT 4090 03:05:15,640 --> 03:05:17,080 GOING TO GO INTO DETAIL BUT THE 4091 03:05:17,080 --> 03:05:19,040 BOTTOM LINE IS THAT THERE IS A 4092 03:05:19,040 --> 03:05:20,960 TUTORIAL THAT IS AVAILABLE, 4093 03:05:20,960 --> 03:05:22,440 THAT'S THE TUTORIAL, YOU LOG ON 4094 03:05:22,440 --> 03:05:24,160 TO THAT SITE, YOU CREATE A USER 4095 03:05:24,160 --> 03:05:26,240 NAME, YOU USE THE TUTORIAL, TO 4096 03:05:26,240 --> 03:05:28,320 ESSENTIALLY YOU KNOW SET UP THE 4097 03:05:28,320 --> 03:05:30,560 INDIVIDUAL RUNS AND IT'S 4098 03:05:30,560 --> 03:05:31,280 ESSENTIALLY FULLY AUTOMATED, 4099 03:05:31,280 --> 03:05:34,320 THERE ARE ONLY 5 STEPS, THE KEY 4100 03:05:34,320 --> 03:05:35,080 ASPECT IS REALLY INTERPRETATION 4101 03:05:35,080 --> 03:05:36,160 AND THAT'S WHERE YOU PROBABLY 4102 03:05:36,160 --> 03:05:37,680 NEED TO READ SOME OF THE PAPERS 4103 03:05:37,680 --> 03:05:38,840 THAT WE'RE PUBLISHED ON THE 4104 03:05:38,840 --> 03:05:42,280 TOPIC OF HOW TO REALLY 4105 03:05:42,280 --> 03:05:42,560 INTEGRATE. 4106 03:05:42,560 --> 03:05:48,000 BUT, AS OF NOW, I THINK WE PUT 4107 03:05:48,000 --> 03:05:49,840 THIS UP IN A YEAR, MAYBE 14 4108 03:05:49,840 --> 03:05:54,320 MONTHS AGO AND WE HAVE HAD 28 4109 03:05:54,320 --> 03:05:56,160 UNIQUE USERS, ABOUT 283 JOBS AS 4110 03:05:56,160 --> 03:05:58,160 OF LAST WEEK, YOU KNOW AND I 4111 03:05:58,160 --> 03:06:01,600 THINK I AM EXTREMELY HAPPY THAT 4112 03:06:01,600 --> 03:06:04,440 AN UNDERGRADUATE STUDENT WAS 4113 03:06:04,440 --> 03:06:06,280 ESSENTIALLY ABLE TO DO THE 4114 03:06:06,280 --> 03:06:08,160 RESEARCH USING THE WEB TOOL WITH 4115 03:06:08,160 --> 03:06:14,040 PRACTICALLY NO, YOU KNOW INPUT, 4116 03:06:14,040 --> 03:06:15,040 PRACTICALLY NO INPUT THAT 4117 03:06:15,040 --> 03:06:17,000 DOESN'T MEAN NO INPUT BUT 4118 03:06:17,000 --> 03:06:19,160 PRACTICALLY NO INPUT FROM 1 OF 4119 03:06:19,160 --> 03:06:19,920 MY LEADING SCIENTISTS. 4120 03:06:19,920 --> 03:06:20,800 FOURTEEN INSTITUTIONS IN THE 4121 03:06:20,800 --> 03:06:22,760 U.S. HAVE USED IT, THESE ARE 4122 03:06:22,760 --> 03:06:24,720 ACROSS THE ENTIRE U.S., FOR 4123 03:06:24,720 --> 03:06:28,160 VARIOUS PURPOSES WE DON'T KNOW 4124 03:06:28,160 --> 03:06:29,160 EXACTLY WHAT EXPERIMENTS THEY 4125 03:06:29,160 --> 03:06:31,040 WERE DOG BUT HOPE LOOFULLY THEY 4126 03:06:31,040 --> 03:06:32,120 GOT SOMETHING GOT OUT OF IT AND 4127 03:06:32,120 --> 03:06:34,880 WE WILL SEE THAT THIS PARTICULAR 4128 03:06:34,880 --> 03:06:37,160 WEB SERVER WILL BE RECOGNIZED IN 4129 03:06:37,160 --> 03:06:37,920 FUTURE PUBLICATIONS. 4130 03:06:37,920 --> 03:06:40,400 AS FAR AS MY GROUP AND 4131 03:06:40,400 --> 03:06:42,040 ESSENTIALLY TRYING TO HELP 4132 03:06:42,040 --> 03:06:44,920 BIOLOGISTS WE HAVE STUDIED 4133 03:06:44,920 --> 03:06:45,840 SEVERAL DIFFERENT PROTEINS, YOU 4134 03:06:45,840 --> 03:06:47,640 KNOW STUDIED A BUNCH OF 4135 03:06:47,640 --> 03:06:48,840 PROTEINS, 1 OF THE LAST, 2 4136 03:06:48,840 --> 03:06:54,240 YEARS, 1 AND HALF YEARS, WE HAVE 4137 03:06:54,240 --> 03:06:56,800 DONE YOU KNOW WORK ON THE LAST 4138 03:06:56,800 --> 03:06:58,640 FEW DAYS, TRANSFER ACE 1, CHEMO 4139 03:06:58,640 --> 03:07:00,840 KINES AND YOU HEARD TALK FROM 4140 03:07:00,840 --> 03:07:01,480 [INDISCERNIBLE] YESTERDAY ABOUT 4141 03:07:01,480 --> 03:07:03,720 SOME OF THE WORK THAT WE HAVE 4142 03:07:03,720 --> 03:07:06,720 DONE WITH HIM. 4143 03:07:06,720 --> 03:07:09,600 TGF BETA 2, SPIKE GLYCO PROTEIN 4144 03:07:09,600 --> 03:07:11,440 FOR SARS-COV-2, WE HAVE A PAPER 4145 03:07:11,440 --> 03:07:15,200 IN CURRENTLY IN REVIEW WITH 4146 03:07:15,200 --> 03:07:18,360 IGF 1 R, WITH IS A TYROSINE 4147 03:07:18,360 --> 03:07:21,400 KINASE, THAT BINDS TO IGF1 R AND 4148 03:07:21,400 --> 03:07:25,040 NRP 1 WHICH IS CURRENTLY IN 4149 03:07:25,040 --> 03:07:25,640 PROGRESS. 4150 03:07:25,640 --> 03:07:28,000 THESE ARE THE 4151 03:07:28,000 --> 03:07:30,600 BIOLOGISTS/BIOCHEMISTS THAT WE 4152 03:07:30,600 --> 03:07:38,440 HAVE TRIED TO HELP, SHE WORKS ON 4153 03:07:38,440 --> 03:07:39,840 CYSTIC FIBROSIS AND SOME OF YOU 4154 03:07:39,840 --> 03:07:45,160 MAY KNOW HIM WITH REGARD TO THE 4155 03:07:45,160 --> 03:07:47,440 TRANSFER ACE, KRISHNA, YOU HEARD 4156 03:07:47,440 --> 03:07:52,040 HIM, AND [INDISCERNIBLE] WORKS 4157 03:07:52,040 --> 03:07:54,440 ON SARS-COV-2, P A TEL IS 4158 03:07:54,440 --> 03:07:58,480 WORKING ON IGF-1 R AND 4159 03:07:58,480 --> 03:07:59,560 [INDISCERNIBLE]. 4160 03:07:59,560 --> 03:08:00,640 ESSENTIALLY WHAT DOES THE TOOL 4161 03:08:00,640 --> 03:08:01,720 TRY TO HELP? 4162 03:08:01,720 --> 03:08:16,600 HAD AND THIS IS PRIMARILY WITH 4163 03:08:16,600 --> 03:08:17,240 BIOLOGIST IN PERSPECTIVE MARKED 4164 03:08:17,240 --> 03:08:17,920 FOR IDENTIFICATION LIFE THAT'S 4165 03:08:17,920 --> 03:08:20,680 WHAT THE TOOL TRIES TO HELP YOU, 4166 03:08:20,680 --> 03:08:21,040 TOO. 4167 03:08:21,040 --> 03:08:22,560 NUMBER BE IN 2 IT CAN OOH 4168 03:08:22,560 --> 03:08:24,840 DEBTIFY THE SITE OF BINDING 4169 03:08:24,840 --> 03:08:25,880 WHICH MEANS, THESE ARE A 4170 03:08:25,880 --> 03:08:28,240 CRITICAL BINDING AND WHAT SHOULD 4171 03:08:28,240 --> 03:08:29,720 A BIOLOGIST MUTATE IN ORDER TO 4172 03:08:29,720 --> 03:08:31,920 GO FURTHER DOWN THE LINE, IT CAN 4173 03:08:31,920 --> 03:08:33,080 PREDICT THE STRENGTH OF 4174 03:08:33,080 --> 03:08:34,480 INTERACTION WHICH IS THE 4175 03:08:34,480 --> 03:08:35,920 RELATIVE AFFINITY FOR MOST 4176 03:08:35,920 --> 03:08:38,680 PURPOSES AND THEN, YOU KNOW WHAT 4177 03:08:38,680 --> 03:08:40,760 MED A POTENTIAL MECHANISM BE, 4178 03:08:40,760 --> 03:08:43,120 WOULD IT BE AN ALOE STATIC 4179 03:08:43,120 --> 03:08:44,840 MECHANISM, WHAT ARE THE ROLE OF 4180 03:08:44,840 --> 03:08:46,680 THE LENGTH OF THE CHAIN, THINGS 4181 03:08:46,680 --> 03:08:48,360 OF THAT SORT SO ALL OF DWHEES 4182 03:08:48,360 --> 03:08:50,240 CAN BE ANSWERED USING THE TOOL 4183 03:08:50,240 --> 03:08:51,560 THAT YOU DEVELOP. 4184 03:08:51,560 --> 03:08:54,160 NOW I WANT TO HOPEFULLY THE 5 4185 03:08:54,160 --> 03:08:55,480 MINUTES I HAVE, I WILL TELL YOU 4186 03:08:55,480 --> 03:08:58,680 ABOUT A VERY INTERESTING STORY 4187 03:08:58,680 --> 03:09:04,440 PRIMARILY BECAUSE IT ALERTS 4188 03:09:04,440 --> 03:09:08,200 BIOLOGIST AS WELL AS CHEMISTS TO 4189 03:09:08,200 --> 03:09:13,640 NOT ASSUME ANY [INDISCERNIBLE]. 4190 03:09:13,640 --> 03:09:16,840 AND WHICH WAS DONE WITH 4191 03:09:16,840 --> 03:09:18,880 PHYSICIAN SCIENTISTS, AND THESE 4192 03:09:18,880 --> 03:09:20,640 KINASES AND YOU KNOW THEY DO 4193 03:09:20,640 --> 03:09:22,520 THIS BY VARIOUS MECHANISMS, 4194 03:09:22,520 --> 03:09:25,400 DIRECT BINDING TO THE RTK, OR 4195 03:09:25,400 --> 03:09:27,720 BINDING TO THE LIGAND AND THEN 4196 03:09:27,720 --> 03:09:29,040 MODULATING THINGS OF THAT SORT 4197 03:09:29,040 --> 03:09:31,320 AND VARIOUS, YOU KNOW RDKs 4198 03:09:31,320 --> 03:09:43,040 HAVE BEEN, YOU KNOW STUDIED FOR 4199 03:09:43,040 --> 03:09:45,880 THIS FDFR, AND ABOUT GAG BINDING 4200 03:09:45,880 --> 03:09:48,880 TO IGFR, AND IGF 1 SYSTEM AND WE 4201 03:09:48,880 --> 03:09:50,960 KNOW IGF1 R AND 1 IS IMPORTANT 4202 03:09:50,960 --> 03:09:52,560 BOTHOT CANCER PERSPECTIVE AS 4203 03:09:52,560 --> 03:09:54,200 WELL AS FROM GNLT GROWTH 4204 03:09:54,200 --> 03:09:55,160 PERSPECTIVE AND AGING 4205 03:09:55,160 --> 03:09:56,320 PERSPECTACLIVE, THINGS OF THAT 4206 03:09:56,320 --> 03:09:58,440 SORT, THE UNIQUE THING ABOUT IGF 4207 03:09:58,440 --> 03:10:00,400 1 R IS IT'S A CONSTITTATIVE 4208 03:10:00,400 --> 03:10:03,240 DIMER AND THE WAY IN WHICH 4209 03:10:03,240 --> 03:10:05,320 ACTIVATION WORK SYSTEM THAT THE 4210 03:10:05,320 --> 03:10:07,640 TRANSMEMBRANE HAVE TO CROSS EACH 4211 03:10:07,640 --> 03:10:09,640 OTHER, BRING THE KINASE DOMAINS 4212 03:10:09,640 --> 03:10:12,280 IN THE CELL CLOTHE TOGETHER IN 4213 03:10:12,280 --> 03:10:15,040 ORDER TO ENGAGE 4214 03:10:15,040 --> 03:10:16,520 AUTOPHOSPHORYLATION. 4215 03:10:16,520 --> 03:10:21,560 NOW, I GF1, 1 MOLECULE OF IGF 1, 4216 03:10:21,560 --> 03:10:27,200 1 IGFR TO 1 IGF1 IS A DIMER SO 4217 03:10:27,200 --> 03:10:28,600 MONITOR OR 4218 03:10:28,600 --> 03:10:29,160 MERIGFWOBDERFUL--WONDERFUL, 4219 03:10:29,160 --> 03:10:31,200 WOULD ACTUALLY DO YOU KNOW 4220 03:10:31,200 --> 03:10:31,480 ACTIVATION. 4221 03:10:31,480 --> 03:10:34,320 WHAT OF THE ROLE OF HS, 4222 03:10:34,320 --> 03:10:35,240 COMPLETELY UNKNOWN BECAUSE OF 4223 03:10:35,240 --> 03:10:38,000 OUR PRIOR WORK WITH DR. PATHWAY 4224 03:10:38,000 --> 03:10:39,640 GIVES TELETHAT WE HAD KNOWN THAT 4225 03:10:39,640 --> 03:10:41,160 HS WOULD ENGAGE IN, WOULD DO 4226 03:10:41,160 --> 03:10:43,640 SOMETHING, WE ARE NOT REALLY 4227 03:10:43,640 --> 03:10:45,160 UNDERSTOOD IT FULLY. 4228 03:10:45,160 --> 03:10:47,920 SO WE ASKED A QUESTION WHETHER 4229 03:10:47,920 --> 03:10:48,600 OLIGIO SACCHARIDES CAN DO SOME 4230 03:10:48,600 --> 03:10:51,320 TRICK AND WHAT IS THE CHAIN 4231 03:10:51,320 --> 03:10:52,480 DEPENDENCE AND WE HAD 4232 03:10:52,480 --> 03:10:55,640 INVESTIGATED THIS, WE HAD 4233 03:10:55,640 --> 03:10:57,680 PUBLISHED THIS SOMETIME AGO, SO 4234 03:10:57,680 --> 03:10:59,560 I'M SIMPLY PRESENTING THE 4235 03:10:59,560 --> 03:11:00,240 COMPUTATIONAL ASPECT BECAUSE YOU 4236 03:11:00,240 --> 03:11:02,360 WANT TO SEE HOW THIS HAS HELPED 4237 03:11:02,360 --> 03:11:04,360 THE BIOLOGIST OUT BUT BASICALLY 4238 03:11:04,360 --> 03:11:08,160 IGF 1 R, THE ACTIVE DOMAIN IS 4239 03:11:08,160 --> 03:11:10,040 EXTENSIVE, CONSISTS OF L1 4240 03:11:10,040 --> 03:11:11,640 DOMAIN, CRDOMAIN AND L2 DOMAIN 4241 03:11:11,640 --> 03:11:13,320 AND ESSENTIA WILY NOTHING WAS 4242 03:11:13,320 --> 03:11:15,880 KNOWN ABOUT GAG BINDING SO WE 4243 03:11:15,880 --> 03:11:17,680 DECIDED THE ENTIRE PROTEIN INTO 4244 03:11:17,680 --> 03:11:22,960 THE BINDING SITES. 4245 03:11:22,960 --> 03:11:24,400 BS1 TO-7 AND ESSENTIALLY 4246 03:11:24,400 --> 03:11:27,200 DR. [INDISCERNIBLE] OF 4247 03:11:27,200 --> 03:11:27,880 DISCIPLINARY SACCHARIDES, 4248 03:11:27,880 --> 03:11:29,040 HEXASACCHARIDES, OCT O 4249 03:11:29,040 --> 03:11:30,720 SACCHARIDES ON THESE ARE SITES, 4250 03:11:30,720 --> 03:11:35,520 USING THE 2 FILTERS, AND AND TO 4251 03:11:35,520 --> 03:11:37,800 PUT THE WHOLE SORT OF STORY, THE 4252 03:11:37,800 --> 03:11:41,000 BOTTOM LINE OF THE STORY 4253 03:11:41,000 --> 03:11:41,840 INTEGRAL EXPERIMENTS OVER A 4254 03:11:41,840 --> 03:11:42,840 FAIRLY LONG PERIOD OF TIME AND 4255 03:11:42,840 --> 03:11:45,040 THIS IS THE UNUSUAL PART THAT WE 4256 03:11:45,040 --> 03:11:47,240 FOUND THAT IS THE PREDICTION 4257 03:11:47,240 --> 03:11:49,640 WISE THAT THE HEXASACCHARIDE 4258 03:11:49,640 --> 03:11:52,040 BINDS IN AN INTERFACE BETWEEN 4259 03:11:52,040 --> 03:11:55,880 BS2 SQUARELY FIT BS 7 IN REAL 4260 03:11:55,880 --> 03:11:57,520 LIFE ISHT PHASE BETWEEN THE L1 4261 03:11:57,520 --> 03:12:01,320 DOMAIN AND THE CR DOMAIN. 4262 03:12:01,320 --> 03:12:02,840 QUITE THE OPPOSITE, AND WHAT IT 4263 03:12:02,840 --> 03:12:05,600 ACTUALLY MEANT WAS THAT LONGER 4264 03:12:05,600 --> 03:12:06,880 OLIGIO SACCHARIDES DO NOT BIND 4265 03:12:06,880 --> 03:12:08,600 IN THIS PARTICULAR SITE. 4266 03:12:08,600 --> 03:12:11,760 THEY ACTUALLY PREFER TO BIND IN 4267 03:12:11,760 --> 03:12:13,880 A SITE COMPLETELY DIFFERENT FROM 4268 03:12:13,880 --> 03:12:14,320 THE HEXASACCHARIDE. 4269 03:12:14,320 --> 03:12:17,480 NOW THIS IS THE REVEALING 4270 03:12:17,480 --> 03:12:17,720 PORTION. 4271 03:12:17,720 --> 03:12:18,040 WHY? 4272 03:12:18,040 --> 03:12:18,920 BECAUSE GENERALLY SPEAKING WE 4273 03:12:18,920 --> 03:12:21,240 CHEMISTS AS WELL AS BIOLOGISTS 4274 03:12:21,240 --> 03:12:23,360 ASSUME THAT A SACCHARIDES BINDS 4275 03:12:23,360 --> 03:12:24,640 IN A PARTICULAR SITE, IT WILL 4276 03:12:24,640 --> 03:12:28,280 BIND WITH THE SAME SITE, THE 4277 03:12:28,280 --> 03:12:29,280 HEXASACCHARIDE BINDS WITH THE 4278 03:12:29,280 --> 03:12:31,040 HIGH AFUNNITY OF THE SAME SITE, 4279 03:12:31,040 --> 03:12:33,120 THAT IS NOT REALLY TRUE AS FAR 4280 03:12:33,120 --> 03:12:36,640 AS PREDICTION GOES, WITH CVLS, 4281 03:12:36,640 --> 03:12:38,040 THIS ALSO FITTED WELL WITH THE 4282 03:12:38,040 --> 03:12:41,240 BIOLOGY DATA WE HAD WHICH YOU 4283 03:12:41,240 --> 03:12:45,160 KNOW WE HAD PUBLISHED NOW, IN 4284 03:12:45,160 --> 03:12:47,040 TERMS OF THE STRENGTHS OF 4285 03:12:47,040 --> 03:12:48,400 TECHNIQUE, THE TECHNIQUE COULD 4286 03:12:48,400 --> 03:12:49,920 NOT ONLY PREDICT THE SITE OF 4287 03:12:49,920 --> 03:12:52,120 BINDING BUT IT COULD IDENTIFY 4288 03:12:52,120 --> 03:12:53,800 RESIDUES THAT ARE IMPORTANT FOR 4289 03:12:53,800 --> 03:12:55,600 BINDING THAT 1 CAN ACTUALLY 4290 03:12:55,600 --> 03:12:57,520 MUTATE AS TO WHAT WOULD THEIR 4291 03:12:57,520 --> 03:12:59,040 CONTRIBUTION BE, SO BRINGING THE 4292 03:12:59,040 --> 03:13:00,920 WHOLE SORT EVER STORY SHORT AND 4293 03:13:00,920 --> 03:13:02,120 I'M PRESENTING ONLY VERY SMALL 4294 03:13:02,120 --> 03:13:05,640 PIECE OF DATA TO SUPPORT THE 4295 03:13:05,640 --> 03:13:07,240 CDLS PREDICTION, THAT WE STARTED 4296 03:13:07,240 --> 03:13:12,520 YOU KNOW A BUNCH OF, 4297 03:13:12,520 --> 03:13:14,400 THERMODYNAMIC AFFINITY STUDIESS, 4298 03:13:14,400 --> 03:13:16,080 USING FLUORESCENCE AND USING 4299 03:13:16,080 --> 03:13:16,920 TECHNIQUES AND THE BOTTOM LINE 4300 03:13:16,920 --> 03:13:20,160 IS AS YOU CAN SEE IN THE PANEL 4301 03:13:20,160 --> 03:13:22,560 C, SO THE PANEL C HAS 2 LINES, 1 4302 03:13:22,560 --> 03:13:25,800 IS RED, AND 1 ISIBLY, 1 STANDS 4303 03:13:25,800 --> 03:13:29,160 HEAR HSO6, WHEN IS THE 4304 03:13:29,160 --> 03:13:31,240 HEXASACCHARIDE, THE HS12 IS THE 4305 03:13:31,240 --> 03:13:31,840 [INDISCERNIBLE] SACCHARIDE AND 4306 03:13:31,840 --> 03:13:35,080 YOU CAN SEE THAT THE AFFINITY 4307 03:13:35,080 --> 03:13:38,000 INCREASES FOR THE HEXASACCHARIDE 4308 03:13:38,000 --> 03:13:39,360 AS THE CONCENTRATION IGF1 4309 03:13:39,360 --> 03:13:43,320 INCREASES AND THIS IS TYPICALLY 4310 03:13:43,320 --> 03:13:44,640 TRUE FOR COMPETITION. 4311 03:13:44,640 --> 03:13:45,240 FOR COMPETITIVE BIND 4312 03:13:45,240 --> 03:13:47,440 THANKSGIVING IS REALLY TRUE AND 4313 03:13:47,440 --> 03:13:49,320 FOR HS12, ESSENTIALLY THIS IS NO 4314 03:13:49,320 --> 03:13:51,040 COMPETITION, THE AFFINITY DOES 4315 03:13:51,040 --> 03:13:52,240 NOT CHANGE AT ALL. 4316 03:13:52,240 --> 03:13:54,960 WHICH MEANS THAT THEA SITE OF 4317 03:13:54,960 --> 03:13:55,760 BINDING ARE COMPLETELY 4318 03:13:55,760 --> 03:13:58,160 DIFFERENT, ALTHOUGH WE ARE ONLY 4319 03:13:58,160 --> 03:13:59,640 TALKING ABOUT THE ECTODOMAIN, SO 4320 03:13:59,640 --> 03:14:01,200 THIS IS THE REVEALING PART, IT'S 4321 03:14:01,200 --> 03:14:02,240 ACTUALLY A PRINCIPLE THAT I 4322 03:14:02,240 --> 03:14:03,840 THINK WE ALL TO BE FAMILIAR 4323 03:14:03,840 --> 03:14:06,920 WITH, NOT REALLY JUSTA ASSUME 4324 03:14:06,920 --> 03:14:08,720 THAT THE OLIGO SACCHARIDES BIND 4325 03:14:08,720 --> 03:14:10,520 IN A REPET TEF MANNER AND 4326 03:14:10,520 --> 03:14:12,440 STATISTICAL INCREASE AND 4327 03:14:12,440 --> 03:14:14,720 AFFINITY ARISES FROM IN FACT IN 4328 03:14:14,720 --> 03:14:15,320 THIS PARTICULAR CASE, HS 12 4329 03:14:15,320 --> 03:14:18,200 BINDS WITH THE WEAKER AFFINITY 4330 03:14:18,200 --> 03:14:21,200 UNDER CERTAIN CONDITIONS THAN 4331 03:14:21,200 --> 03:14:24,640 HSO 6, HSO 6 BINDS TIGHTER THAN 4332 03:14:24,640 --> 03:14:27,120 12, THE LONGER SACCHARIDE BINDS 4333 03:14:27,120 --> 03:14:29,320 WEAKER AND THE SHORTER 1 BINDS. 4334 03:14:29,320 --> 03:14:30,640 NOW BRINGS THE STORY TO A CLOSE 4335 03:14:30,640 --> 03:14:32,640 AT THIS POINT, YOU KNOW WE 4336 03:14:32,640 --> 03:14:34,120 MUTATED THE 2 RESIDUES THAT WE 4337 03:14:34,120 --> 03:14:39,640 TALKED ABOUT VERY IMPORTANT 4338 03:14:39,640 --> 03:14:41,240 WHICH IS R-GENINE 1 AND 38, AND 4339 03:14:41,240 --> 03:14:43,240 WE FOUND A SIGNIFICANT INCREASE 4340 03:14:43,240 --> 03:14:44,840 IN THE AFFINITY FOR THE 4341 03:14:44,840 --> 03:14:45,440 HEXASACCHARIDE. 4342 03:14:45,440 --> 03:14:46,840 THIS MEANS THE HYPOTHESIS, THE 4343 03:14:46,840 --> 03:14:49,040 PREDICTION APPEARS TO BE 4344 03:14:49,040 --> 03:14:50,120 REASONABLY CORRECT, WHAT I'M NOT 4345 03:14:50,120 --> 03:14:52,240 SHOWING YOU IS ALL THE IMMUNO 4346 03:14:52,240 --> 03:14:52,920 FLUORESCENCE DATA THAT SUPPORT 4347 03:14:52,920 --> 03:14:54,800 THIS IS IDEA AND THE PAPER IS 4348 03:14:54,800 --> 03:14:55,680 CURRENTLY IN REVIEW. 4349 03:14:55,680 --> 03:14:56,880 ANYWAY, I THINK OVER ALL WHAT I 4350 03:14:56,880 --> 03:14:59,520 WOULD LIKE TO SAY IS THAT THIS 4351 03:14:59,520 --> 03:15:01,280 U1 AWARD HAS BEEN VERY MUCH 4352 03:15:01,280 --> 03:15:03,120 BENEFICIAL TO US, WE WOULD HAVE 4353 03:15:03,120 --> 03:15:04,080 NEVER EMBARK OFFICE OF DIVERSITY 4354 03:15:04,080 --> 03:15:05,320 DEVELOPING A WEB SERVER AND I 4355 03:15:05,320 --> 03:15:08,840 SAID THIS EARLIER, TOO, BECAUSE 4356 03:15:08,840 --> 03:15:10,840 IT IS SOMETHING THAT YOU KNOW WE 4357 03:15:10,840 --> 03:15:14,480 WOULD NOT REALLY THINK ABOUT, 4358 03:15:14,480 --> 03:15:16,360 BUT THE UO1 AWARD MADE IT 4359 03:15:16,360 --> 03:15:17,880 POSSIBLE BY HIDING APPROPRIATE 4360 03:15:17,880 --> 03:15:20,240 WEB DEVELOPERS, AND THINGS OF 4361 03:15:20,240 --> 03:15:21,800 THAT SORT. 4362 03:15:21,800 --> 03:15:23,840 THE SITE ADDRESSES AS SHOWN 4363 03:15:23,840 --> 03:15:25,040 HERE, WE HAVE STUDIED VARIOUS 4364 03:15:25,040 --> 03:15:27,160 SYSTEMS, I WOULD SAY WE HAVE NOW 4365 03:15:27,160 --> 03:15:29,600 STARTED PROBABLY DOZEN AND HALF 4366 03:15:29,600 --> 03:15:31,920 PROTEINS AND THE ALGORITHM WORKS 4367 03:15:31,920 --> 03:15:33,320 REALLY WELL ACROSS VERY, VERY 4368 03:15:33,320 --> 03:15:35,880 DIFFERENT TYPES OF PROTEINS AND 4369 03:15:35,880 --> 03:15:38,200 RECEPTORS, AND YOU KNOW SO MANY 4370 03:15:38,200 --> 03:15:41,640 OTHERS AND WE THIS WILL CONTINUE 4371 03:15:41,640 --> 03:15:41,880 FORWARD. 4372 03:15:41,880 --> 03:15:43,440 WE WOULD REALLY WELCOME OTHER 4373 03:15:43,440 --> 03:15:45,120 GRUPS WHO ASSESS THE ROLE OF 4374 03:15:45,120 --> 03:15:46,520 GAGS IN THEIR SYSTEM AS WELL AS 4375 03:15:46,520 --> 03:15:49,040 YOU KNOW IF THERE IS ANY 4376 03:15:49,040 --> 03:15:49,840 COLLABORATIVE EFFORT THAT THEY 4377 03:15:49,840 --> 03:15:53,440 WOULD LIKE TO ENGAGE IN, WE WILL 4378 03:15:53,440 --> 03:15:54,160 WELCOME THAT, TOO. 4379 03:15:54,160 --> 03:15:55,680 FINALLY, THIS IS MY GROUP. 4380 03:15:55,680 --> 03:15:56,320 IT'S A FAIRLY OLD FORMULARY 4381 03:15:56,320 --> 03:15:58,120 REFERENCE FILE BUT I WOULD LIKE 4382 03:15:58,120 --> 03:15:59,960 TO HIGHLIGHT THE SPECIAL ROLE OF 4383 03:15:59,960 --> 03:16:02,280 2 INDIVIDUALS WHO DEVELOPED THE 4384 03:16:02,280 --> 03:16:04,280 WEB SERVER AND HELPED THE 4385 03:16:04,280 --> 03:16:07,640 BIOLOGIST, THE PERSON IN RED 4386 03:16:07,640 --> 03:16:14,240 T-SHIRT ON RIGHT IS 4387 03:16:14,240 --> 03:16:17,440 [INDISCERNIBLE] AS WELL AS THE 4388 03:16:17,440 --> 03:16:19,640 YELLOW T-SHIRT, ARE THE TOO 4389 03:16:19,640 --> 03:16:20,640 COMPUTATIONAL BIOLOGIST WHO DID 4390 03:16:20,640 --> 03:16:21,880 A LOT OF WORK. 4391 03:16:21,880 --> 03:16:22,520 THANKS A LOT. 4392 03:16:22,520 --> 03:16:24,440 I KNOW I AM PREVEBTING YOU FROM 4393 03:16:24,440 --> 03:16:26,560 HAVING GOOD LUNCH BUT HOPEFULLY 4394 03:16:26,560 --> 03:16:27,680 WITH THIS YOU KNOW YOU HAVE 4395 03:16:27,680 --> 03:16:33,360 ENJOYED MY TALK AND GREATLY 4396 03:16:33,360 --> 03:16:33,680 APPRECIATE IT. 4397 03:16:33,680 --> 03:19:43,040 [ APPLAUSE ] 4398 03:19:43,040 --> 03:19:44,480 LAST STEP, THE ENZYME THAT 4399 03:19:44,480 --> 03:19:46,200 CATALYZING THE LAST STEP IN THE 4400 03:19:46,200 --> 03:19:47,840 PATHWAY, IN ADDITION TO 4401 03:19:47,840 --> 03:19:49,840 CHANGESES IN ABUNDANCE WE ALSO 4402 03:19:49,840 --> 03:19:54,680 SEE INCREASED ABUNDANCE OF THE 4403 03:19:54,680 --> 03:19:56,000 OVAL CONNECT TRANSFERASE. 4404 03:19:56,000 --> 03:19:58,800 SPEAKING OF THE ROLE OF 4405 03:19:58,800 --> 03:20:00,760 O-GlcNac IN POTENTIATING 4406 03:20:00,760 --> 03:20:03,840 TUMORIGENESIS, IN ADDITION TO 4407 03:20:03,840 --> 03:20:06,440 GFAT WITH FIVE DAY ABBREVIATED 4408 03:20:06,440 --> 03:20:08,360 TO DON REDUCES TUMOR LOAD AS 4409 03:20:08,360 --> 03:20:11,440 SHOWN IN THIS UPPER RIGHT HAND 4410 03:20:11,440 --> 03:20:12,240 PANEL. THESE DATA ARE 4411 03:20:12,240 --> 03:20:14,600 REINFORCED BY GENETIC STUDIES 4412 03:20:14,600 --> 03:20:18,320 UTILIZING A KNOCK OUT OF THE 4413 03:20:18,320 --> 03:20:20,040 O-GlcNac TRANSFERASE IN 4414 03:20:20,040 --> 03:20:21,480 LUNG TISSUE SHOWN IN THE GREEN, 4415 03:20:21,480 --> 03:20:23,720 WHICH ENHANCES TUMOR FREE 4416 03:20:23,720 --> 03:20:25,640 SURVIVAL WHEN COMPARED TO MOCK 4417 03:20:25,640 --> 03:20:27,640 TREATED ANIMAL SHOWN IN THE BLUE 4418 03:20:27,640 --> 03:20:30,360 BY ANIMALS TREATED WITH AN 4419 03:20:30,360 --> 03:20:34,640 INHIBITOR THIGH MATE G WHICH 4420 03:20:34,640 --> 03:20:36,160 ENHANCES O-GlcNac LEVELS 4421 03:20:36,160 --> 03:20:40,040 BY INHIBITING THE O-GlcNac 4422 03:20:40,040 --> 03:20:41,640 AIS. THESE MODULATING 4423 03:20:41,640 --> 03:20:43,040 O-GlcNac CELL CULTURE IN 4424 03:20:43,040 --> 03:20:45,840 VIVO IS NOT WITHOUT CHALLENGE. 4425 03:20:45,840 --> 03:20:47,320 INHIBITORS OF THE O-GlcNac 4426 03:20:47,320 --> 03:20:49,720 HAVE BEEN TREMENDOUSLY POWERFUL 4427 03:20:49,720 --> 03:20:51,240 AS HAVE MURINE MODELS THAT 4428 03:20:51,240 --> 03:20:53,800 ENABLE TO DELETE OGT AND OGA 4429 03:20:53,800 --> 03:20:56,680 FROM SPECIFIC CELLS. HOWEVER, 4430 03:20:56,680 --> 03:20:59,120 OFTEN THESE APPROACHES LACK 4431 03:20:59,120 --> 03:21:01,040 TISSUE SPECIFICITY AND CAN BE 4432 03:21:01,040 --> 03:21:02,920 OFF TARGET EFFECTS AND THIS IS 4433 03:21:02,920 --> 03:21:06,160 ESPECIALLY TRUE OF CURRENT HBPN 4434 03:21:06,160 --> 03:21:08,240 INHIBITORS. OVEREXPRESSING OGT 4435 03:21:08,240 --> 03:21:10,400 AND OGA AS WELL AS DELETING THEM 4436 03:21:10,400 --> 03:21:14,520 CAN BE LETHAL LETHAL. AND AS JEN AND 4437 03:21:14,520 --> 03:21:19,600 COLLABORATORS ARE EXPLOITING, 4438 03:21:19,600 --> 03:21:22,640 EYE BUN DANCE OF ENZYMES IS 4439 03:21:22,640 --> 03:21:23,120 ITSELF REGULATED BY 4440 03:21:23,120 --> 03:21:23,680 O-GlcNac LEVELS. THE 4441 03:21:23,680 --> 03:21:26,520 LATTER IS CRITICAL IN ADDITION 4442 03:21:26,520 --> 03:21:27,760 TO CATALYTIC ACTIVITY BOTH 4443 03:21:27,760 --> 03:21:29,760 ENZYMES ARE BELIEVED TO PLAY 4444 03:21:29,760 --> 03:21:31,520 SCAFFOLDING ROLES IN THE CELL. 4445 03:21:31,520 --> 03:21:34,320 THUS THE GOALS OF OUTCOME OUR 4446 03:21:34,320 --> 03:21:36,160 COMMON FUND SUPPORT WERE TO 4447 03:21:36,160 --> 03:21:38,280 GENERATE GENETICALLY ENCODED OGT 4448 03:21:38,280 --> 03:21:40,320 AND OGA CONSTRUCTS THAT ALLOW 4449 03:21:40,320 --> 03:21:41,720 SMALL MOLECULE REGULATION OF 4450 03:21:41,720 --> 03:21:46,000 O-GlcNac LEVELS. THE 4451 03:21:46,000 --> 03:21:48,840 APPROACH WE CHOSE RELIES ON 4452 03:21:48,840 --> 03:21:50,240 DESTABILIZATION OF THE MAIN 4453 03:21:50,240 --> 03:21:52,840 TECHNOLOGY DEVELOPED IN THE LAB 4454 03:21:52,840 --> 03:21:55,880 AT STANFORD. HERE FUSION OF 4455 03:21:55,880 --> 03:21:59,240 DESTABILIZATION DOMAIN BASED ON 4456 03:21:59,240 --> 03:22:01,880 FKB 12 OR HYDRO FOLATE REDUCTASE 4457 03:22:01,880 --> 03:22:05,080 TO PROTEIN OF INTEREST RESULTS 4458 03:22:05,080 --> 03:22:07,240 IN FUSION PROTEIN DEGRADATION. 4459 03:22:07,240 --> 03:22:08,640 THESE DESTABILIZATION DOMAINS 4460 03:22:08,640 --> 03:22:11,680 ARE MUTATED SUCH THAT BINDING TO 4461 03:22:11,680 --> 03:22:14,520 LIGAND EITHER SHIELD OR TRIMETH 4462 03:22:14,520 --> 03:22:16,400 PRONE STABILIZES THE 4463 03:22:16,400 --> 03:22:18,640 DESTABILIZATION DOMAIN AND IN 4464 03:22:18,640 --> 03:22:20,400 TURN THAT OF FUSION PROTEIN. 4465 03:22:20,400 --> 03:22:23,240 THIS APPROACH ALLOWS RAPID DOSE 4466 03:22:23,240 --> 03:22:25,800 DEPENDENT INDUCTION OF PROTEIN 4467 03:22:25,800 --> 03:22:28,320 -- INDUCTION OF PROTEIN 4468 03:22:28,320 --> 03:22:29,600 ABUNDANCE INDEPENDENT OPHIURANS 4469 03:22:29,600 --> 03:22:32,400 SUBSCRIPTIONAL AND 4470 03:22:32,400 --> 03:22:33,120 POSTTRANSCRIPTIONAL REGULATION. 4471 03:22:33,120 --> 03:22:34,640 AS TOOLS ARE GENETIC IT IS 4472 03:22:34,640 --> 03:22:37,240 POSSIBLE TO GENERATE INDUCIBLE 4473 03:22:37,240 --> 03:22:39,200 SYSTEMS ALLEVIATING OFF TARGET 4474 03:22:39,200 --> 03:22:41,480 EFFECTS OF VIRAL TRANSDUCTION, 4475 03:22:41,480 --> 03:22:43,320 TRANSIENT TRAN FECKS AND SHORT 4476 03:22:43,320 --> 03:22:45,320 HAIR PIN SHORT INTERFERON RNA 4477 03:22:45,320 --> 03:22:48,240 KNOCK DOWN. IT IS ALSO POSSIBLE 4478 03:22:48,240 --> 03:22:51,240 TO WASH THE DRUGS OUT PROVIDING 4479 03:22:51,240 --> 03:22:53,720 CRITICAL CONTROLS. EXAMPLE OF 4480 03:22:53,720 --> 03:22:57,440 DESTABILIZED OGA IS SHOWN HERE 4481 03:22:57,440 --> 03:22:59,240 IN LOWER PANEL DEMONSTRATING 4482 03:22:59,240 --> 03:23:01,040 PROBUST CHANGES IN OGA ABUNDANCE 4483 03:23:01,040 --> 03:23:02,520 IN ONLY FOUR HOURS AS WELL AS 4484 03:23:02,520 --> 03:23:05,560 CHANGES IN THE ACTIVITY OF 4485 03:23:05,560 --> 03:23:08,280 TESTIFY O-GlcNac AIS WITH 4486 03:23:08,280 --> 03:23:10,200 PROOF OF PRINCIPAL IN HAND WE 4487 03:23:10,200 --> 03:23:11,640 DEVELOPED A SUITE OF TUMORS TO 4488 03:23:11,640 --> 03:23:13,400 ALLOW RESEARCHERS TO ADDRESS A 4489 03:23:13,400 --> 03:23:14,240 BROAD RANGE OF BIOLOGICAL 4490 03:23:14,240 --> 03:23:18,160 QUESTIONS. O-GlcNacIST 4491 03:23:18,160 --> 03:23:20,000 OUR OVERALL STRATEGY IS DEPICTED 4492 03:23:20,000 --> 03:23:21,960 ON THIS SLIDE. OUR STRATEGY 4493 03:23:21,960 --> 03:23:24,200 RELIES ON USING THE GATEWAY 4494 03:23:24,200 --> 03:23:29,240 RECOMBINATION SYSTEM. WE HAVE 4495 03:23:29,240 --> 03:23:30,840 GENERATE EIGHT DESTINATION 4496 03:23:30,840 --> 03:23:32,240 VECTORS WHICH EPICODE TUNING AS 4497 03:23:32,240 --> 03:23:34,160 WELL AS -- ENCODE TUNEK AS WELL 4498 03:23:34,160 --> 03:23:37,400 AS VECTORS WHICH INCLUDE OGA OGT 4499 03:23:37,400 --> 03:23:39,400 AND ENZYME WITNESS THE HBP. USE 4500 03:23:39,400 --> 03:23:44,640 OF REBOMB BY NATION GENERATES -- 4501 03:23:44,640 --> 03:23:45,240 RECOMBINATION GENERATES OUR 4502 03:23:45,240 --> 03:23:46,760 CODING SEQUENCE INTO OUR 4503 03:23:46,760 --> 03:23:48,440 DESTINATION VECTOR. THESE ARE 4504 03:23:48,440 --> 03:23:53,880 ALL THEN SEQUENCED AND THE 4505 03:23:53,880 --> 03:23:57,680 ABILITY OF CONSTRUCTS TO BE 4506 03:23:57,680 --> 03:23:59,240 STABILIZED BY TRIMETHPRONAL 4507 03:23:59,240 --> 03:24:02,560 SHIELD AND IMPACT O-GlcNac 4508 03:24:02,560 --> 03:24:04,640 LEVELS IS 964 PLATE FORMAT. 4509 03:24:04,640 --> 03:24:07,040 SUBSEQUENTLY WE SELECT THE MOST 4510 03:24:07,040 --> 03:24:08,400 TRACTABLE CONSTRUCTS MAKING 4511 03:24:08,400 --> 03:24:10,240 STABLE CELLS AND THESE ARE THEN 4512 03:24:10,240 --> 03:24:11,440 CHARACTERIZED USING MORE 4513 03:24:11,440 --> 03:24:13,920 CLASSICAL APPROACHES. OVER THE 4514 03:24:13,920 --> 03:24:15,040 NEXT FEW SLIDES I WANT TO 4515 03:24:15,040 --> 03:24:17,640 HIGHLIGHT JUST A FEW KEY POINTS 4516 03:24:17,640 --> 03:24:18,680 ASSOCIATED WITH EACH STEP IN 4517 03:24:18,680 --> 03:24:24,800 THIS PROCESS. FIRST WE 4518 03:24:24,800 --> 03:24:25,360 DEVELOPED EIGHT DESTINATION 4519 03:24:25,360 --> 03:24:26,880 VECTORS BASED ON TWO 4520 03:24:26,880 --> 03:24:28,920 COMMERCIALLY AVAILABLE 4521 03:24:28,920 --> 03:24:31,200 DESTINATION VECTORS, THAT ENCODE 4522 03:24:31,200 --> 03:24:32,560 RECOMBINATION CASSETTE SO WHERE 4523 03:24:32,560 --> 03:24:35,000 OUR CODING SEQUENCE IS 4524 03:24:35,000 --> 03:24:37,840 INTRODUCED FUSED EITHER TO GFP 4525 03:24:37,840 --> 03:24:41,160 -- FUSED TO GFP AND V 5 EPITOPE. 4526 03:24:41,160 --> 03:24:43,200 SO IN B YOU CAN SEE THAT WE HAVE 4527 03:24:43,200 --> 03:24:46,840 INTRODUCED EITHER DI HYDRO 4528 03:24:46,840 --> 03:24:49,000 FOLATE REDUCTASE OR FKBP INSTEAD 4529 03:24:49,000 --> 03:24:52,200 OF GFP ENABLING SIMPLE TUNING. 4530 03:24:52,200 --> 03:24:54,960 IN C WE HAVE INTRODUCED DIHYDRO 4531 03:24:54,960 --> 03:24:58,040 FOLATE AND FKBP FIVE PRIME OR 4532 03:24:58,040 --> 03:25:00,880 THREE PRIME TO GFP, ALLOWING 4533 03:25:00,880 --> 03:25:04,400 BOTH TUNING OF THIS CONSTRUCT 4534 03:25:04,400 --> 03:25:06,200 BUT ALSO CRACKING OF THE 4535 03:25:06,200 --> 03:25:07,920 CONSTRUCT IN VIVO. FINALLY IN D, 4536 03:25:07,920 --> 03:25:10,400 WE HAVE INTRODUCED THE DI HYDRO 4537 03:25:10,400 --> 03:25:14,360 FOLATE REDUCTASE IN FKBP 4538 03:25:14,360 --> 03:25:16,400 DOWNSTREAM OF 2A SEQUENCE WHICH 4539 03:25:16,400 --> 03:25:19,040 IS THREE PRIMED TO GFP. THESE 4540 03:25:19,040 --> 03:25:21,840 CONSTRUCTS ARE ANTICIPATED TO 4541 03:25:21,840 --> 03:25:24,120 ALLOW TUNING WITHOUT GENERATING 4542 03:25:24,120 --> 03:25:25,920 A LONG FUSION PROTEIN. HOWEVER, 4543 03:25:25,920 --> 03:25:31,240 THE GFP CAN BE USED TO CONFIRM 4544 03:25:31,240 --> 03:25:37,040 TRANSFECTION EFFICIENCY. 4545 03:25:37,040 --> 03:25:38,080 COMPLIMENTING DESTINATION 4546 03:25:38,080 --> 03:25:40,080 VECTORS ARE 40 ENTRY VECTORS 4547 03:25:40,080 --> 03:25:41,960 ENCODING THE MAJOR VARIANTS OF 4548 03:25:41,960 --> 03:25:46,040 OGT OGA UAP AND G FAT. ALLOWING 4549 03:25:46,040 --> 03:25:48,400 RESEARCHERS TO PROBE THE ROLES 4550 03:25:48,400 --> 03:25:50,720 AND REGULATION OF EACH PROTEIN 4551 03:25:50,720 --> 03:25:53,000 VARIANT. IN ADDITION TO THE 4552 03:25:53,000 --> 03:25:56,560 DIFFERENT ISOFORMS WE HAVE ALSO 4553 03:25:56,560 --> 03:25:57,400 GENERATED CATALYTICALLY 4554 03:25:57,400 --> 03:26:00,640 INEFFICIENT OR DEAD CONSTRUCTS. 4555 03:26:00,640 --> 03:26:03,000 AS WELL AS MUTATIONS 4556 03:26:03,000 --> 03:26:05,400 CHARACTERIZED BY THE COLO LAB 4557 03:26:05,400 --> 03:26:06,920 ANTICIPATED TO INCREASE 4558 03:26:06,920 --> 03:26:07,640 UTILIZATION OF UNNATURAL 4559 03:26:07,640 --> 03:26:10,160 SUBSTRATES. FINALLY WE HAVE 4560 03:26:10,160 --> 03:26:13,040 GENERATED A MUTATION AT GFAT 4561 03:26:13,040 --> 03:26:15,400 WHICH THERE IS NO FEEDBACK 4562 03:26:15,400 --> 03:26:20,920 INHIBITION BY UDP GLCNAC. THESE 4563 03:26:20,920 --> 03:26:24,040 ARE COMBINED TO GENERATE 400 4564 03:26:24,040 --> 03:26:28,080 GLYCOTUNERS SEQUENCE VERIFIED. 4565 03:26:28,080 --> 03:26:30,080 AND THEIR ABILITY TO BE 4566 03:26:30,080 --> 03:26:32,800 STABILIZED BY SHIELD AND TRIMETH 4567 03:26:32,800 --> 03:26:35,040 PRONE HAVE BEEN CHARACTERIZED IN 4568 03:26:35,040 --> 03:26:39,280 96 WELL PLATE FORMAT. IN SUM WE 4569 03:26:39,280 --> 03:26:40,920 DEVELOPED A SUITE OF TOOLS THAT 4570 03:26:40,920 --> 03:26:42,640 HAVE NUMEROUS APPLICATIONS, WE 4571 03:26:42,640 --> 03:26:44,040 HAVE GENERATED EIGHT TUNEABLE 4572 03:26:44,040 --> 03:26:47,400 DESTINATION VECTORS, 40 GATEWAY 4573 03:26:47,400 --> 03:26:50,960 COMPLIANT OG,T OGA GFAT AND UAP 4574 03:26:50,960 --> 03:26:52,400 CONSTRUCTS AS WELL AS RANGE OF 4575 03:26:52,400 --> 03:26:54,400 USEFUL MUTATIONS. THESE HAVE 4576 03:26:54,400 --> 03:26:57,880 BEEN COMBINED TO GENERATE 400 4577 03:26:57,880 --> 03:26:59,840 GLYCOTUNERS WHICH ARE MOST OF 4578 03:26:59,840 --> 03:27:03,840 THE WAY TO CHARACTERIZATION AND 4579 03:27:03,840 --> 03:27:05,840 GENERATED A RANGE OF VALIDATED 4580 03:27:05,840 --> 03:27:07,080 PROTOCOLS FOR DETECTION OF 4581 03:27:07,080 --> 03:27:11,000 O-GlcNac IN 96 WELL PLACE, 4582 03:27:11,000 --> 03:27:12,160 LIKELY GO WE ANTICIPATE THESE 4583 03:27:12,160 --> 03:27:13,440 TOOLS CAN BE COMBINED IN 4584 03:27:13,440 --> 03:27:15,760 DIFFERENT WAYS TO ANSWER 4585 03:27:15,760 --> 03:27:21,840 DIFFERENT BIOLOGICAL QUESTIONS. 4586 03:27:21,840 --> 03:27:23,760 WE THINK TUNING OF 4587 03:27:23,760 --> 03:27:24,640 O-GlcNac LEVELS IN CELL 4588 03:27:24,640 --> 03:27:26,040 CULTURE AND XENOGRAPH MODELS 4589 03:27:26,040 --> 03:27:28,520 WILL ENABLE RESEARCHERS TO 4590 03:27:28,520 --> 03:27:30,760 RECAPITULATE DISEASE MODELS. WE 4591 03:27:30,760 --> 03:27:32,720 ALSO THINK THAT THE MUTATIONS 4592 03:27:32,720 --> 03:27:34,040 THAT HAVE BUMPED OUT ACTIVE SIDE 4593 03:27:34,040 --> 03:27:37,640 WILL ALLOW THE OR IMPROVE THE 4594 03:27:37,640 --> 03:27:39,200 INCORPORATION OF UNNATURAL 4595 03:27:39,200 --> 03:27:41,760 SUGARS, FACILITATING THE 4596 03:27:41,760 --> 03:27:43,960 IDENTIFICATION OF GLYCOSYLATED 4597 03:27:43,960 --> 03:27:45,720 PROTEINS BY CHEMISTRY, OR 4598 03:27:45,720 --> 03:27:47,160 STUDYING THE DYNAMICS OF 4599 03:27:47,160 --> 03:27:51,560 GLYCOSYLATION. PROTEINS FUSED TO 4600 03:27:51,560 --> 03:27:55,120 GFP WE ANTICIPATE CAN BE TRAPPED 4601 03:27:55,120 --> 03:27:59,240 IN SPECIFIC SUB CELLULAR LOCI. 4602 03:27:59,240 --> 03:28:01,400 USING THE GFP WE THINK THAT THE 4603 03:28:01,400 --> 03:28:03,640 DIFFERENT ISOFORMS WILL ALLOW US 4604 03:28:03,640 --> 03:28:05,720 TO ASSESS DIFFERENT ISOFORM 4605 03:28:05,720 --> 03:28:07,480 FUNCTION AS WELL AS DIFFERENT 4606 03:28:07,480 --> 03:28:09,480 POST TRANSLATIONAL MODIFICATION. 4607 03:28:09,480 --> 03:28:11,320 THE TUNING VECTORS THAT WE HAVE 4608 03:28:11,320 --> 03:28:13,280 DEVELOPED CAN BE USED FOR 4609 03:28:13,280 --> 03:28:15,400 ANYTHING IN WHICH THIS APPROACH 4610 03:28:15,400 --> 03:28:17,240 WORKS SO TYPICALLY MOST PROTEINS 4611 03:28:17,240 --> 03:28:21,160 IN THE NUCLEUS AND CYTOPLASM. 4612 03:28:21,160 --> 03:28:22,440 WITH MY LAST FEW MINUTES I WANT 4613 03:28:22,440 --> 03:28:23,840 TO TELL YOU ABOUT HOW WE HAVE 4614 03:28:23,840 --> 03:28:25,400 BEEN APPLYING THESE VECTORS TO 4615 03:28:25,400 --> 03:28:28,360 STUDY THE IMPACT OF 4616 03:28:28,360 --> 03:28:31,240 O-GlcNac ON AUTOPHAGY. 4617 03:28:31,240 --> 03:28:33,360 THAT IS A HOMEOSTATIC PROCESS 4618 03:28:33,360 --> 03:28:35,480 THAT PROMOTES CELL SURVIVAL BY 4619 03:28:35,480 --> 03:28:37,400 REMOVING PROTEIN AGGREGATES AND 4620 03:28:37,400 --> 03:28:38,560 DAMAGED ORGANELLES. WE HAVE 4621 03:28:38,560 --> 03:28:41,480 BEEN INTERESTED IN ADDRESSING 4622 03:28:41,480 --> 03:28:42,080 THE HYPOTHESIS THAT 4623 03:28:42,080 --> 03:28:43,840 O-GlcNac MAY POTENTIATE 4624 03:28:43,840 --> 03:28:46,240 AUTOPHAGY, ESPECIALLY IN THE 4625 03:28:46,240 --> 03:28:47,600 HEART REDUCING CELL AND TISSUE 4626 03:28:47,600 --> 03:28:49,680 DEATH AT TIMES OF STRESS. 4627 03:28:49,680 --> 03:28:51,720 HOWEVER, ONE CHALLENGE THAT WE 4628 03:28:51,720 --> 03:28:53,200 FACE IS THAT ALL OF THE 4629 03:28:53,200 --> 03:28:55,880 APPROACHES THAT WE HAVE USED TO 4630 03:28:55,880 --> 03:28:57,680 MODULATE O-GlcNac LEVELS 4631 03:28:57,680 --> 03:29:01,040 GENETICALLY BY SI RNA AND VIRAL 4632 03:29:01,040 --> 03:29:03,600 TRANSDUCTION, HAVE ACTIVATED 4633 03:29:03,600 --> 03:29:08,400 AUTOPHAGY IN THE CONTROLS. SO 4634 03:29:08,400 --> 03:29:11,120 THIS WAS AN AWESOME PROJECT TO 4635 03:29:11,120 --> 03:29:13,080 TEST OUT OUR DIFFERENT TUNEABLE 4636 03:29:13,080 --> 03:29:16,680 CONSTRUCTS ON. SO THIS PROJECT 4637 03:29:16,680 --> 03:29:19,480 HAS BEEN LED BY CAMEL FOY A 4638 03:29:19,480 --> 03:29:21,200 RESEARCH ASSOCIATE IN THE LAB 4639 03:29:21,200 --> 03:29:24,160 AND WHAT CANAL HAS DONE IS 4640 03:29:24,160 --> 03:29:27,120 GENERATE H 9C 2 CELLS MYOBLAST 4641 03:29:27,120 --> 03:29:29,680 HEART LAKE CELLS THAT ARE STABLY 4642 03:29:29,680 --> 03:29:31,800 TRANSFECTED WITH YELLOW 4643 03:29:31,800 --> 03:29:33,720 FLUORESCENT PROTEIN OGT OR 4644 03:29:33,720 --> 03:29:36,000 CATALYTICALLY INEFFICIENT OGT IF 4645 03:29:36,000 --> 03:29:39,400 WE FOCUS ON THIS PANEL LABELED 4646 03:29:39,400 --> 03:29:40,920 LC 3 WE ARE ASSESSING CONVERSION 4647 03:29:40,920 --> 03:29:45,200 OF LC 3 FROM ITS UNLIP DATED 4648 03:29:45,200 --> 03:29:49,400 FORM TO LIP DATED FORM LC 32 AS 4649 03:29:49,400 --> 03:29:51,240 MEASURE OF INDUCTION OF 4650 03:29:51,240 --> 03:29:53,360 AUTOPHAGY. AS WE CAN SEE IN LINE 4651 03:29:53,360 --> 03:29:57,560 3 STABILIZING OGT APPEARS TO 4652 03:29:57,560 --> 03:30:00,760 ENHANCE CONVERSION OF LC 31 TO 4653 03:30:00,760 --> 03:30:02,680 LC 32, SUGGESTING ENHANCING 4654 03:30:02,680 --> 03:30:05,560 O-GlcNac LEVELS IS 4655 03:30:05,560 --> 03:30:08,640 POTENTIATING AUTOPHAGYPHAGY. 4656 03:30:08,640 --> 03:30:10,080 CATALYTICALLY INEFFICIENT SHOWN 4657 03:30:10,080 --> 03:30:12,680 IN LANE 5 SHOWS ENEMY AT 4658 03:30:12,680 --> 03:30:14,760 INTERMEDIATE PHENOTYPE. WHEN WE 4659 03:30:14,760 --> 03:30:20,560 INHIBIT LYSOSOMALATP ACE WITH 4660 03:30:20,560 --> 03:30:21,640 BAFLOMYCIN WE SEE ACCUMULATION 4661 03:30:21,640 --> 03:30:22,280 AND THIS SUGGESTS THAT 4662 03:30:22,280 --> 03:30:23,280 O-GlcNac IS POTENTIATING 4663 03:30:23,280 --> 03:30:25,840 THE INDUCTION OF AUTOPHAGY 4664 03:30:25,840 --> 03:30:29,560 RATHER THAN BLOCKING FUSION AT 4665 03:30:29,560 --> 03:30:34,160 THE AUTOPHAGOSOME AND BLASTSOME. 4666 03:30:34,160 --> 03:30:35,320 CANALS PERFORM SIMILAR 4667 03:30:35,320 --> 03:30:37,640 EXPERIMENTS USING CELLS 4668 03:30:37,640 --> 03:30:39,840 EXPRESSING O-GlcNacASE OR 4669 03:30:39,840 --> 03:30:41,640 CAT LICKICALLY INEFFICIENT 4670 03:30:41,640 --> 03:30:43,320 O-GlcNacASE. SIMILAR TO 4671 03:30:43,320 --> 03:30:45,560 OGT STABILIZATION OF THE 4672 03:30:45,560 --> 03:30:47,120 O-GlcNacASE RESULTS IN 4673 03:30:47,120 --> 03:30:48,600 ACCUMULATION OF THIS LIPID DATED 4674 03:30:48,600 --> 03:30:52,800 FORM OF LC 3 WHEREAS 4675 03:30:52,800 --> 03:30:54,760 CATALYTICALLY INEFFICIENT 4676 03:30:54,760 --> 03:30:55,880 O-GlcNacASE PHENOCOPIES IT 4677 03:30:55,880 --> 03:30:58,240 IS YELLOW FLUORESCENT PROTEIN. 4678 03:30:58,240 --> 03:31:01,800 UNLIKE OGT TREATMENT WITH 4679 03:31:01,800 --> 03:31:02,560 BAFLOMYCIN RESULTS IN LITTLE 4680 03:31:02,560 --> 03:31:04,760 CHANGE IN AMOUNTS OF LIPIDATED 4681 03:31:04,760 --> 03:31:08,480 FORM OF LC 3 IN CELLS WHERE THE 4682 03:31:08,480 --> 03:31:09,040 O-GlcNacASE HAS BEEN 4683 03:31:09,040 --> 03:31:11,920 STABILIZED. WHAT THESE DATA 4684 03:31:11,920 --> 03:31:14,120 SUGGEST IS OVER EXPRESSION OF 4685 03:31:14,120 --> 03:31:15,240 O-GlcNacASE IS BLOCKING 4686 03:31:15,240 --> 03:31:18,240 FUSION OF THE AUTOPHAGOSOME AND 4687 03:31:18,240 --> 03:31:20,840 LYSOSOME RATHER THAN 4688 03:31:20,840 --> 03:31:23,280 POTENTIATING AUTOPHAGY. SADLY I 4689 03:31:23,280 --> 03:31:25,280 DON'T HAVE TIME TO TELL YOU THE 4690 03:31:25,280 --> 03:31:27,120 REST OF CANAL'S STORY TODAY BUT 4691 03:31:27,120 --> 03:31:30,120 I DID WANT TO SUM WHAT HE 4692 03:31:30,120 --> 03:31:32,880 COMPLETED UP. HE'S DEMONSTRATED 4693 03:31:32,880 --> 03:31:34,760 THAT AUGMENTING O-GlcNac 4694 03:31:34,760 --> 03:31:37,040 LEVELS IN VIVO AND VITRO USING 4695 03:31:37,040 --> 03:31:41,600 GENETIC AND FARM O COLLAGECAL 4696 03:31:41,600 --> 03:31:46,080 APPROACHES INCREASECREASES AUTOPHAGY 4697 03:31:46,080 --> 03:31:47,360 MARKERS AND THIS IS DEPENDENTS 4698 03:31:47,360 --> 03:31:51,800 ON AMPK. ELEVATED O-GlcNac 4699 03:31:51,800 --> 03:31:55,840 ACTIVATES AMPK AND ULK 1, 4700 03:31:55,840 --> 03:31:57,840 AUTOPHAGY BLOCKS ABILITY TO 4701 03:31:57,840 --> 03:31:59,240 O-GlcNac TO MEDIATE 4702 03:31:59,240 --> 03:32:01,800 CYTOPROTECTION. AMPK IS 4703 03:32:01,800 --> 03:32:03,880 GLYCOSYLATED IN RESPONSE TO 4704 03:32:03,880 --> 03:32:05,040 OXIDATIVE STRESS AND NOW MAPPED 4705 03:32:05,040 --> 03:32:07,120 MORE THAN 12 SITES OF 4706 03:32:07,120 --> 03:32:12,640 GLYCOSYLATION IN KEY REGULATORY 4707 03:32:12,640 --> 03:32:14,840 MODULES OF AMP KINASE. THANKS 4708 03:32:14,840 --> 03:32:16,560 AGAIN TO THE COMMON FUND, IN 4709 03:32:16,560 --> 03:32:17,840 SUMMING UP HERE HOW WE HAVE 4710 03:32:17,840 --> 03:32:20,480 SHARED OUR REAGENTS, HOW WE 4711 03:32:20,480 --> 03:32:22,240 PRESENTED THIS WORK IN A RANGE 4712 03:32:22,240 --> 03:32:24,960 OF DIFFERENT SEMINARS AND 4713 03:32:24,960 --> 03:32:25,840 WORKSHOPS, AND THAT WE HAVE 4714 03:32:25,840 --> 03:32:28,440 TRIED TO VALIDATE AND 4715 03:32:28,440 --> 03:32:29,560 DISSEMINATE OUR TOOLS. THANK YOU 4716 03:32:29,560 --> 03:32:37,400 FOR YOUR ATTENTION. 4717 03:32:37,400 --> 03:32:39,840 >> OUR NEXT SPEAKER ON THIS 4718 03:32:39,840 --> 03:32:41,480 SERIES ON O-GlcNac IS DR. 4719 03:32:41,480 --> 03:32:53,040 JENNIFER KOHLER. 4720 03:32:53,040 --> 03:32:55,920 >> GOOD AFTERNOON, EVERYBODY. AS 4721 03:32:55,920 --> 03:32:58,800 I START OFF I WOULD LIKE TO 4722 03:32:58,800 --> 03:33:00,400 EXPRESS MY GRATITUDE FOR THE 4723 03:33:00,400 --> 03:33:01,720 OPPORTUNITY TO BE PART OF THE 4724 03:33:01,720 --> 03:33:04,280 COMMON FUND INITIATIVE. IT'S 4725 03:33:04,280 --> 03:33:08,000 BEEN A PLEASURE TO BE PART OF 4726 03:33:08,000 --> 03:33:09,480 THIS EFFORT MUCH THERE THAN MY 4727 03:33:09,480 --> 03:33:10,840 OWN RESEARCH GROUP AND HAVE A 4728 03:33:10,840 --> 03:33:12,440 FRONT ROW SEAT TO DEVELOPMENT OF 4729 03:33:12,440 --> 03:33:13,400 NEW TOOLS, ALL NEW TOOLS THAT 4730 03:33:13,400 --> 03:33:14,840 ARE GOING TO PAVE THE WAY FOR 4731 03:33:14,840 --> 03:33:16,040 THE FUTURE OF GLYCOSCIENCE 4732 03:33:16,040 --> 03:33:17,840 RESEARCH. 4733 03:33:17,840 --> 03:33:21,080 I WOULD LIKE TO THANK PAM, 4734 03:33:21,080 --> 03:33:23,240 AMANDA AND CARL FOR LEADING THE 4735 03:33:23,240 --> 03:33:24,840 WAY AND ALSO THANK CATHERINE 4736 03:33:24,840 --> 03:33:28,240 GRIMES FOR HER LEADERSHIP ON THE 4737 03:33:28,240 --> 03:33:31,200 TOOLS SUBGROUP. FOR OUR COMMON 4738 03:33:31,200 --> 03:33:33,080 FUND PROJECT WE -- I DECIDED 4739 03:33:33,080 --> 03:33:35,040 THAT IT WOULD BE A GREAT 4740 03:33:35,040 --> 03:33:36,400 OPPORTUNITY TO ENGAGE SEVERAL OF 4741 03:33:36,400 --> 03:33:38,000 MY COLLEAGUES FROM UT SOUTH 4742 03:33:38,000 --> 03:33:40,280 WESTERN WHO ARE NOT NECESSARILY 4743 03:33:40,280 --> 03:33:44,440 INTERESTED IN GLYCOSCIENCE BUT 4744 03:33:44,440 --> 03:33:47,240 HAVE EXPERTISE THAT ENABLES 4745 03:33:47,240 --> 03:33:48,400 GLYCOSCIENCE TOOLS SO THE WORK I 4746 03:33:48,400 --> 03:33:51,920 WILL TELL YOU ABOUT INCLUDES 4747 03:33:51,920 --> 03:33:53,560 COLLABORATIONS WITH MY 4748 03:33:53,560 --> 03:33:54,440 COLLEAGUES NICK CONRAD 4749 03:33:54,440 --> 03:33:56,960 DEPARTMENT OF MICROBIOLOGY, 4750 03:33:56,960 --> 03:33:58,840 UTTAM TAMBAR FROM SAME 4751 03:33:58,840 --> 03:34:00,240 DEPARTMENT AS ME BIOCHEMISTRY 4752 03:34:00,240 --> 03:34:05,920 AND LUKASZ JOACHIMIAK FROM THE 4753 03:34:05,920 --> 03:34:11,520 ALZHEIMER'S CENTER. THANK YOU. 4754 03:34:11,520 --> 03:34:12,880 SO THE FIRST THING I WILL TALK 4755 03:34:12,880 --> 03:34:15,840 TO YOU ABOUT IS THE PROJECT THAT 4756 03:34:15,840 --> 03:34:17,000 WAS CONDUCTED IN NICK'S LAB AND 4757 03:34:17,000 --> 03:34:21,240 THIS IS AIMED AT DEVELOPING THE 4758 03:34:21,240 --> 03:34:22,800 PORTER TO TELL US SOMETHING 4759 03:34:22,800 --> 03:34:23,840 ABOUT O-GlcNac LEVELS 4760 03:34:23,840 --> 03:34:27,240 INSIDE CELLS OR POTENTIALLY 4761 03:34:27,240 --> 03:34:29,640 INSIDE ORGANISMS. THIS REPORTER 4762 03:34:29,640 --> 03:34:31,640 WAS BASED ON OBSERVATION THAT 4763 03:34:31,640 --> 03:34:33,800 NICK CONRAD'S LAB MADE A FEW 4764 03:34:33,800 --> 03:34:36,240 YEARS AGO REGARDING THE SPLICING 4765 03:34:36,240 --> 03:34:40,200 OF THE TRANSCRIPT OF OGT, 4766 03:34:40,200 --> 03:34:41,320 PROTEIN THAT TRANSCRIPT THAT 4767 03:34:41,320 --> 03:34:43,440 ENCODES OGT PROTEIN, 4768 03:34:43,440 --> 03:34:44,280 O-GlcNac TRANSFERASE. AND 4769 03:34:44,280 --> 03:34:52,440 IN THIS CASE'S LAB IIN THISNICK'S LAB IS I NTERESTED IN 4770 03:34:52,440 --> 03:34:55,320 SPLICING AND DETAIN INTRON SOP 4771 03:34:55,320 --> 03:34:56,320 WHERE INTRONS ARE NOT 4772 03:34:56,320 --> 03:34:57,720 EFFICIENTLY SPLICED OUT OR 4773 03:34:57,720 --> 03:35:00,240 REGULATED -- REGULATORY 4774 03:35:00,240 --> 03:35:01,640 MECHANISM FOR SPLICING, THEY 4775 03:35:01,640 --> 03:35:04,000 DISCOVERED ONE INTRON IN THE OTT 4776 03:35:04,000 --> 03:35:06,080 TRANSCRIPT WAS SPLICED OUT IN 4777 03:35:06,080 --> 03:35:08,200 O-GlcNac DEPENDENT WAY SO 4778 03:35:08,200 --> 03:35:09,080 UNDER CONDITIONS OF LOW 4779 03:35:09,080 --> 03:35:12,240 O-GlcNac WITHIN THE CELL, 4780 03:35:12,240 --> 03:35:15,080 THIS TRANSCRIPT HOE SUFFICIENTLY 4781 03:35:15,080 --> 03:35:16,960 SPLICED LEADING TO OGT 4782 03:35:16,960 --> 03:35:18,920 TRANSCRIPT AND PROTEIN WHICH 4783 03:35:18,920 --> 03:35:20,240 ALLOWS INSTALLATIONS OF 4784 03:35:20,240 --> 03:35:21,960 ADDITIONAL O-GlcNac TO 4785 03:35:21,960 --> 03:35:22,400 RESTORE HOMEOSTASIS 4786 03:35:22,400 --> 03:35:23,680 O-GlcNac LEVELS, HOWEVER, 4787 03:35:23,680 --> 03:35:26,280 UNDER CONDITIONS OF HIGH 4788 03:35:26,280 --> 03:35:27,840 O-GlcNac, THIS INTRON IS 4789 03:35:27,840 --> 03:35:29,240 RETAINED AND THE TRANSCRIPT IS 4790 03:35:29,240 --> 03:35:30,840 DEGRADED AND THERE'S LESS OGT 4791 03:35:30,840 --> 03:35:35,640 AVAILABLE. SO NICK'S LAB THOUGHT 4792 03:35:35,640 --> 03:35:39,240 IT WOULD BE POSSIBLE TO THEN USE 4793 03:35:39,240 --> 03:35:40,440 THAT SAME INTRON RETENTION 4794 03:35:40,440 --> 03:35:43,000 MECHANISM TO GENERATE 4795 03:35:43,000 --> 03:35:43,760 O-GlcNac RESPONSIVE 4796 03:35:43,760 --> 03:35:45,520 REPORTERS AND THE IDEA HERE WAS 4797 03:35:45,520 --> 03:35:49,320 TO INCORPORATE THE GF -- GENE 4798 03:35:49,320 --> 03:35:51,840 ENCODING GFP ON TO THE SAME 4799 03:35:51,840 --> 03:35:53,520 TRANSCRIPT AND SO IN THIS WAY 4800 03:35:53,520 --> 03:35:56,160 THE PRODUCTION OF FLUORESCENT 4801 03:35:56,160 --> 03:35:58,040 PROTEIN WOULD NOW BE SENSITIVE 4802 03:35:58,040 --> 03:36:00,760 TO O-GlcNac LEVELS. SO 4803 03:36:00,760 --> 03:36:02,520 THEY DESIGNED A CONSTRUCT TO DO 4804 03:36:02,520 --> 03:36:04,960 THIS SO UNDER CONDITIONS OF LOW 4805 03:36:04,960 --> 03:36:06,240 O-GlcNac THIS TRANSCRIPT 4806 03:36:06,240 --> 03:36:08,640 IS SUFFICIENTLY PROCESSED, 4807 03:36:08,640 --> 03:36:09,640 PRODUCING GFP AND UNDER 4808 03:36:09,640 --> 03:36:10,640 CONDITIONS OF HIGH 4809 03:36:10,640 --> 03:36:11,560 O-GlcNac THE TRANSCRIPT IS 4810 03:36:11,560 --> 03:36:14,040 DEGRADED AND THERE'S LESS GFP. 4811 03:36:14,040 --> 03:36:17,400 SO THE LEVELS IS INVERSELY 4812 03:36:17,400 --> 03:36:18,640 PROPORTIONAL TO THE LEVEL OF 4813 03:36:18,640 --> 03:36:22,400 O-GlcNac. IN THE CELLS. 4814 03:36:22,400 --> 03:36:24,360 THEY DEVELOP THIS CONSTRUCT 4815 03:36:24,360 --> 03:36:25,680 EXPRESSED IT SABLELY AND 4816 03:36:25,680 --> 03:36:27,640 MAMMALIAN CELLS AND SHOW INDEED 4817 03:36:27,640 --> 03:36:29,640 IT IS RESPONSIVE TO 4818 03:36:29,640 --> 03:36:30,280 PERTURBATIONS THAT CHANGE 4819 03:36:30,280 --> 03:36:31,320 O-GlcNac LEVELS IN CELLS. 4820 03:36:31,320 --> 03:36:34,440 THIS IS SHOWN BY WESTERN BLOT 4821 03:36:34,440 --> 03:36:35,840 USING ANTIBODY AGAINST 4822 03:36:35,840 --> 03:36:38,640 O-GlcNac MODIFICATION AND 4823 03:36:38,640 --> 03:36:40,680 ALSO BY FLOW CYTOMETRY WHERE WE 4824 03:36:40,680 --> 03:36:44,240 CAN SEE THE LEVELS OF THE GFP 4825 03:36:44,240 --> 03:36:46,000 CHANGING IN RESPONSE TO 4826 03:36:46,000 --> 03:36:50,000 INHIBITORS SO THIAMET G INHIBITS 4827 03:36:50,000 --> 03:36:50,960 OGA AND RAISES O-GlcNac 4828 03:36:50,960 --> 03:36:53,680 LEVELS AND WE SEE LESS GFP 4829 03:36:53,680 --> 03:36:56,360 FLUORESCENCE BY FLOW CYTOMETRY, 4830 03:36:56,360 --> 03:37:00,240 WHEREAS OSMI 1 INHIBITS OGT 4831 03:37:00,240 --> 03:37:00,640 RESULTING IN LOYER 4832 03:37:00,640 --> 03:37:01,600 O-GlcNac LEVELS AND HIGHER 4833 03:37:01,600 --> 03:37:06,640 FLUORESCENCE. NICK'S LAB ALSO 4834 03:37:06,640 --> 03:37:08,160 VALIDATED THIS BY MICROSCOPY 4835 03:37:08,160 --> 03:37:12,160 WHICH IS SHOWN HERE, AND IN 4836 03:37:12,160 --> 03:37:13,720 WHERE THEY KNOCK DOWN OGT 4837 03:37:13,720 --> 03:37:15,920 TRANSCRIPT USING SI RNA AND YOU 4838 03:37:15,920 --> 03:37:19,000 CAN SEE AN INCREASE IN THE 4839 03:37:19,000 --> 03:37:20,640 FLUORESCENCE BY MICROSCOPY AND 4840 03:37:20,640 --> 03:37:23,080 FLOW CYTOMETRY. SO THIS TOOL IS 4841 03:37:23,080 --> 03:37:24,360 KIND OF READY TO BE USED IN 4842 03:37:24,360 --> 03:37:27,240 DIFFERENT WAYS AND SO YOU CAN 4843 03:37:27,240 --> 03:37:28,760 IMAGINE YOU MIGHT DO SOME 4844 03:37:28,760 --> 03:37:30,760 PERTURBATION TO THE CELL AND 4845 03:37:30,760 --> 03:37:32,320 WANT TO KNOW IF THERE IS CHANGE 4846 03:37:32,320 --> 03:37:33,600 IN O-GlcNac LEVELS AND ONE 4847 03:37:33,600 --> 03:37:34,840 COULD USE THIS REPORTER IN THIS 4848 03:37:34,840 --> 03:37:37,320 WAY. NICK'S LAB IS USING THIS 4849 03:37:37,320 --> 03:37:40,560 TO STUDY THE SPLICING OF OGT 4850 03:37:40,560 --> 03:37:42,640 TRANSCRIPT AND THEY INTRODUCED 4851 03:37:42,640 --> 03:37:44,040 THIS REPORTER, AND USED A 4852 03:37:44,040 --> 03:37:45,920 CRISPER LIBRARY AND NOW LOOKING 4853 03:37:45,920 --> 03:37:49,680 -- THEY FOUND WHICH GENES MODIFY 4854 03:37:49,680 --> 03:37:53,720 THIS RESPONSE. SO THEY TRYING TO 4855 03:37:53,720 --> 03:37:55,840 VALIDATE AND DETERMINE EXACTLY 4856 03:37:55,840 --> 03:37:57,160 WHICH SPLICE FACTORS ARE 4857 03:37:57,160 --> 03:37:59,600 INVOLVED IN THE SENSING OF 4858 03:37:59,600 --> 03:38:02,240 O-GlcNac TO REGULATE THE 4859 03:38:02,240 --> 03:38:04,360 SPLICING OF OGT TRANSCRIPT. THEY 4860 03:38:04,360 --> 03:38:06,160 ARE ALSO PRODUCING ADDITIONAL 4861 03:38:06,160 --> 03:38:08,920 REPORTER IN WHICH LUCIFERASE IS 4862 03:38:08,920 --> 03:38:10,920 PRODUCED FROM GFP AND ONE CAN 4863 03:38:10,920 --> 03:38:12,560 IMAGINE USING THAT IN ANIMALS. 4864 03:38:12,560 --> 03:38:14,040 SO IF PEOPLE ARE INTERESTED IN 4865 03:38:14,040 --> 03:38:16,040 USING THIS THEY ARE WELCOME TO 4866 03:38:16,040 --> 03:38:17,600 CONTACT NICK OR ME BECAUSE WE 4867 03:38:17,600 --> 03:38:18,720 THINK THIS REPORTER COULD BE 4868 03:38:18,720 --> 03:38:21,600 USED IN A LOT OF WAYS. ALL 4869 03:38:21,600 --> 03:38:23,040 RIGHT. NEXT KIND OF PARTS OF OUR 4870 03:38:23,040 --> 03:38:24,840 PROJECT WERE FOCUSED ON 4871 03:38:24,840 --> 03:38:26,400 UNDERSTANDING THE INTERACTION 4872 03:38:26,400 --> 03:38:28,840 PARTNERS OF GLYCOSYLATED 4873 03:38:28,840 --> 03:38:31,440 MOLECULES AND THIS CARTOON HERE 4874 03:38:31,440 --> 03:38:32,600 GIVES THE BIG PICTURE WHAT WE 4875 03:38:32,600 --> 03:38:34,240 ARE THINKING ABOUT. AND THE IDEA 4876 03:38:34,240 --> 03:38:36,880 IS THAT PROTEINS THAT ARE 4877 03:38:36,880 --> 03:38:39,240 GLYCOSYLATED IN SPECIFICALLY IN 4878 03:38:39,240 --> 03:38:45,680 THIS CASE O-GlcNac LATED 4879 03:38:45,680 --> 03:38:47,640 YOU CAN SEE THIS IN TERMS OF 4880 03:38:47,640 --> 03:38:49,440 PERHAPS WHERE THEY LOCALIZE IN 4881 03:38:49,440 --> 03:38:51,160 THE CELLS OR BINDING PARTNERS OR 4882 03:38:51,160 --> 03:38:53,440 OTHER PROTEINS THAT ARE NEARBY, 4883 03:38:53,440 --> 03:38:54,240 ONE CAN IMAGINE YOU CAN LEARN 4884 03:38:54,240 --> 03:38:55,680 ABOUT THE PROTEINS THAT THEY 4885 03:38:55,680 --> 03:38:56,960 HANG OUT WITH YOU LEARN 4886 03:38:56,960 --> 03:38:58,320 SOMETHING ABOUT WHAT THEY ARE 4887 03:38:58,320 --> 03:39:00,240 DOING. SO THAT WAS THE IDEA 4888 03:39:00,240 --> 03:39:02,040 BEHIND THIS. A FEW YEARS AGO WE 4889 03:39:02,040 --> 03:39:03,640 DECIDED TO APPROACH THIS PROBLEM 4890 03:39:03,640 --> 03:39:05,640 BY INCORPORATING A PHOTO 4891 03:39:05,640 --> 03:39:07,320 ACTIVATABLE CROSS LINKER ON TO 4892 03:39:07,320 --> 03:39:09,840 THE O-GlcNac LEVELS IN 4893 03:39:09,840 --> 03:39:13,200 LIVING CELLS. . WE DID THIS BY 4894 03:39:13,200 --> 03:39:14,280 -- I WILL DO WITHOUT THE 4895 03:39:14,280 --> 03:39:17,640 POINTER. THE WAY WE DID THIS WAS 4896 03:39:17,640 --> 03:39:20,040 BY PRODUCING THE COMPOUNDS SHOWN 4897 03:39:20,040 --> 03:39:25,960 IN THE UPPER LEFT DIAZIRINE 4898 03:39:25,960 --> 03:39:28,080 PHOTOCROSS LINKER GLOMERULAR 4899 03:39:28,080 --> 03:39:29,720 ANALOG AND PROTECTING GROUPS ARE 4900 03:39:29,720 --> 03:39:31,240 IN PURPLE. THESE PROTECTING 4901 03:39:31,240 --> 03:39:33,560 GROUPS ALLOWED US TO ENTER THE 4902 03:39:33,560 --> 03:39:36,880 CELL, REMOVE THE INTRACELLULARLY 4903 03:39:36,880 --> 03:39:39,840 REVEALING THE GLCNDAZ 1 4904 03:39:39,840 --> 03:39:42,240 PHOSPHATE. TO ACTIVATE THIS IN 4905 03:39:42,240 --> 03:39:44,440 UDP SUGAR IN CELLS WE IMMEDIATED 4906 03:39:44,440 --> 03:39:46,480 TO INTRODUCE MUTANT ENZYME 4907 03:39:46,480 --> 03:39:47,960 NATASHA POINTED OUT IN A EARLIER 4908 03:39:47,960 --> 03:39:51,120 TALK SO A MUTATION TO THE 4909 03:39:51,120 --> 03:39:51,640 O-GlcNac PHOSPHORYLATES 4910 03:39:51,640 --> 03:39:54,920 THAT ALLOWS OUTS TO ACCEPT THE 4911 03:39:54,920 --> 03:39:56,600 DIAZIRINE MODIFIED COMPOUND. 4912 03:39:56,600 --> 03:40:02,640 ONCE CELLS MAKE THIS MODIFIED 4913 03:40:02,640 --> 03:40:05,440 UDP QUICK KNACK ANALOG WITH 4914 03:40:05,440 --> 03:40:08,840 KAIZARINE IT IS A SUBSTRATE AND 4915 03:40:08,840 --> 03:40:09,520 CAN CROSS LINK O-GlcNac 4916 03:40:09,520 --> 03:40:10,760 PROTEINS TO THEIR NEIGHBORS 4917 03:40:10,760 --> 03:40:12,040 WITHIN LIVING CELLS. SO IN OUR 4918 03:40:12,040 --> 03:40:14,960 COMMON FUND PROJECT WE AIM TO 4919 03:40:14,960 --> 03:40:16,640 IMPROVE AND EXPAND THIS 4920 03:40:16,640 --> 03:40:17,600 TECHNOLOGY AND ONE THING THAT 4921 03:40:17,600 --> 03:40:19,400 WAS A PROBLEM REALLY WAS MAKING 4922 03:40:19,400 --> 03:40:21,040 THAT MOLECULE SHOWN AT THE UPPER 4923 03:40:21,040 --> 03:40:22,800 LEFT AND SO WE HAD A STRATEGY TO 4924 03:40:22,800 --> 03:40:26,080 MAKE IT BUT IT WAS I WOULD SAY 4925 03:40:26,080 --> 03:40:27,040 QUITE UNRELIABLE AND WE KNEW 4926 03:40:27,040 --> 03:40:29,440 FROM OTHERS AS WELL MAKING THIS 4927 03:40:29,440 --> 03:40:33,120 PROTECTIVE PHOSPHOSUGAR WE SAW 4928 03:40:33,120 --> 03:40:35,640 VARIABLE YIELDS BASICALLY. AND 4929 03:40:35,640 --> 03:40:38,760 SO MY COLLEAGUE UTTON TAMBAR 4930 03:40:38,760 --> 03:40:39,560 DIVIDESSED A MORE RELIABLE 4931 03:40:39,560 --> 03:40:41,920 APPROACH TO GET TO THIS MOLECULE 4932 03:40:41,920 --> 03:40:44,360 WHICH WE USE COMMERCIAL REAGENT 4933 03:40:44,360 --> 03:40:47,480 SHOWN IN UPPER LEFT IN RED TO 4934 03:40:47,480 --> 03:40:50,640 MAKE THE INITIAL PHOSPHOSUGAR 4935 03:40:50,640 --> 03:40:53,640 AND THEN EXCHANGE IN THE DIASTE 4936 03:40:53,640 --> 03:40:55,520 ARE PROTECTING GROUPS ON 4937 03:40:55,520 --> 03:40:56,840 PHOSPHATE AND FINAL OXIDATION 4938 03:40:56,840 --> 03:40:59,920 STEP. SO THIS GRAPH TO THE 4939 03:40:59,920 --> 03:41:02,400 COMPOUND IS MORE RELIABLE AND 4940 03:41:02,400 --> 03:41:04,480 MORE SCALABLE AND ALLOWS ACCESS 4941 03:41:04,480 --> 03:41:05,680 TO BOTH ORIGINAL MOLECULE WE 4942 03:41:05,680 --> 03:41:07,080 HAVE BEEN USING QUITE SOME TIME 4943 03:41:07,080 --> 03:41:10,520 AS WELL AS ADDITIONAL ANALOGS. 4944 03:41:10,520 --> 03:41:16,480 AND SO WE PREPARED THIS GLCDAZ 1 4945 03:41:16,480 --> 03:41:18,960 PHOSPHATE BUT THE GALNAC AS WELL 4946 03:41:18,960 --> 03:41:23,200 AS OTHER ANALOGS WE REPLACE 4947 03:41:23,200 --> 03:41:24,280 DIAZIRINE WITH DIFFERENT 4948 03:41:24,280 --> 03:41:25,640 FUNCTIONAL GROUPS SO WE CONTINUE 4949 03:41:25,640 --> 03:41:26,880 TOE VALUE WAIT ADDITIONAL 4950 03:41:26,880 --> 03:41:29,680 ANALOGS. -- EVALUATE THE 4951 03:41:29,680 --> 03:41:31,040 ADDITIONAL ANALOGS. THE OTHER 4952 03:41:31,040 --> 03:41:33,400 WAY TO EXPAND THE TECHNOLOGY IS 4953 03:41:33,400 --> 03:41:35,920 TESTING WHETHER THE PHOTOCROSS 4954 03:41:35,920 --> 03:41:38,040 LINKING GLCNAC WAS 4955 03:41:38,040 --> 03:41:39,400 PHOSPHORYLATED INTO 4956 03:41:39,400 --> 03:41:41,160 GLYCOCONJUGATES. THIS WAS DONE 4957 03:41:41,160 --> 03:41:42,440 IN COLLABORATION WITH SEVERAL -- 4958 03:41:42,440 --> 03:41:46,640 TWO GROUPS, AT THE CCRC, 4959 03:41:46,640 --> 03:41:47,680 (INAUDIBLE) GROUP WITH MASS 4960 03:41:47,680 --> 03:41:52,280 SPECTROMETRY ANALYSIS AND KELLY 4961 03:41:52,280 --> 03:41:53,200 KELLY'S GOSH WITH IN VITRO 4962 03:41:53,200 --> 03:41:54,920 GLCNAC TRANSFERASE ACTIVITIES 4963 03:41:54,920 --> 03:41:57,120 AND THEY DID WORK SUMMARIZED IN 4964 03:41:57,120 --> 03:41:58,640 THIS ONE SLIDE THAT ALLOWED US 4965 03:41:58,640 --> 03:42:05,280 TO CHARACTERIZE HOW THELCNGZAZ 4966 03:42:05,280 --> 03:42:07,760 WAS INCORPORATED INTO N LINKED 4967 03:42:07,760 --> 03:42:11,440 GLYCANS. WE ONLY SAW ONE 4968 03:42:11,440 --> 03:42:12,800 INCORPORATED PER LINK GLYCAN, IT 4969 03:42:12,800 --> 03:42:15,320 WAS KNOCKED DOWN IN CORE 4970 03:42:15,320 --> 03:42:16,720 POSITIONS, IT WAS WE BELIEVE 4971 03:42:16,720 --> 03:42:19,720 LIKELY INCORPORATED BY NGAP 2, 4972 03:42:19,720 --> 03:42:22,280 POTENTIALLY ALSO INCORPORATED BY 4973 03:42:22,280 --> 03:42:26,040 NGAP 5 AND IMPORTANTLY THESE 4974 03:42:26,040 --> 03:42:27,880 GLCNDAZ INCORPORATED EXTENDED 4975 03:42:27,880 --> 03:42:30,600 WITH GAU THAT ALLOWS FORMATION 4976 03:42:30,600 --> 03:42:31,560 OF MATURE GLYCANS AND LINK 4977 03:42:31,560 --> 03:42:35,480 GLYCANS. SO ONCE WE KNEW THAT 4978 03:42:35,480 --> 03:42:38,080 GLCNDAZ IS GLYCOSYLATED INTO 4979 03:42:38,080 --> 03:42:40,080 GLYCANS IN MY LAB USED THIS TOOL 4980 03:42:40,080 --> 03:42:42,560 TO LOOK AT N LINK GLYCAN BINDING 4981 03:42:42,560 --> 03:42:46,840 INTERACTIONS AND SPECIFICALLY WE 4982 03:42:46,840 --> 03:42:48,520 USED GALECTIN 1 AND WE ADDED 4983 03:42:48,520 --> 03:42:50,160 THIS EXTRA CELLULARLY TO CELLS 4984 03:42:50,160 --> 03:42:52,400 THAT WERE MAKING THESE MODIFIED 4985 03:42:52,400 --> 03:42:53,840 IN LINK GLYCANS WITH CROSS 4986 03:42:53,840 --> 03:42:57,600 LINKER AND WE COULD OBSERVE THEN 4987 03:42:57,600 --> 03:43:00,720 WITH UV RADIATION OBSERVE CROSS 4988 03:43:00,720 --> 03:43:04,360 LINKING OF GALECTIN 1 TO CELL 4989 03:43:04,360 --> 03:43:07,240 SURFACE GLYCOPROTEINS P AND BY 4990 03:43:07,240 --> 03:43:09,160 USING BIATEN LATED FORM YOU CAN 4991 03:43:09,160 --> 03:43:10,600 PULL OUT PROTEINS AND IDENTIFY 4992 03:43:10,600 --> 03:43:12,800 BY MASS SPEC AND FOUND INDEED 4993 03:43:12,800 --> 03:43:15,440 MANY ARE ANNOTATED AS N LINK 4994 03:43:15,440 --> 03:43:17,280 GLYCOPROTEINS AND SOME KNOWN TO 4995 03:43:17,280 --> 03:43:20,800 BE BINDING PARTNERS OF GALECTIN 4996 03:43:20,800 --> 03:43:22,440 1 PREVIOUSLY REPORTED IN THE 4997 03:43:22,440 --> 03:43:23,560 LITERATURE. SO IN FACT WE DID 4998 03:43:23,560 --> 03:43:26,040 THIS ANALYSIS IN CD4 CELLS AND 4999 03:43:26,040 --> 03:43:29,680 QUITE SOME TIME AGO IN THE ' 5000 03:43:29,680 --> 03:43:32,880 '90s THERE WAS A REPORT THAT 5001 03:43:32,880 --> 03:43:35,280 GALECTIN 1 WAS USED FOR AFFINITY 5002 03:43:35,280 --> 03:43:37,840 PURIFICATION METHOD TO IDENTIFY 5003 03:43:37,840 --> 03:43:39,640 N LINK GLYCOPROTEINS FROM CELL 5004 03:43:39,640 --> 03:43:41,720 LINES AND TWO PROTEINS 5005 03:43:41,720 --> 03:43:43,800 IDENTIFIED WERE CCAM 5 AND LAMP 5006 03:43:43,800 --> 03:43:46,600 1 SO WE DECIDED TO TEST TO SEE 5007 03:43:46,600 --> 03:43:49,880 IF WE CAN OBSERVE THAT CROSS 5008 03:43:49,880 --> 03:43:52,280 LINKING OF GALECTIN 1 TO TWO 5009 03:43:52,280 --> 03:43:55,280 PROTEINS IN OUR CELLS, THIS 5010 03:43:55,280 --> 03:43:58,440 INVOLVES ENINCORPORATING THE 5011 03:43:58,440 --> 03:44:00,840 GLCNGDAZ MEDIATED TO CROSS LINK 5012 03:44:00,840 --> 03:44:04,440 AND ACROSS PROTEIN OF INTEREST 5013 03:44:04,440 --> 03:44:06,880 CCAN 5 WITH LAMP 1 AND GALECTIN 5014 03:44:06,880 --> 03:44:09,200 1 TO LOOK FOR COMPLEX IN THE 5015 03:44:09,200 --> 03:44:11,240 GEL. YOU CAN SEE THE GEL ON THE 5016 03:44:11,240 --> 03:44:12,280 LEFT WE CAN OBSERVE CROSS 5017 03:44:12,280 --> 03:44:16,600 LINKING OF CCAM 5 TO GALECTIN 1 5018 03:44:16,600 --> 03:44:18,040 BOTH IN INTACT CELLS BUT ALSO 5019 03:44:18,040 --> 03:44:21,640 WHEN THE CROSS LINKING WAS 5020 03:44:21,640 --> 03:44:23,680 PERFORMED AFTER WE FIRST LYSED 5021 03:44:23,680 --> 03:44:24,880 THE CELLS, AND PERFORMED CROSS 5022 03:44:24,880 --> 03:44:27,280 LINKING IT WAS THE SAME NO 5023 03:44:27,280 --> 03:44:28,280 MATTER EXPERIMENT. BUT DOING 5024 03:44:28,280 --> 03:44:29,960 WITH LAMP 1 WE DID NOT OBSERVE 5025 03:44:29,960 --> 03:44:31,560 CROSS LINKING WITH INTACT CELLS 5026 03:44:31,560 --> 03:44:34,280 BUT ONLY WHEN WE FIRST LYSE THE 5027 03:44:34,280 --> 03:44:35,560 CELLS AND DID CROSS LINKING. 5028 03:44:35,560 --> 03:44:37,640 THAT WAS INTERESTING BECAUSE 5029 03:44:37,640 --> 03:44:39,880 LAMP 1 DIDN'T SHOW IN PROTEOMICS 5030 03:44:39,880 --> 03:44:42,280 ANALYSIS WHEN GALECTIN WAS A 5031 03:44:42,280 --> 03:44:43,600 APPLIED EXTRA CELLULARLY SO 5032 03:44:43,600 --> 03:44:44,640 SHOWS THE BENEFIT OF CROSS 5033 03:44:44,640 --> 03:44:45,440 LINKING METHOD IS TELLS YOU 5034 03:44:45,440 --> 03:44:46,760 ABOUT THE INTERACTIONS THAT ARE 5035 03:44:46,760 --> 03:44:49,080 OCCURRING IN LIVING CELLS, AND 5036 03:44:49,080 --> 03:44:50,640 WHEREAS AFFINITY PURIFICATION 5037 03:44:50,640 --> 03:44:52,920 SCRAMBLES THE INTERACTIONS SO 5038 03:44:52,920 --> 03:44:54,480 YOU MAY INTRODUCE SPURIOUS 5039 03:44:54,480 --> 03:44:55,160 FINDING INTERACTIONS IN THAT 5040 03:44:55,160 --> 03:44:59,040 WAY. SO WITH THAT OBSERVATION WE 5041 03:44:59,040 --> 03:45:00,520 WENT TO LOOK AT THE CROSS 5042 03:45:00,520 --> 03:45:03,080 LINKING OF ENDOGENOUS GALECTIN 1 5043 03:45:03,080 --> 03:45:05,240 INSIDE THESE CELLS AND WHAT IS 5044 03:45:05,240 --> 03:45:06,760 INTERESTING IS IF WE DON'T TREAT 5045 03:45:06,760 --> 03:45:09,120 THE CELLS WE OBSERVE NO CROSS 5046 03:45:09,120 --> 03:45:12,560 LINKING OF GALECTIN 1 ENDOGENOUS 5047 03:45:12,560 --> 03:45:13,880 GALECTIN 1 INSIDE CELLS BUT IF 5048 03:45:13,880 --> 03:45:17,440 WE TREAT WITH A LYSOSOMAL DAMAGE 5049 03:45:17,440 --> 03:45:19,240 REAGENT, WHICH EXPOSES THE 5050 03:45:19,240 --> 03:45:21,560 CONTENT OF THE LYSOSOME WHICH 5051 03:45:21,560 --> 03:45:23,960 INCLUDES N LINK GLYCOPROTEINS TO 5052 03:45:23,960 --> 03:45:25,440 GALECTINS WHICH ARE IN CYTOPLASM 5053 03:45:25,440 --> 03:45:27,640 WE OBSERVE CROSS LINKING OF 5054 03:45:27,640 --> 03:45:31,040 GALECTIN 1 TO PROTEINS WE ASSUME 5055 03:45:31,040 --> 03:45:33,640 ARE LYSESOMAL PROTEINS. THAT IS 5056 03:45:33,640 --> 03:45:35,880 INHIBITED WE USE INHIBITOR OF 5057 03:45:35,880 --> 03:45:37,600 LYSOSOMAL DAMAGE PATHWAY. SO WE 5058 03:45:37,600 --> 03:45:38,840 ARE CONTINUING TO FOLLOW-UP ON 5059 03:45:38,840 --> 03:45:41,680 THIS TO UNDERSTAND EXACTLY HOW 5060 03:45:41,680 --> 03:45:43,200 GALECTINS INTERACT WITH 5061 03:45:43,200 --> 03:45:44,160 LYSOSOMAL PROTEINS AND 5062 03:45:44,160 --> 03:45:46,840 CONDITIONS OF LYSOSOMAL DAMAGE. 5063 03:45:46,840 --> 03:45:48,920 SO JUST TO CONCLUDE THIS PART 5064 03:45:48,920 --> 03:45:52,240 ABOUT INCORPORATION OF GLCNDAZ 5065 03:45:52,240 --> 03:45:55,560 TO N LINKED PROTEINS, IT WAS 5066 03:45:55,560 --> 03:45:57,240 METABOLICALLY INCORPORATED TO N 5067 03:45:57,240 --> 03:45:58,880 LINK GLYCANS. WE CAN LEARN 5068 03:45:58,880 --> 03:46:00,120 ABOUT THE AMOUNT OF THESE 5069 03:46:00,120 --> 03:46:00,920 BINDING EVENTS THROUGH THE 5070 03:46:00,920 --> 03:46:02,800 INTENSITY OF THE CROSS LINKING, 5071 03:46:02,800 --> 03:46:04,760 WE CAN ISOLATE CROSS LINK 5072 03:46:04,760 --> 03:46:06,360 COMPLEXES AND MOST IMPORTANTLY 5073 03:46:06,360 --> 03:46:08,840 GIVES US CELLS CONTEXT SPECIFIC 5074 03:46:08,840 --> 03:46:10,280 INFORMATION ABOUT RECOGNITION 5075 03:46:10,280 --> 03:46:12,200 EVENTS. WORKING WITH (INAUDIBLE) 5076 03:46:12,200 --> 03:46:13,320 GROUP WE ARE CONTINUING TO 5077 03:46:13,320 --> 03:46:16,520 EVALUATE INCORPORATION OF 5078 03:46:16,520 --> 03:46:19,440 GLCNDAZ INTO OTHER CLASSES OF 5079 03:46:19,440 --> 03:46:20,240 GLYCOCONJUGATES. I WANT TO TELL 5080 03:46:20,240 --> 03:46:21,440 YOU ABOUT THE LAST AIM WHICH IS 5081 03:46:21,440 --> 03:46:25,320 STILL UNDERWAY, AND THIS WAS -- 5082 03:46:25,320 --> 03:46:28,840 WE OUR AIM WAS TO CHARACTERIZE 5083 03:46:28,840 --> 03:46:30,760 IN MORE DETAIL SITES WHICH THE 5084 03:46:30,760 --> 03:46:33,440 CROSS LINKING EVENTS OCCUR AND 5085 03:46:33,440 --> 03:46:36,000 THIS WAS WE DECIDED TO GO ABOUT 5086 03:46:36,000 --> 03:46:39,200 THIS BY LOOKING AT A MODEL 5087 03:46:39,200 --> 03:46:43,040 SYSTEM AND RECOGNITION OF 5088 03:46:43,040 --> 03:46:44,320 O-GlcNac PEPTIDE BY 5089 03:46:44,320 --> 03:46:48,000 PROTEIN 14-3-3 GAMMA, MY LAB 5090 03:46:48,000 --> 03:46:49,440 IDENTIFIED THE INTERACTION AND 5091 03:46:49,440 --> 03:46:50,840 SOLVED THE CRYSTAL STRUCTURE OF 5092 03:46:50,840 --> 03:46:52,040 BINDING INTERACTION A FEW YEARS 5093 03:46:52,040 --> 03:46:55,440 AGO SO WE KNEW WHERE THE GLCNAC 5094 03:46:55,440 --> 03:46:59,760 CONTACT THE 14, 3, 3 PROTEIN. WE 5095 03:46:59,760 --> 03:47:05,360 PREPARED A POPE TIDE WITH 5096 03:47:05,360 --> 03:47:06,640 OGLCNDAZ ON IT AND CROSS LINKED 5097 03:47:06,640 --> 03:47:08,200 THE COMPLEX AND TRYPSIN DIGEST 5098 03:47:08,200 --> 03:47:11,760 IT AND USED THE BIONIC SOFTWARE 5099 03:47:11,760 --> 03:47:13,840 ALSO SUPPORTED THROUGH THE 5100 03:47:13,840 --> 03:47:16,320 COMMON FUND AND SHARED WITH US 5101 03:47:16,320 --> 03:47:20,200 THROUGH THAT EFFORT TO LOOK FOR 5102 03:47:20,200 --> 03:47:21,040 ADDUCTS THAT REPRESENT CROSSING 5103 03:47:21,040 --> 03:47:24,760 OF PEPTIDE TO 14-3-3 PROTEIN. 5104 03:47:24,760 --> 03:47:26,320 WESTERN ABLE TO FIND ONE SPECIES 5105 03:47:26,320 --> 03:47:28,200 THAT CORRESPONDED TO THIS, WE 5106 03:47:28,200 --> 03:47:30,600 ARE LOOKING TO LOOK AT OTHER 5107 03:47:30,600 --> 03:47:33,040 MODEL SYSTEMS TO SEE IF WE CAN 5108 03:47:33,040 --> 03:47:34,480 SEE THIS KIND OF CROSS LINKING 5109 03:47:34,480 --> 03:47:36,120 EVENTS IN SYSTEMS WHERE WE DON'T 5110 03:47:36,120 --> 03:47:37,600 HAVE STRUCTURAL INFORMATION. SO 5111 03:47:37,600 --> 03:47:39,840 THAT WORK IS UNDERWAY. ALL 5112 03:47:39,840 --> 03:47:47,360 RIGHT. SO THIS IS MY SUMMARY OF 5113 03:47:47,360 --> 03:47:49,640 OUR ENTIRE PROJECT WE PUT 5114 03:47:49,640 --> 03:47:51,520 TOGETHER. THE CONRAD LAB CREATE 5115 03:47:51,520 --> 03:47:53,000 AD FLUORESCE SENT REPORTER OF 5116 03:47:53,000 --> 03:47:54,080 O-GlcNac LEVELS BASED ON 5117 03:47:54,080 --> 03:47:56,080 THE SPLICING OF OGT TRANSCRIPT. 5118 03:47:56,080 --> 03:47:57,760 WE DEVELOPED A MORE RELIABLE 5119 03:47:57,760 --> 03:48:01,520 SYNTHETIC ROUTE TO MAKE HEXOSE 5120 03:48:01,520 --> 03:48:04,800 MIEN 1 PHOSPHATE ANALOGS USING 5121 03:48:04,800 --> 03:48:06,760 FOR OTHER COMPOUNDS TO FIND 5122 03:48:06,760 --> 03:48:08,040 SUGARS WE WERE INTERESTED IN AND 5123 03:48:08,040 --> 03:48:11,120 WE DEMONSTRATED THE GLCNDAZ 5124 03:48:11,120 --> 03:48:12,920 COMPOUND IS INCORPORATED TO N 5125 03:48:12,920 --> 03:48:15,000 LINK GLYCANS AND USEFUL TO PROBE 5126 03:48:15,000 --> 03:48:16,120 INTERACTIONS. WE ARE CONTINUING 5127 03:48:16,120 --> 03:48:17,800 TO INCORPORATE EVALUATE 5128 03:48:17,800 --> 03:48:20,200 INCORPORATION OF THESE UNNATURAL 5129 03:48:20,200 --> 03:48:22,720 SUGARS AND WRAPPING UP THAT MASS 5130 03:48:22,720 --> 03:48:23,800 SPEC ANALYSIS OF THE SITE TO 5131 03:48:23,800 --> 03:48:26,240 CROSS LINKING. SO I WANT TO WRAP 5132 03:48:26,240 --> 03:48:28,960 UP WITH A COUPLE OF WORDS ABOUT 5133 03:48:28,960 --> 03:48:30,800 DISSEMINATION. WE WERE THROUGH 5134 03:48:30,800 --> 03:48:32,120 THE EFFORTS OF CATHERINE WE WERE 5135 03:48:32,120 --> 03:48:33,400 ABLE TO PUT TOGETHER THIS 5136 03:48:33,400 --> 03:48:35,200 CURRENT PROTOCOLS TO SHARE IN 5137 03:48:35,200 --> 03:48:38,480 CURRENT PROTOCOLS. OUR REAGENT 5138 03:48:38,480 --> 03:48:41,440 TO INTRODUCE UNNATURAL UAP IS 5139 03:48:41,440 --> 03:48:42,800 AVAILABLE FROM ADD GENE AND YOU 5140 03:48:42,800 --> 03:48:46,760 HEARD THERE IS REAGENTS FROM 5141 03:48:46,760 --> 03:48:48,240 NATASHA'S LAB AS WELL. FOR NOW 5142 03:48:48,240 --> 03:48:51,480 YOU CAN REQUEST COMPOUND FROM 5143 03:48:51,480 --> 03:48:53,200 US, BUT CHRIS IS WORKING ON 5144 03:48:53,200 --> 03:48:55,040 PREPARING FOR COMMERCIAL 5145 03:48:55,040 --> 03:48:57,520 AVAILABILITY. TOCRIS. THESE ARE 5146 03:48:57,520 --> 03:48:58,920 SOME OF THE PUBLICATIONS WE HAVE 5147 03:48:58,920 --> 03:49:00,400 HAD AND I WOULD LIKE TO THANK 5148 03:49:00,400 --> 03:49:02,840 ALL MY COLLABORATORS BOTH AT UT 5149 03:49:02,840 --> 03:49:03,840 SOUTH WESTERN AND AT UNIVERSITY 5150 03:49:03,840 --> 03:49:05,960 OF GEORGIA, IT'S BEEN GREAT TO 5151 03:49:05,960 --> 03:49:07,600 WORK ON THIS PROJECT WITH ALL OF 5152 03:49:07,600 --> 03:49:08,400 THEM. THANK YOU FOR YOUR 5153 03:49:08,400 --> 03:49:09,360 ATTENTION. 5154 03:49:09,360 --> 03:49:15,920 [APPLAUSE] 5155 03:49:15,920 --> 03:49:22,160 >> OUR NEXT SPEAKER IS DR. 5156 03:49:22,160 --> 03:49:31,400 CHRISTINE WOO, HARVARD. AS HAS 5157 03:49:31,400 --> 03:49:33,600 BEEN BEAUTIFULLY INTRODUCED BY 5158 03:49:33,600 --> 03:49:35,360 NATASHA NOW JEN THIS 5159 03:49:35,360 --> 03:49:37,520 CARBOHYDRATE HERE HOLDS 5160 03:49:37,520 --> 03:49:38,480 O-GlcNac, IS FOUND ON 5161 03:49:38,480 --> 03:49:40,360 SERINE 3 ANINE RESIDUES WITHIN 5162 03:49:40,360 --> 03:49:43,160 THE CELL NUCLEAR CYTOPLASMIC AND 5163 03:49:43,160 --> 03:49:44,520 MITOCHONDRIAL PROTEINS. IT IS A 5164 03:49:44,520 --> 03:49:48,760 METABOLIC SENSOR THAT IS QUITE 5165 03:49:48,760 --> 03:49:50,080 IMPORTANT AND I WON'T REITERATE 5166 03:49:50,080 --> 03:49:52,400 ALL THE DETAILS OF WHY YOU 5167 03:49:52,400 --> 03:49:54,440 SHOULD THINK ABOUT THIS SUGAR 5168 03:49:54,440 --> 03:49:56,480 MOLECULE, BUT THE KEY PART THAT 5169 03:49:56,480 --> 03:50:01,480 FASCINATED US IS THAT THIS SMALL 5170 03:50:01,480 --> 03:50:04,080 MOLECULE IS GLYCOSYLATION EVENT 5171 03:50:04,080 --> 03:50:05,960 OCCURS ON ALL THESE PROTEINS 5172 03:50:05,960 --> 03:50:08,160 SUBSTRATES WITH THE ACTIVITY OF 5173 03:50:08,160 --> 03:50:13,360 A SINGLE SET OF ENZYMES, WRITER 5174 03:50:13,360 --> 03:50:15,280 O-GlcNac TRANSFER RACE OR 5175 03:50:15,280 --> 03:50:19,400 OGT AND ONE ERASER OGA. THERE'S 5176 03:50:19,400 --> 03:50:20,680 DYNAMICS WEAN THE TWO ENZYMES 5177 03:50:20,680 --> 03:50:22,440 THAT IS REGULATING THE LEVELS OF 5178 03:50:22,440 --> 03:50:26,680 THIS SUGAR. SO THESE TWO ENZYMES 5179 03:50:26,680 --> 03:50:28,720 ARE QUITE ESSENTIAL, THEY ARE 5180 03:50:28,720 --> 03:50:30,440 REQUIRED FOR CELLULAR DIVISION 5181 03:50:30,440 --> 03:50:33,360 AND CELLULAR PROLIFERATION. THEY 5182 03:50:33,360 --> 03:50:34,280 ALSO STABILIZE NUMBER OF 5183 03:50:34,280 --> 03:50:36,040 PROTEINS IN OUR CELLS INCLUDING 5184 03:50:36,040 --> 03:50:39,920 ALPHA SYNUCLEIN, WHICH WHEN 5185 03:50:39,920 --> 03:50:42,360 ALPHA SYNUCLEIN IS GLYCOSYLATED 5186 03:50:42,360 --> 03:50:44,880 IT IS SOLUBLE BUT WHEN 5187 03:50:44,880 --> 03:50:45,880 DEGLYCOSYLATED IT FORMS 5188 03:50:45,880 --> 03:50:47,480 AGGREGATES AND AFTER LUNCH AS WE 5189 03:50:47,480 --> 03:50:50,640 ALL FEEL SATIATED, THAT IS 5190 03:50:50,640 --> 03:50:51,360 PLAYING O-GlcNac IS 5191 03:50:51,360 --> 03:50:52,840 PLAYING A ROLE IN OUR BRAINS AND 5192 03:50:52,840 --> 03:50:55,360 TELLING US THAT WE HAVE ENOUGH 5193 03:50:55,360 --> 03:50:59,000 SUGAR AND ENERGY AND IF WE LOSE 5194 03:50:59,000 --> 03:51:00,160 OGT IN CERTAIN PARTS OF THE 5195 03:51:00,160 --> 03:51:07,000 BRAIN THIS CAN INDUCE DIABETIC 5196 03:51:07,000 --> 03:51:08,760 PHENOTYPE AND NOT BEING ABLE TO 5197 03:51:08,760 --> 03:51:09,760 SIGNAL WHEN HOW MUCH FOOD IS 5198 03:51:09,760 --> 03:51:12,200 AVAILABLE. SO WHAT IS QUITE 5199 03:51:12,200 --> 03:51:15,120 INTERESTING TO US AND I THINK 5200 03:51:15,120 --> 03:51:17,320 REALLY WITH INNOVATIONS WITH 5201 03:51:17,320 --> 03:51:18,480 ASSISTANCE OF INNOVATIONS ON 5202 03:51:18,480 --> 03:51:20,000 MASS SPEC THAT WE WILL HEAR 5203 03:51:20,000 --> 03:51:23,600 ABOUT IN THE COMING DAY OR TWO, 5204 03:51:23,600 --> 03:51:24,920 HAS BEEN NOW THAT WE HAVE THESE 5205 03:51:24,920 --> 03:51:27,680 DEEPER DEEPER MAPS OF WHERE AND 5206 03:51:27,680 --> 03:51:29,120 WHEN O-GlcNac IS OCCURRING 5207 03:51:29,120 --> 03:51:31,320 BY MASS SPEC WE NOW KNOW THIS 5208 03:51:31,320 --> 03:51:33,560 SUGAR MOLECULE IS FOUND ON OVER 5209 03:51:33,560 --> 03:51:35,880 5,000 DIFFERENT PROTEINS. MANY 5210 03:51:35,880 --> 03:51:37,200 OF WHICH SPAN NUMBER OF 5211 03:51:37,200 --> 03:51:39,720 SIGNALING PROTEINS LIKE 5212 03:51:39,720 --> 03:51:40,560 TRANSCRIPTION FACTORS, RNA 5213 03:51:40,560 --> 03:51:43,440 BINDING PROTEINS, KINASE AND E 3 5214 03:51:43,440 --> 03:51:44,880 LIGASE. SO CENTRAL QUESTION WE 5215 03:51:44,880 --> 03:51:47,320 STARTED TO ASK IN MY LAB IS HOW 5216 03:51:47,320 --> 03:51:48,760 DOES O-GlcNac TAYLOR 5217 03:51:48,760 --> 03:51:51,560 PROTEIN FUNCTIONS IN CELLS? 5218 03:51:51,560 --> 03:51:52,880 SECONDARILY WE CAN ANSWER THE 5219 03:51:52,880 --> 03:51:54,920 QUESTION HOW AND WHERE TO USE 5220 03:51:54,920 --> 03:51:56,040 O-GlcNac TO MITIGATE HUMAN 5221 03:51:56,040 --> 03:51:58,040 DISEASES. SO AS WE CONTEMPLATED 5222 03:51:58,040 --> 03:52:00,000 THIS QUESTION AND HOW WE CAN 5223 03:52:00,000 --> 03:52:02,120 ANSWER IT, WE REALIZE WE NEED AD 5224 03:52:02,120 --> 03:52:03,680 BETTER WAY TO ACTUALLY 5225 03:52:03,680 --> 03:52:05,400 MANIPULATE O-GlcNac IN A 5226 03:52:05,400 --> 03:52:08,000 CELLULAR CONTEXT ABLE TO ASK HOW 5227 03:52:08,000 --> 03:52:08,520 O-GlcNac FUNCTIONS AND 5228 03:52:08,520 --> 03:52:11,040 THEN HOW WE CAN USE IT FOR 5229 03:52:11,040 --> 03:52:13,760 BENEFICIAL INTERCEPTION. AS WE 5230 03:52:13,760 --> 03:52:14,720 THOUGHT ABOUT DIFFERENT OPTIONS 5231 03:52:14,720 --> 03:52:16,600 FOR USING MANIPULATING 5232 03:52:16,600 --> 03:52:18,080 O-GlcNac, WE CAME UP WITH 5233 03:52:18,080 --> 03:52:20,120 THIS IDEA WE THOUGHT WELL, IF 5234 03:52:20,120 --> 03:52:21,200 ONLY WE COULD TAKE A TARGET 5235 03:52:21,200 --> 03:52:23,000 PROTEIN AND RECRUIT 5236 03:52:23,000 --> 03:52:25,880 O-GlcNac TO IT AND IN THIS 5237 03:52:25,880 --> 03:52:28,040 WAY BY RECRUITING THAT 5238 03:52:28,040 --> 03:52:30,200 PROXIMITY, THAT O-GlcNac 5239 03:52:30,200 --> 03:52:32,760 TRANSFERASE THROUGH PROXIMITY 5240 03:52:32,760 --> 03:52:34,320 ENHANCING 5241 03:52:34,320 --> 03:52:35,520 O-GglcNaCYLATION ON THAT 5242 03:52:35,520 --> 03:52:38,360 PROTEIN. CONTRAST IF WE REDUCE 5243 03:52:38,360 --> 03:52:42,320 THE SINGER E RAZOR 5244 03:52:42,320 --> 03:52:43,160 O-GlcNacASE WE CAN 5245 03:52:43,160 --> 03:52:44,600 DEGLYCOSYLATE IT. AND IF WE CAN 5246 03:52:44,600 --> 03:52:47,520 CYCLE SELECTIVELY ON TARGET 5247 03:52:47,520 --> 03:52:50,400 PROTEIN IN CELLULAR CONTEXT WE 5248 03:52:50,400 --> 03:52:53,320 CAN CHARACTERIZE RULE OF THAT 5249 03:52:53,320 --> 03:52:55,120 GLYCOSYLATION EVENT EVENT. AS WE 5250 03:52:55,120 --> 03:52:56,960 THOUGHT ABOUT WAYS TO RECRUIT 5251 03:52:56,960 --> 03:52:59,960 AND RECOGNIZE TARGET PROTEINS, 5252 03:52:59,960 --> 03:53:01,360 IT BECAME -- OBVIOUSLY CELLS 5253 03:53:01,360 --> 03:53:02,880 KNOW HOW TO DO THIS AND THEY 5254 03:53:02,880 --> 03:53:04,760 KNOW HOW TO DO IT USING ANTIBODY 5255 03:53:04,760 --> 03:53:06,760 SO ANTIBODIES ARE HIGHLY 5256 03:53:06,760 --> 03:53:09,200 SPECIALIZED PROTEIN RECOGNITION 5257 03:53:09,200 --> 03:53:10,640 SYSTEMS BUT PRIMARILY FOUND ON 5258 03:53:10,640 --> 03:53:12,760 THE OUTSIDE OF CELLS, SO 5259 03:53:12,760 --> 03:53:13,920 ANTIBODIES RECOGNIZE NUMBER OF 5260 03:53:13,920 --> 03:53:15,560 PROTEINS WITH REALLY HIGH 5261 03:53:15,560 --> 03:53:16,960 SELECTIVITY AND SENSITIVITY ON 5262 03:53:16,960 --> 03:53:19,320 CELL SURFACE. THIS DOESN'T BODE 5263 03:53:19,320 --> 03:53:21,440 WELL FOR INTRACELLULAR PROTEINS, 5264 03:53:21,440 --> 03:53:22,920 BUT FORTUNATELY THERE IS 5265 03:53:22,920 --> 03:53:26,200 EMERGING RESEARCH IN THE AREA OF 5266 03:53:26,200 --> 03:53:31,120 NANOBODIES AND OTHER SINGLE 5267 03:53:31,120 --> 03:53:33,360 DOMAIN PROTEIN BINDERS, WHICH WE 5268 03:53:33,360 --> 03:53:35,600 ARE PARTICULARLY ATTRACTED TO 5269 03:53:35,600 --> 03:53:37,280 NANOBODIES, THEY ARE ESSENTIALLY 5270 03:53:37,280 --> 03:53:39,200 SMALL ANTIBODIES THAT 5271 03:53:39,200 --> 03:53:41,160 SPECIFICALLY DERIVE FROM CAMELS. 5272 03:53:41,160 --> 03:53:44,760 AND THESE SMALL ANTIBODIES ARE 5273 03:53:44,760 --> 03:53:46,680 ESSENTIALLY HIGH AFFINITY 5274 03:53:46,680 --> 03:53:47,960 PROTEIN RECOGNITION SYSTEMS THAT 5275 03:53:47,960 --> 03:53:50,920 ARE NOW ALSO SMALL SINGLE DOMAIN 5276 03:53:50,920 --> 03:53:53,520 PROTEIN BINDERS THAT WE CAN FUSE 5277 03:53:53,520 --> 03:53:58,760 TO THE O-GlcNac TRANSFER 5278 03:53:58,760 --> 03:53:59,320 TRANSFERRIST AND 5279 03:53:59,320 --> 03:54:00,080 O-GlcNacASE. SO I WILL 5280 03:54:00,080 --> 03:54:01,840 TELL YOU ABOUT THE DESIGN OF THE 5281 03:54:01,840 --> 03:54:04,360 SYSTEM AND HOW WE USE IT TO DATE 5282 03:54:04,360 --> 03:54:06,360 AND DISSEMINATION AND FUTURE 5283 03:54:06,360 --> 03:54:09,520 PLANS. SO IN TERMS OF DESIGNING 5284 03:54:09,520 --> 03:54:11,720 THE WRITER TARGET PROTEIN 5285 03:54:11,720 --> 03:54:12,960 WRITER, OF O-GlcNac, WE 5286 03:54:12,960 --> 03:54:14,840 FIRST LOOK AT STRUCTURE OF 5287 03:54:14,840 --> 03:54:15,360 O-GlcNac TRANSFERASE WE 5288 03:54:15,360 --> 03:54:17,560 ARE SHOWING YOU LINEAR STRUCTURE 5289 03:54:17,560 --> 03:54:19,080 AS AS WELL AS CRYSTAL STRUCTURE 5290 03:54:19,080 --> 03:54:22,120 DOWN HERE. SO O-GlcNac 5291 03:54:22,120 --> 03:54:23,520 TRANSFERASE HAS TWO PRIMARY 5292 03:54:23,520 --> 03:54:25,440 DOMAINS, ONE IS CATALYTIC DOMAIN 5293 03:54:25,440 --> 03:54:26,560 SHOWN HERE IN YELLOW AND THE 5294 03:54:26,560 --> 03:54:28,560 OTHER PART IS THIS HETERO 5295 03:54:28,560 --> 03:54:32,000 PEPTIDE REPEAT DOMAIN IN PURPLE. 5296 03:54:32,000 --> 03:54:33,440 THE CATALYTIC DOMAIN WHERE ALL 5297 03:54:33,440 --> 03:54:36,160 THE ACTION IS HAPPENING FOR THE 5298 03:54:36,160 --> 03:54:37,840 GLYCOSYLATION EVENT IS 5299 03:54:37,840 --> 03:54:39,400 HIGHLIGHTED HERE IN SPECIFIC 5300 03:54:39,400 --> 03:54:41,920 AMINO ACID RESIDUES IN RED. ALL 5301 03:54:41,920 --> 03:54:45,560 THAT ACTION IS SHOWN HERE BUT 5302 03:54:45,560 --> 03:54:46,360 HOW O-GlcNac RELATION 5303 03:54:46,360 --> 03:54:48,440 EVENTS OCCUR IS POSITIONED ALL 5304 03:54:48,440 --> 03:54:51,880 BY THE TETRA TRICO PEPTIDE 5305 03:54:51,880 --> 03:54:53,160 REPEAT DOMAIN AND CONTACTS 5306 03:54:53,160 --> 03:54:54,520 THROUGHOUT THIS DOMAIN. WHAT WE 5307 03:54:54,520 --> 03:54:56,080 ENVISION IS PERHAPS REDUCE AND 5308 03:54:56,080 --> 03:54:58,520 REMOVE THIS DOMAIN AND REP PLACE 5309 03:54:58,520 --> 03:55:01,520 WITH A NANOBODY AND HOW 5310 03:55:01,520 --> 03:55:04,080 O-GlcNac TRANSFER AIS IS 5311 03:55:04,080 --> 03:55:05,960 DOING PROXIMITY INDUCTION IN 5312 03:55:05,960 --> 03:55:07,440 DOMAIN AND REPLACING SUBSTITUTE 5313 03:55:07,440 --> 03:55:08,720 WITH ANTIBODY. SO THAT IS WHAT 5314 03:55:08,720 --> 03:55:11,600 WE DID. WE TOOK THAT TPR DOMAIN 5315 03:55:11,600 --> 03:55:13,040 AND TURNS OUT YOU CAN'T GET RID 5316 03:55:13,040 --> 03:55:15,480 OF IT COMPLETELY, YOU CAN GET IT 5317 03:55:15,480 --> 03:55:18,520 DOWN TO ABOUT FOUR TETRATRICO 5318 03:55:18,520 --> 03:55:22,640 PEPTIDE REPIECE AND GET MOST 5319 03:55:22,640 --> 03:55:24,040 CONSENSUS SEQUENCE BUT REDUCE 5320 03:55:24,040 --> 03:55:26,160 BACKGROUND AND LIMIT ENZYME 5321 03:55:26,160 --> 03:55:27,560 ABILITY TO EFFECTIVE BROAD SWATH 5322 03:55:27,560 --> 03:55:29,840 OF SUBSTRATES, AND THEN 5323 03:55:29,840 --> 03:55:31,600 SUBSTITUTE THAT WITH NANOO BODY. 5324 03:55:31,600 --> 03:55:33,360 SO AGAIN THE IDEA IS THAT WE 5325 03:55:33,360 --> 03:55:34,720 HAVE THIS TARGET PROTEIN WE ARE 5326 03:55:34,720 --> 03:55:35,880 INTERESTED IN, AND HAPPENS TO 5327 03:55:35,880 --> 03:55:37,480 HAVE A TAG ON IT, WE CAN RECRUIT 5328 03:55:37,480 --> 03:55:39,160 THAT THROUGH THE NANOBODY OR IF 5329 03:55:39,160 --> 03:55:40,920 WE HAPPEN TO HAVE ONE THAT 5330 03:55:40,920 --> 03:55:42,480 RECOGNIZES ENDOGENOUS TARGET 5331 03:55:42,480 --> 03:55:43,920 PROTEIN WE CAN RECRUIT IT TO 5332 03:55:43,920 --> 03:55:45,920 THAT TARGET PROTEIN AS WELL. 5333 03:55:45,920 --> 03:55:48,240 WITHOUT AFFECTING BROADER 5334 03:55:48,240 --> 03:55:50,360 GLYCOPROTEOME. SO SOME OF OUR 5335 03:55:50,360 --> 03:55:52,000 FAVORITE NANOBODIES ARE SHOWN 5336 03:55:52,000 --> 03:55:54,960 HERE INCLUDING NANOBODIES 5337 03:55:54,960 --> 03:55:57,360 AGAINST GFP SEVERAL AFFINITIES, 5338 03:55:57,360 --> 03:55:58,600 NANOBODIES THAT DIRECTLY 5339 03:55:58,600 --> 03:55:59,800 RECOGNIZE PROTEINS LIKE ALPHA 5340 03:55:59,800 --> 03:56:04,200 SYNUCLEIN AND OTHER PEPTIDES 5341 03:56:04,200 --> 03:56:06,760 TABS 6 PEPTIDE OR BC 2 PEPTIDE. 5342 03:56:06,760 --> 03:56:08,240 THIS WORK WAS ESTABLISHED BY ONE 5343 03:56:08,240 --> 03:56:11,240 OF MY FIRST GRAD STUDENTS NOW 5344 03:56:11,240 --> 03:56:15,120 GRADUATED AT STANFORD IN 5345 03:56:15,120 --> 03:56:18,080 POST-DOCTORAL FELLOWSHIP DANIEL 5346 03:56:18,080 --> 03:56:19,800 RAMMIER RUIZ. SO YOU TAKE THE 5347 03:56:19,800 --> 03:56:22,000 NANOBODY AND INTRODUCE TO CELLS 5348 03:56:22,000 --> 03:56:26,760 LINES FOR TRANSFECTION AND 5349 03:56:26,760 --> 03:56:28,560 CHARACTERIZE O-GlcNacATEED 5350 03:56:28,560 --> 03:56:30,240 PROTEOME SO THIS IS A WRITER OF 5351 03:56:30,240 --> 03:56:33,000 O-GlcNac WE NEED AN ERASER 5352 03:56:33,000 --> 03:56:36,200 TO ERASE THE MODIFICATION. NOW 5353 03:56:36,200 --> 03:56:38,240 INSTEAD OF RECRUITING 5354 03:56:38,240 --> 03:56:39,760 O-GlcNac RELATION WE WANT 5355 03:56:39,760 --> 03:56:42,360 TO REMOVE IT. FORTUNATELY AS WE 5356 03:56:42,360 --> 03:56:45,440 STARTED TO THINK ABOUT THIS 5357 03:56:45,440 --> 03:56:46,400 SEVERAL CRYSTAL STRUCTURE OF OGA 5358 03:56:46,400 --> 03:56:49,200 WERE PUBLISHED AND WHEN WE LOOK 5359 03:56:49,200 --> 03:56:52,200 AD IF THE STRUCTURE AND THE 5360 03:56:52,200 --> 03:56:53,760 RELATIONSHIP WORK THEY DID WAS 5361 03:56:53,760 --> 03:56:55,520 REALLY BEAUTIFUL ABOUT THE OGA 5362 03:56:55,520 --> 03:56:57,520 STRUCTURES, ACTUALLY A 5363 03:56:57,520 --> 03:56:58,640 HOMODIMER, WITH ONE EQUIVALENT 5364 03:56:58,640 --> 03:57:01,280 OF THE OGA SHOWN HERE IN DARK 5365 03:57:01,280 --> 03:57:03,960 RED AND DARK BLUE AND ANOTHER IN 5366 03:57:03,960 --> 03:57:05,840 LIGHT BLUE AND LIGHT PINK. 5367 03:57:05,840 --> 03:57:07,160 CORRESPONDING TO THE CATALYTIC 5368 03:57:07,160 --> 03:57:09,280 DOMAIN IN PINK AND THE STOCK 5369 03:57:09,280 --> 03:57:14,080 DOMAIN SHOWN IN BLUE. ONE OF THE 5370 03:57:14,080 --> 03:57:16,040 REALIZATION IS THAT WE HAD FROM 5371 03:57:16,040 --> 03:57:18,280 LOOKING AT THIS STRUCTURE AND 5372 03:57:18,280 --> 03:57:21,080 FROM RECENTLY PUBLISHED WORK WAS 5373 03:57:21,080 --> 03:57:22,440 THAT PERHAPS WE COULD AGAIN 5374 03:57:22,440 --> 03:57:23,920 REMOVE REDUCE SOME OF THE 5375 03:57:23,920 --> 03:57:25,800 ACTIVITY OF THE ENZYME AND 5376 03:57:25,800 --> 03:57:26,960 REPLACE WITH NANOBODY, IN THIS 5377 03:57:26,960 --> 03:57:29,600 CASE WE CAPITALIZE ON A SPECIFIC 5378 03:57:29,600 --> 03:57:32,560 NATURAL CLEAVAGE EVENT FOUND 5379 03:57:32,560 --> 03:57:34,080 WITHIN OGA SHOWN HERE THAT 5380 03:57:34,080 --> 03:57:35,720 SEPARATES THE CATALYTIC DOMAIN 5381 03:57:35,720 --> 03:57:39,240 FROM STOCK DOMAIN REDUCING THE 5382 03:57:39,240 --> 03:57:41,120 FORMATION OF THIS CALLED 5383 03:57:41,120 --> 03:57:43,960 PLAQUES. UNFORTUNATELY WE CAN 5384 03:57:43,960 --> 03:57:46,320 THEN -- WE USING A SYSTEMATIC 5385 03:57:46,320 --> 03:57:47,760 ENGINEERING APPROACH REALLY 5386 03:57:47,760 --> 03:57:50,920 PIONEERED AND HERALDED BY YUN GE 5387 03:57:50,920 --> 03:57:53,240 FORMER POST-DOC IN MY LAB NOW AT 5388 03:57:53,240 --> 03:57:55,440 BAY LABORATORY IN HER OWN 5389 03:57:55,440 --> 03:57:57,000 INDEPENDENT GROUP REALLY 5390 03:57:57,000 --> 03:57:58,080 SYSTEMATICALLY ENGINEERED A 5391 03:57:58,080 --> 03:58:00,240 SPLIT OGA SYSTEM CONSISTING OF 5392 03:58:00,240 --> 03:58:04,120 THE CATALYTIC DOMAIN WITH THE 5393 03:58:04,120 --> 03:58:05,680 STALK DOMAIN AS INDEPENDENT 5394 03:58:05,680 --> 03:58:07,720 FRAGMENT. THIS SYSTEM ON ITS OWN 5395 03:58:07,720 --> 03:58:09,560 HAS LIMITED CATALYTIC ACTIVITY 5396 03:58:09,560 --> 03:58:11,280 ON ENDOGENOUS SUBSTRATES AND 5397 03:58:11,280 --> 03:58:13,320 ONLY WHEN WE INTRODUCED THE 5398 03:58:13,320 --> 03:58:14,480 NANOBODY TO THAT STALK DOMAIN 5399 03:58:14,480 --> 03:58:21,120 CAN WE RECOVER THE ACTIVITY. SO 5400 03:58:21,120 --> 03:58:23,200 IN AIM ONE WAS FOCUSED ON 5401 03:58:23,200 --> 03:58:24,760 WRITING AND SYSTEMATICALLY 5402 03:58:24,760 --> 03:58:26,400 CHARACTERIZING WITH NANOBODIES 5403 03:58:26,400 --> 03:58:29,280 AND TWO PROPALE WAS LOOKING AT 5404 03:58:29,280 --> 03:58:31,280 ERASING THE MODIFICATION AND NOW 5405 03:58:31,280 --> 03:58:33,160 THAT WE HAVE THESE TOOL SYSTEMS 5406 03:58:33,160 --> 03:58:37,040 WE CAN USE THEM FOR 5407 03:58:37,040 --> 03:58:41,520 CHARACTERIZING 5408 03:58:41,520 --> 03:58:43,400 O-GlcNaCYLATION EVENTS AS 5409 03:58:43,400 --> 03:58:45,560 TOOLS. IF YOU WANT YOUR OWN 5410 03:58:45,560 --> 03:58:47,840 NANOBODY INFUSED ENZYMES FOR 5411 03:58:47,840 --> 03:58:49,520 OTHER GLYCOSYLATION EVENTS WE 5412 03:58:49,520 --> 03:58:51,680 LEARNED THROUGH OUR WORK THAT 5413 03:58:51,680 --> 03:58:54,680 THIS SYSTEMATIC APPROACH FIRST 5414 03:58:54,680 --> 03:58:57,120 REDUCING ACTIVITY OF THE ENZYMES 5415 03:58:57,120 --> 03:58:59,160 ENDOGENOUS SUBSTRATE RECOGNITION 5416 03:58:59,160 --> 03:59:01,120 MECHANISMS, BY WHICH YOU CAN 5417 03:59:01,120 --> 03:59:02,240 LEARN ABOUT THE STRUCTURE 5418 03:59:02,240 --> 03:59:05,360 ACTIVITY RELATIONSHIPS OF THESE 5419 03:59:05,360 --> 03:59:07,560 ENZYME SYSTEMS, TO LIMIT THE 5420 03:59:07,560 --> 03:59:08,760 BACKGROUND AND FOLLOWED BY 5421 03:59:08,760 --> 03:59:10,760 RECOVERING THE ACTIVITY WITH 5422 03:59:10,760 --> 03:59:11,720 NANOBODY CAN GIVE YOU SELECTIVE 5423 03:59:11,720 --> 03:59:17,520 SYSTEMS. IN THE LAST FEW MINUTES 5424 03:59:17,520 --> 03:59:20,280 I WANT TO SHARE A KEY QUESTION I 5425 03:59:20,280 --> 03:59:21,960 THINK THAT MAJOR MOTIVATION OF 5426 03:59:21,960 --> 03:59:24,080 THE COMMON FUND AND OURS AS WELL 5427 03:59:24,080 --> 03:59:25,200 IS HOW I CAN USE THE 5428 03:59:25,200 --> 03:59:26,440 O-GlcNac WRITER AND ERASER 5429 03:59:26,440 --> 03:59:27,800 ON MY TARGET PROTEIN. SO I 5430 03:59:27,800 --> 03:59:30,120 WANTED TO TELL ONE REALLY BRIEF 5431 03:59:30,120 --> 03:59:34,200 VIGNETTE PIONEERED HERE BY PAUL 5432 03:59:34,200 --> 03:59:36,440 STEIN ANOTHER SOON TO BE 5433 03:59:36,440 --> 03:59:40,120 GRADUATED NOW DR. PAUL SCHWEIN 5434 03:59:40,120 --> 03:59:41,160 STUDENT MANY MY LAB. PAUL IS 5435 03:59:41,160 --> 03:59:42,640 INTERESTED IN CHARACTERIZING THE 5436 03:59:42,640 --> 03:59:45,520 INTERPLAY WITH O-GlcNac 5437 03:59:45,520 --> 03:59:47,960 AND PHOSPHORYLATION SPECIFICALLY 5438 03:59:47,960 --> 03:59:48,600 THROUGH HOW O-GlcNac CAN 5439 03:59:48,600 --> 03:59:53,160 EFFECT KINASES DIRECTLY ON THE 5440 03:59:53,160 --> 03:59:54,040 ENZYMES THAT WRITE 5441 03:59:54,040 --> 03:59:55,560 PHOSPHORYLATION. SO HE PICKED UP 5442 03:59:55,560 --> 04:00:00,280 ON PRECEDENT OUT OF THE COLE LAB 5443 04:00:00,280 --> 04:00:05,160 WHERE WHO HAD SHOWN THAT CASEIN 5444 04:00:05,160 --> 04:00:07,280 KINASE 2 UBIQUITOUSLY EXPRESSED 5445 04:00:07,280 --> 04:00:09,280 ONE OF THE FIRST KINASES 5446 04:00:09,280 --> 04:00:10,640 DISCOVERED, UBIQUITOUSLY 5447 04:00:10,640 --> 04:00:11,680 EXPRESSED AND ACTIVE WITH 5448 04:00:11,680 --> 04:00:14,080 HUNDREDS OF DIFFERENT 5449 04:00:14,080 --> 04:00:19,360 SUBSTRATES. THIS KINASE HAS A 5450 04:00:19,360 --> 04:00:21,360 SINGLE GLYCOCYTE THAT IN VITRO 5451 04:00:21,360 --> 04:00:22,840 SHOWS PROTEIN PROTEIN 5452 04:00:22,840 --> 04:00:24,040 INTERACTIONS AS WELL AS 5453 04:00:24,040 --> 04:00:25,720 SUBSTRATES AND PAUL WANTED TO 5454 04:00:25,720 --> 04:00:27,320 USE THE TOOLS WE DEVELOPED HERE 5455 04:00:27,320 --> 04:00:30,600 AS PARTS OF THE COMMON FUND TO 5456 04:00:30,600 --> 04:00:33,360 CHARACTERIZE WHETHER OR NOT THIS 5457 04:00:33,360 --> 04:00:36,360 SINGLE S 347 GLYCOSIDE ALTERS CK 5458 04:00:36,360 --> 04:00:38,480 2 SUBSTRATES. SO THE IDEA TO 5459 04:00:38,480 --> 04:00:40,240 USE THESE WRITERS AND ERASERS IS 5460 04:00:40,240 --> 04:00:42,840 YOU CAN APPLY THE NANOBODY OGT 5461 04:00:42,840 --> 04:00:44,960 SYSTEM TO INTRODUCE THE 5462 04:00:44,960 --> 04:00:45,960 GLYCOSYLATION EVENT, HERE 5463 04:00:45,960 --> 04:00:49,880 THROUGH A GFP TAGGED CP 2 ALPHA. 5464 04:00:49,880 --> 04:00:52,560 AND YOU CAN APPLY NANOBODY OGA 5465 04:00:52,560 --> 04:00:56,840 SYSTEM TO REDUCE IT. THE 5466 04:00:56,840 --> 04:00:58,080 IMPORTANTLY FOR EVERY ACTIVE 5467 04:00:58,080 --> 04:01:00,360 ENZYME WE HAVE AN INACTIVE 5468 04:01:00,360 --> 04:01:04,640 ENZYME AS CONTROL. TO OVERCOME 5469 04:01:04,640 --> 04:01:07,680 ANY ARTIFACTS FROM THE NANOBODY 5470 04:01:07,680 --> 04:01:10,080 RECOGNITION ITSELF. SO PAUL 5471 04:01:10,080 --> 04:01:13,520 SHOWED THAT IF HE HAS THAT CK 2 5472 04:01:13,520 --> 04:01:15,480 ALPHA SOURCES CELLS AND PRESENCE 5473 04:01:15,480 --> 04:01:19,960 ACTIVE NANOBODY OGT WE CAN SEE 5474 04:01:19,960 --> 04:01:20,440 ELEVATED O-GlcNaCYLATION 5475 04:01:20,440 --> 04:01:22,720 AND THAT IS REDUCED AND REMOVED 5476 04:01:22,720 --> 04:01:24,880 OR DOESN'T OCCUR IN PRESENCE OF 5477 04:01:24,880 --> 04:01:27,640 THE INACTIVE NANOBODY OGA. IF WE 5478 04:01:27,640 --> 04:01:30,960 GLOBALLY ALTER THE 5479 04:01:30,960 --> 04:01:32,000 O-GlcNac PROTEOME WE DON'T 5480 04:01:32,000 --> 04:01:35,680 SEE AS SIGNIFICANT A INCREASE IN 5481 04:01:35,680 --> 04:01:36,480 O-GlcNaCYLATION OF THIS 5482 04:01:36,480 --> 04:01:38,960 TARGET PROTEIN COMPARED TO THE 5483 04:01:38,960 --> 04:01:40,480 NANOBE DI SYSTEM. BY CONTRAST 5484 04:01:40,480 --> 04:01:44,360 AND NOW IF WE COME IN AND LOAD A 5485 04:01:44,360 --> 04:01:47,960 LOT MORE PROTEINS WE CAN SEE 5486 04:01:47,960 --> 04:01:48,960 O-GlcNaCYLATION 5487 04:01:48,960 --> 04:01:51,120 ENDOGENOUSLY AS EXPECTED, 5488 04:01:51,120 --> 04:01:52,560 REMOVED BY GLOBAL 5489 04:01:52,560 --> 04:01:53,680 DEGLYCOSYLATION AND NOW MORE 5490 04:01:53,680 --> 04:01:56,440 TARGETED WAY WITH NANOBODY SPLIT 5491 04:01:56,440 --> 04:01:59,960 OGA SYSTEM THAT IS CONTROLLED 5492 04:01:59,960 --> 04:02:01,960 WITH THE CATALYTICALLY INACTIVE 5493 04:02:01,960 --> 04:02:03,600 VERSION. I DON'T HAVE TIME TO 5494 04:02:03,600 --> 04:02:07,360 SHOW ALL THE DATA OF PAUL'S 5495 04:02:07,360 --> 04:02:08,760 STUDY BUT HE SHOWED IN ADDITION 5496 04:02:08,760 --> 04:02:11,240 THE NANOBODY OGT SYSTEM IS 5497 04:02:11,240 --> 04:02:13,480 SELECTIVELY GLYCOSYLATING THE 5498 04:02:13,480 --> 04:02:15,600 SINGLE KNOWN GLYCOSIDE AND NOT 5499 04:02:15,600 --> 04:02:20,040 HITTING ANOTHER OFF TARGET 5500 04:02:20,040 --> 04:02:20,960 O-GglcNaCYLATION EVENT 5501 04:02:20,960 --> 04:02:23,080 IN THIS PARTICULAR PROTEIN, HE 5502 04:02:23,080 --> 04:02:25,360 CAN KNOCK IT OUT SELECTIVELY 5503 04:02:25,360 --> 04:02:27,160 WITH A SINGLE POINT MUTATION OR 5504 04:02:27,160 --> 04:02:29,440 A DOUBLE POINT MUTATION. AND 5505 04:02:29,440 --> 04:02:31,360 INTERESTINGLY IF WE TRY TO MAKE 5506 04:02:31,360 --> 04:02:34,800 A STABLE CYSTEINE GLYCOSYLATION 5507 04:02:34,800 --> 04:02:36,720 EVENT ENDOGENOUS OGA IS NOT 5508 04:02:36,720 --> 04:02:38,960 AFFECTING THAT. SO REALLY THIS 5509 04:02:38,960 --> 04:02:41,240 NANOBODY OGT OGA SYSTEM ALLOWS 5510 04:02:41,240 --> 04:02:44,600 US TO PURSUE THE NEXT QUESTION. 5511 04:02:44,600 --> 04:02:47,680 SO WITH THE NEXT QUESTION IS HOW 5512 04:02:47,680 --> 04:02:50,360 DOES THE O-GlcNac SHUN 5513 04:02:50,360 --> 04:02:53,480 EVENT ON CK 2 ALPHA ON THE 5514 04:02:53,480 --> 04:02:55,360 PROTEOME. WE DID 5515 04:02:55,360 --> 04:02:56,720 PHOSPHOPROTEOMICS. I WILL SHOW 5516 04:02:56,720 --> 04:02:59,720 YOU THE VOLCANO PLOT FOR 5517 04:02:59,720 --> 04:03:00,800 PHOSPHOSITE CHANGES IN THE 5518 04:03:00,800 --> 04:03:03,000 PRESENCE OF THE NANOBODY OGT 5519 04:03:03,000 --> 04:03:04,720 SYSTEM COMPARED TO THE 5520 04:03:04,720 --> 04:03:06,040 CATALYTICALLY INACTIVE FORM BUT 5521 04:03:06,040 --> 04:03:07,840 WE SEE BECAUSE WE HAVE MORE 5522 04:03:07,840 --> 04:03:10,520 GLYCOSYLATION WE ARE SEEING 5523 04:03:10,520 --> 04:03:13,560 ELEVATION IN SPECIFIC PHOSPHO 5524 04:03:13,560 --> 04:03:14,760 PHOSPHOPROTEINS WHEREAS WITH THE 5525 04:03:14,760 --> 04:03:17,680 NANOBODY OGA SYSTEM BECAUSE 5526 04:03:17,680 --> 04:03:21,800 O-GlcNac EVENTS ON CASEIN 5527 04:03:21,800 --> 04:03:23,560 KINASE 2 ALPHA WERE NOT THAT 5528 04:03:23,560 --> 04:03:25,240 HIGH NOW REDUCING IT DIDN'T LEAD 5529 04:03:25,240 --> 04:03:28,160 TO SUCH A LARGE PERTURBATION IN 5530 04:03:28,160 --> 04:03:33,360 THE PHOSPHOPROTEOME. THIS LED US 5531 04:03:33,360 --> 04:03:35,120 TO POSTULATE PHOSPHORYLATION ON 5532 04:03:35,120 --> 04:03:38,920 KNOWN CK 2 ALPHA SUBSTRATES HDAC 5533 04:03:38,920 --> 04:03:42,480 1 AND 2 THE 5534 04:03:42,480 --> 04:03:44,000 O-GglcNaCYLATION EVENT 5535 04:03:44,000 --> 04:03:46,560 IS AUGMENTING THE 5536 04:03:46,560 --> 04:03:49,520 PHOSPHOPROTEOMIC DATA AND 5537 04:03:49,520 --> 04:03:51,000 LEADING TO FOSTERED ACTIVITY. 5538 04:03:51,000 --> 04:03:53,160 THIS IS AN EXAMPLE WE ARE USING 5539 04:03:53,160 --> 04:03:54,360 THIS SYSTEM IN COLLABORATION 5540 04:03:54,360 --> 04:03:55,600 WITH OTHERS INCLUDING JOAN 5541 04:03:55,600 --> 04:03:58,840 WALKERS LAB A SPECIALIST IN 5542 04:03:58,840 --> 04:04:01,720 DROSOPHILA MODELS OF 5543 04:04:01,720 --> 04:04:02,600 NEURODEGENERATION AND 5544 04:04:02,600 --> 04:04:04,600 NEUROBIOLOGY, SPECIFICALLY 5545 04:04:04,600 --> 04:04:07,960 LOOKING AT HOW THE SYSTEM CAN 5546 04:04:07,960 --> 04:04:09,760 REGULATE SLEEP AND CIRCADIAN 5547 04:04:09,760 --> 04:04:14,360 BEHAVIOR. JIA NIU'S LAB BOSTON 5548 04:04:14,360 --> 04:04:15,520 COLLEGE, WE HAVE BEEN INTEGRATE 5549 04:04:15,520 --> 04:04:17,120 OUR TOO MANY SYSTEM WITH HIS TO 5550 04:04:17,120 --> 04:04:18,320 UNDERSTAND O-GlcNac IN THE 5551 04:04:18,320 --> 04:04:20,320 CONTEXT OF TRANSCRIPTIONAL 5552 04:04:20,320 --> 04:04:25,920 ACTIVATION AND EMILYN ALEJANDRO 5553 04:04:25,920 --> 04:04:28,000 WE MET THROUGH ONE OF THE NIH 5554 04:04:28,000 --> 04:04:30,360 MEETINGS SPECIFICALLY ON 5555 04:04:30,360 --> 04:04:31,320 O-GlcNac WHO HAS BEEN 5556 04:04:31,320 --> 04:04:34,440 PIONEERING USING THE TOOLS FOR 5557 04:04:34,440 --> 04:04:41,960 STUDYING DIABETES. AND THE 5558 04:04:41,960 --> 04:04:43,920 O-GlcNac IN O-GlcNac 5559 04:04:43,920 --> 04:04:45,960 TRANSFERASE IN BETA CELLS SO WE 5560 04:04:45,960 --> 04:04:48,360 USE THESE IN COLLABORATION OR 5561 04:04:48,360 --> 04:04:49,960 INDEPENDENT OF IT AND WE HAVE 5562 04:04:49,960 --> 04:04:52,360 PUT TOOLS IN AUTHORITY, IT'S THE 5563 04:04:52,360 --> 04:04:53,760 ONLY TOOLS WE HAVE ON ADD GENE 5564 04:04:53,760 --> 04:04:55,240 SO YOU CAN'T MISS THEM IF YOU 5565 04:04:55,240 --> 04:04:57,160 LOOK FOR THEM AND THESE TOOLS 5566 04:04:57,160 --> 04:04:59,440 HAVE BEEN DISTRIBUTED TO OF 5567 04:04:59,440 --> 04:05:03,160 WHICH OVER 50 SAMPLES HAVE BEEN 5568 04:05:03,160 --> 04:05:05,360 DISTRIBUTED. WITH THAT, I HOPE I 5569 04:05:05,360 --> 04:05:06,680 HAVE ABLE TO SHARE WITH YOU A 5570 04:05:06,680 --> 04:05:07,760 LITTLE BIT ABOUT WHAT WE HAVE 5571 04:05:07,760 --> 04:05:09,640 DONE OVER THE LAST FEW YEARS IN 5572 04:05:09,640 --> 04:05:12,040 THE -- AS PARTS OF THE COMMON 5573 04:05:12,040 --> 04:05:13,760 FUND, BOTH IN DEVELOPING THE 5574 04:05:13,760 --> 04:05:15,720 TOOLS AND NAVIGATING TO APPLY 5575 04:05:15,720 --> 04:05:16,880 THEM AND GOING TOWARDS THE 5576 04:05:16,880 --> 04:05:18,240 FUTURE WE ARE EXCITED TO BE ABLE 5577 04:05:18,240 --> 04:05:20,120 TO USE THESE AS WELL AS TO 5578 04:05:20,120 --> 04:05:20,840 CONTINUE TO DEVELOP NEW 5579 04:05:20,840 --> 04:05:24,960 VERSIONS. LAST BUT NOT LEAST I 5580 04:05:24,960 --> 04:05:25,960 HAVE TO THANK MY FANTASTIC LAB, 5581 04:05:25,960 --> 04:05:28,960 I HAVE HIGHLIGHTED MOST KEY 5582 04:05:28,960 --> 04:05:32,520 PLAYERS, AND THE PROJECTED AS I 5583 04:05:32,520 --> 04:05:35,360 HAVE GONE AND THANK YOU, VERY 5584 04:05:35,360 --> 04:05:35,680 MUCH. 5585 04:05:35,680 --> 04:05:44,200 [APPLAUSE] 5586 04:05:44,200 --> 04:05:46,400 >> OUR FINAL SPEAKER IN THE 5587 04:05:46,400 --> 04:05:47,480 O-GlcNac SESSION IS DR. 5588 04:05:47,480 --> 04:06:13,160 AI, UNIVERSITY OF VIRGINIA. 5589 04:06:13,160 --> 04:06:16,160 >> CAN YOU HEAR ME? OKAY. YES 5590 04:06:16,160 --> 04:06:18,880 IT'S WORKING. I JUST WANT TO 5591 04:06:18,880 --> 04:06:22,160 START MY PRESENTATION BY 5592 04:06:22,160 --> 04:06:26,960 THANKING PAM AND CAROL AND 5593 04:06:26,960 --> 04:06:29,720 CATHERINE FOR ORGANIZING THIS 5594 04:06:29,720 --> 04:06:32,920 WHOLE PROGRAM AND MY LAB 5595 04:06:32,920 --> 04:06:35,920 ACTUALLY WORKS WITH BIOSCIENCES 5596 04:06:35,920 --> 04:06:44,200 TO STUDY NUCLEOTIDE SUGARS. MY 5597 04:06:44,200 --> 04:06:46,880 RESEARCH STARTED WITH THIS 5598 04:06:46,880 --> 04:06:49,400 PARTICULAR IMPORTANT PROTEIN 5599 04:06:49,400 --> 04:06:51,360 GFP. THIS PROTEIN WAS DISCOVERED 5600 04:06:51,360 --> 04:06:55,040 IN LATE 1950s, MANY, MANY 5601 04:06:55,040 --> 04:06:58,680 YEARS AGO FROM JELLY FISH, FROM 5602 04:06:58,680 --> 04:07:01,480 THIS ORGANISM. AND JELLY FISH 5603 04:07:01,480 --> 04:07:05,000 HAS USED THIS PROTEIN FOR A 5604 04:07:05,000 --> 04:07:08,640 ENERGY TRANSFER SYSTEM FROM A 5605 04:07:08,640 --> 04:07:10,160 BIOLUMINESCENT PROTEIN AND THIS 5606 04:07:10,160 --> 04:07:12,360 PROTEIN HAS BEEN DEVELOPED AS A 5607 04:07:12,360 --> 04:07:15,760 MAJOR MARKER AND IN MY RESEARCH 5608 04:07:15,760 --> 04:07:19,960 WE USE AS A SCAFFOLD TO DEVELOP 5609 04:07:19,960 --> 04:07:21,040 BIORESEARCH. THE TOOLS IN 5610 04:07:21,040 --> 04:07:22,400 RESEARCH I WANT TO REMIND YOU 5611 04:07:22,400 --> 04:07:25,320 ABOUT CHEMISTRY, SO THIS 5612 04:07:25,320 --> 04:07:26,720 PARTICULAR PROTEIN IS INTERESTED 5613 04:07:26,720 --> 04:07:28,160 IN YOU PUT A GENE IN THIS 5614 04:07:28,160 --> 04:07:29,360 PROTEIN INTO LIVING CELLS, 5615 04:07:29,360 --> 04:07:34,520 LIVING ORGANISMS, SEVERAL 5616 04:07:34,520 --> 04:07:37,520 TYROSINE AND SEVERAL TYROSINE, 5617 04:07:37,520 --> 04:07:39,720 SORRY, SERINE TYROSINE IN THIS 5618 04:07:39,720 --> 04:07:43,120 PARTICULAR PROTEIN CAN UNDERGO 5619 04:07:43,120 --> 04:07:45,520 THIS SPONTANEOUS CHEMICAL 5620 04:07:45,520 --> 04:07:47,760 REACTION WHICH IS RESPONSIBLE 5621 04:07:47,760 --> 04:07:48,960 FOR THIS PARTICULAR PROTEIN, 5622 04:07:48,960 --> 04:07:52,200 LOOK AT STRUCTURE IN THE MIDDLE 5623 04:07:52,200 --> 04:07:56,760 OF THIS PROTEIN. THE IN MY 5624 04:07:56,760 --> 04:07:59,280 RESEARCH WE TRY TO USE PROTEIN 5625 04:07:59,280 --> 04:08:02,160 AS A MODULE, AS A SCAFFOLD TO 5626 04:08:02,160 --> 04:08:05,560 BRIDGE WITH SOME OTHER ELEMENT 5627 04:08:05,560 --> 04:08:08,920 WHICH CAN LEAD TO TARGET IN THIS 5628 04:08:08,920 --> 04:08:11,840 PARTICULAR PROJECT TO UDP SUGARS 5629 04:08:11,840 --> 04:08:14,760 SO WE HOPE THAT WE CAN BUILD A 5630 04:08:14,760 --> 04:08:17,760 HYBRID MOLECULE AND BIND WITH 5631 04:08:17,760 --> 04:08:20,280 THIS BINDING MODULE CAN MODULATE 5632 04:08:20,280 --> 04:08:21,320 FLUORESCENCE OF GREEN 5633 04:08:21,320 --> 04:08:25,160 FLUORESCENT PROTEIN. FOR THE 5634 04:08:25,160 --> 04:08:27,360 FIRST PART I WILL TALK ABOUT A 5635 04:08:27,360 --> 04:08:30,320 SPACE WHICH CAN SENSE UDP 5636 04:08:30,320 --> 04:08:32,480 GLCNAC. SO FOR THIS PARTICULAR 5637 04:08:32,480 --> 04:08:33,120 AUDIENCE I DON'T IMMEDIATE TO GO 5638 04:08:33,120 --> 04:08:35,920 THROUGH WHY THIS MOLECULE IS 5639 04:08:35,920 --> 04:08:36,960 IMPORTANT, BASICALLY IT IS A 5640 04:08:36,960 --> 04:08:40,160 MOLECULE WHICH BRIDGE METABOLITE 5641 04:08:40,160 --> 04:08:44,160 NUTRITION WITH SIGNAL DISEASE, 5642 04:08:44,160 --> 04:08:47,600 MY RESEARCH LOOK AT UDP GLCNAC 5643 04:08:47,600 --> 04:08:50,160 THIS PARTICULAR MOLECULE, WHICH 5644 04:08:50,160 --> 04:08:52,520 DISCUSSED IN THE PREVIOUS THREE 5645 04:08:52,520 --> 04:08:53,640 LECTURES, THE NEXT STEP TO 5646 04:08:53,640 --> 04:08:55,400 ENZYMES TO USE THIS MOLECULE AS 5647 04:08:55,400 --> 04:09:00,360 A SUBSTRATE TO MODIFY PROTEINS. 5648 04:09:00,360 --> 04:09:02,440 THEREFORE WE ARE STUDYING THIS 5649 04:09:02,440 --> 04:09:07,520 PARTICULAR SMALL MOLECULE, IN 5650 04:09:07,520 --> 04:09:10,000 LIVING CELLS AND INDICATOR WE 5651 04:09:10,000 --> 04:09:14,640 STUDY THIS BACTERIA UDP GLCNAC 5652 04:09:14,640 --> 04:09:16,200 TRANSFERASE AND THIS IS ENZYME 5653 04:09:16,200 --> 04:09:18,600 BASICALLY USED TO MODIFY LIPID 5654 04:09:18,600 --> 04:09:20,960 MOLECULE IN BACTERIA SO OUR HOPE 5655 04:09:20,960 --> 04:09:22,960 IS TO TRY TO GENERATE A HYBRID 5656 04:09:22,960 --> 04:09:25,560 MOLECULE WITH THIS GREEN 5657 04:09:25,560 --> 04:09:28,120 FLUORESCENT PROTEIN AND THIS UDP 5658 04:09:28,120 --> 04:09:30,200 GLCNAC BINDING PROTEIN AND WE 5659 04:09:30,200 --> 04:09:32,400 DID HOPE ACTUALLY TO GERMINATE 5660 04:09:32,400 --> 04:09:34,960 HYBRID MOLECULE TO CHANGE 5661 04:09:34,960 --> 04:09:37,360 FLUORESCENCE ON BINDING WITH 5662 04:09:37,360 --> 04:09:40,160 GLCNAC. I DON'T HAVE TIME TO 5663 04:09:40,160 --> 04:09:42,960 TELL DETAILS BUT BASICALLY 5664 04:09:42,960 --> 04:09:46,160 THROUGH COMPUTATIONAL ANALYSIS 5665 04:09:46,160 --> 04:09:47,920 AND EVOLUTION, WE EVENTUALLY 5666 04:09:47,920 --> 04:09:50,960 ENGINEER PROTEIN CALLED UGAG 5667 04:09:50,960 --> 04:09:53,600 WHICH CHANGE FLUORESCENCE BY 5668 04:09:53,600 --> 04:09:55,840 SEVEN FOLD IN RESPONSE TO UDP 5669 04:09:55,840 --> 04:10:00,800 GLCNAC. BASICALLY THE MIDDLE 5670 04:10:00,800 --> 04:10:02,360 SHOWS THE FLUORESCENCE 5671 04:10:02,360 --> 04:10:03,600 EXCITATION AND EMISSION OF THIS 5672 04:10:03,600 --> 04:10:07,360 PROTEIN AFTER BINDING WITH 5673 04:10:07,360 --> 04:10:09,320 GLCNAC, CHANGE CITATION WHICH 5674 04:10:09,320 --> 04:10:11,960 METRIC RESPONSE. THIS PROTEIN 5675 04:10:11,960 --> 04:10:14,200 SHOWED WHERE HIGH SPECIFICITY IN 5676 04:10:14,200 --> 04:10:16,400 RESPONSE TO UDP SUGARS FOR 5677 04:10:16,400 --> 04:10:22,360 EXAMPLE UDP GLCNAC HAS ONE 5678 04:10:22,360 --> 04:10:24,360 POSITION CHEMISTRY DIFFERENCE 5679 04:10:24,360 --> 04:10:28,200 BUT IT CAN DISTINGUISH GLCNAC 5680 04:10:28,200 --> 04:10:31,760 WITH GALNAC. YES DID NOR ASSAY 5681 04:10:31,760 --> 04:10:34,320 WE FIND -- MORE ASSAY WE FIND 5682 04:10:34,320 --> 04:10:39,520 THIS PROTEIN PART OF THE UDP. 5683 04:10:39,520 --> 04:10:42,520 THEN WE LOOK AT LITERATURE IN 5684 04:10:42,520 --> 04:10:46,040 ALL PAPERS FROM 2004 FROM 5685 04:10:46,040 --> 04:10:49,560 SUSAN'S WORK AND THIS PROTEIN IS 5686 04:10:49,560 --> 04:10:53,040 THE SAME AFFINITY. AND WE TRIED 5687 04:10:53,040 --> 04:10:54,920 -- IT WAS PROPOSED AS A 5688 04:10:54,920 --> 04:10:57,720 MECHANISM UDP AS A CANDIDATE 5689 04:10:57,720 --> 04:11:01,760 CELL REGULATOR OF THIS ENZYME 5690 04:11:01,760 --> 04:11:02,920 WHICH CAN (INAUDIBLE) ENZYME 5691 04:11:02,920 --> 04:11:05,160 ACTIVITY. WE TRIED HARD TO 5692 04:11:05,160 --> 04:11:08,480 ENGINEER ENZYME TO HAVE HIGH 5693 04:11:08,480 --> 04:11:13,840 SPECIFICITY DISTINGUISH UDP FROM 5694 04:11:13,840 --> 04:11:20,800 UDP GLCNAC. WE HAD SPS ALMOST NO 5695 04:11:20,800 --> 04:11:24,680 ACTIVITY WITH UDP GLCNAC BUT HAS 5696 04:11:24,680 --> 04:11:27,480 NEGATIVE CONTROL SENSOR. SO FOR 5697 04:11:27,480 --> 04:11:29,080 OUR SPECIFICITY, WE HAVE TO USE 5698 04:11:29,080 --> 04:11:32,720 THIS TWO SENSES TOGETHER. AND I 5699 04:11:32,720 --> 04:11:34,480 FOR THIS PARTICULAR EXPERIMENT 5700 04:11:34,480 --> 04:11:37,520 ON THIS SLIDE WE TRIED TO ADD 5701 04:11:37,520 --> 04:11:41,560 GLUCOSAMINE INTO THE CELL, SHOWS 5702 04:11:41,560 --> 04:11:44,640 THIS UDP GLCNAC SYSTEM PATHWAY 5703 04:11:44,640 --> 04:11:46,640 SO YOU ADD GLUCOSAMINE TO THE 5704 04:11:46,640 --> 04:11:49,600 CELL YOU CAN SEE IN THE UDP 5705 04:11:49,600 --> 04:11:51,880 GLCNAC SYNTHESIS AND AT THE SAME 5706 04:11:51,880 --> 04:11:54,680 TIME ADD UDP. GLUCOSAMINE IN THE 5707 04:11:54,680 --> 04:11:57,560 UPPER RIGHT CORNER, THE DATA IS 5708 04:11:57,560 --> 04:12:00,560 WORKING SO IT IS HARD TO CONTROL 5709 04:12:00,560 --> 04:12:03,520 THIS POINT. IN GLUCOSAMINE WE 5710 04:12:03,520 --> 04:12:07,120 SEE THIS INCREASE OF UDP GLCNAC 5711 04:12:07,120 --> 04:12:08,600 IN LIVING CELLS AND AT THE SAME 5712 04:12:08,600 --> 04:12:13,320 TIME WE SAW OBSERVE THE UXPS 5713 04:12:13,320 --> 04:12:15,600 DECREASE, BASICALLY THIS 5714 04:12:15,600 --> 04:12:17,280 INDICATES UTP CONCENTRATION 5715 04:12:17,280 --> 04:12:19,440 DECREASE IN LIVING CELLS BECAUSE 5716 04:12:19,440 --> 04:12:24,400 THIS CONSUMING PATHWAY. WE HAD 5717 04:12:24,400 --> 04:12:25,560 ANOTHER DATA MUTANT DATA 5718 04:12:25,560 --> 04:12:26,600 ESSENTIALLY WHICH SHOWS IN 5719 04:12:26,600 --> 04:12:32,640 RESPONSE TO UDP GLCNAC. THE 5720 04:12:32,640 --> 04:12:34,400 OTHER EXPERIMENT BY KNOCK DOWN 5721 04:12:34,400 --> 04:12:37,720 ENZYMES AND OVEREXPRESS ENZYMES 5722 04:12:37,720 --> 04:12:41,760 GFAP AND UAP AS EXPECTED THE 5723 04:12:41,760 --> 04:12:43,760 CONSENSUS OVEREXPRESS ENZYME AND 5724 04:12:43,760 --> 04:12:50,840 ALSO GENETIC KNOCK DOWN. MAYBE I 5725 04:12:50,840 --> 04:12:52,280 SHOULD HAVE USED THIS LASER 5726 04:12:52,280 --> 04:12:55,120 POINTER. USE THIS MOUSE I GUESS. 5727 04:12:55,120 --> 04:13:00,040 SO THE SAME THING OF THE WORK 5728 04:13:00,040 --> 04:13:03,920 ENZYME INHIBITOR LIKE 5729 04:13:03,920 --> 04:13:08,560 (INAUDIBLE) RESPONDED TO GFAP 5730 04:13:08,560 --> 04:13:11,840 INHIBITION. WE ONCE STUDY THIS 5731 04:13:11,840 --> 04:13:15,960 USE THIS TO STUDY UDP GLCNAC 5732 04:13:15,960 --> 04:13:19,960 CONCENTRATION IN MIN 6 CELLS. 5733 04:13:19,960 --> 04:13:22,360 BETA CELLS, CAN INHIBIT RESPONSE 5734 04:13:22,360 --> 04:13:24,760 TO GLUCOSE. SO WE TRY TO STUDY 5735 04:13:24,760 --> 04:13:27,800 WHETHER OR NOT UDP GLCNAC 5736 04:13:27,800 --> 04:13:28,920 CONCENTRATION IS SENSITIVE THE 5737 04:13:28,920 --> 04:13:32,560 GLUCOSE IN BETA CELLS, BASICALLY 5738 04:13:32,560 --> 04:13:36,320 SHOWS WE FIRST VALIDATED WASN'T 5739 04:13:36,320 --> 04:13:40,240 WORKING BETA CELLS BY LOOKING AT 5740 04:13:40,240 --> 04:13:42,240 ADDITIONAL GLUCOSAMINE. SO THEN 5741 04:13:42,240 --> 04:13:44,960 WE STARTED WITH GLUCOSE TO OUR 5742 04:13:44,960 --> 04:13:48,120 SURPRISE THE GLUCOSE 5743 04:13:48,120 --> 04:13:50,160 CONCENTRATION BY 2 MILLIMOLAR TO 5744 04:13:50,160 --> 04:13:52,600 25 MILLIMOLAR CONCENTRATION, 5745 04:13:52,600 --> 04:13:53,800 DIDN'T CHANGE GLCNAC 5746 04:13:53,800 --> 04:13:54,800 CONCENTRATION TOO MUCH. AND IN 5747 04:13:54,800 --> 04:13:57,560 ORDER THE STAY LIKE IN THE 5748 04:13:57,560 --> 04:14:01,040 GLUCOSE CHAIN MODULATION UDP 5749 04:14:01,040 --> 04:14:05,360 GLCNAC WE HAD TO USE ZERO 5750 04:14:05,360 --> 04:14:06,280 MILIMISSOURI LAR AND OBSERVE -- 5751 04:14:06,280 --> 04:14:07,760 MILLIMOLAR AND OBSERVE THE 5752 04:14:07,760 --> 04:14:09,440 CHANGE. THIS WAS KIND OF A QUITE 5753 04:14:09,440 --> 04:14:12,960 A SURPRISE BUT BASICALLY OUR 5754 04:14:12,960 --> 04:14:17,880 STUDY SHOWS BEAUTIFUL GLCNAC 5755 04:14:17,880 --> 04:14:19,840 LEVEL CELLS IS -- MIN 6 CELLS IS 5756 04:14:19,840 --> 04:14:23,240 NOT SENSITIVE TO GLUCOSE 5757 04:14:23,240 --> 04:14:25,840 CONCENTRATION. IT IS QUITE 5758 04:14:25,840 --> 04:14:28,160 REGULATED. MEANWHILE WE ALSO 5759 04:14:28,160 --> 04:14:31,680 STUDY ENGINEERED FLUORESCENT 5760 04:14:31,680 --> 04:14:36,280 ORIGINATING GLCNAC AND THIS WILL 5761 04:14:36,280 --> 04:14:42,600 IS ALPHA RESPONSE. UDP GLCNAC 5762 04:14:42,600 --> 04:14:44,760 INCREASE, TWO DECREASE, 2 5763 04:14:44,760 --> 04:14:46,520 MILLIMOLAR BUT A DIFFERENT 5764 04:14:46,520 --> 04:14:48,680 SPECIFICITY, IT SEEMS AT LEAST 5765 04:14:48,680 --> 04:14:55,560 FOR THE MOLECULE ONLY UDP GLCNAC 5766 04:14:55,560 --> 04:14:58,400 AND GALNAC INDUCE A RESPONSE 5767 04:14:58,400 --> 04:15:01,560 HERE. WE WERE ABLE TO LOCALIZE 5768 04:15:01,560 --> 04:15:06,360 TO HERE COMPATIBLE WITH MORE UDP 5769 04:15:06,360 --> 04:15:08,480 GLCNAC AND OBSERVE 2 DG AND 5770 04:15:08,480 --> 04:15:11,920 GLUCOSAMINE INDUCED UDP GLCNAC 5771 04:15:11,920 --> 04:15:13,080 CONCENTRATION CHANGE INDUCE 5772 04:15:13,080 --> 04:15:13,960 COMPARTMENT WITH HIGH 5773 04:15:13,960 --> 04:15:17,560 CONCENTRATION OF UDP GLCNAC. IN 5774 04:15:17,560 --> 04:15:21,520 ADDITION TO UDP GLCNAC WE 5775 04:15:21,520 --> 04:15:23,440 ENGINEERED A SENSE OF GLUCOSE. 5776 04:15:23,440 --> 04:15:26,400 UDP GLUCOSE IS MOLECULE, 5777 04:15:26,400 --> 04:15:28,320 IMPORTANT FOR GLUCOSE 5778 04:15:28,320 --> 04:15:30,120 CONCENTRATION REGULATION IN OUR 5779 04:15:30,120 --> 04:15:33,520 BODY. ENERGY STORAGE. SO THAT IS 5780 04:15:33,520 --> 04:15:37,040 UDP GLUCOSE IS MOLECULE 5781 04:15:37,040 --> 04:15:41,720 INTERMEDIATE TO STORE GROUP TO 5782 04:15:41,720 --> 04:15:44,880 UNIFORM GLYCOGEN. THIS IS VERY 5783 04:15:44,880 --> 04:15:47,760 IMPORTANT FOR GLUCOSE 5784 04:15:47,760 --> 04:15:52,240 CONCENTRATION REGULATION IN 5785 04:15:52,240 --> 04:15:55,840 BLOOD. WE INHIBITOR OF GFP TO 5786 04:15:55,840 --> 04:16:02,280 PROTEIN WE CALL UGP, IT IS UDP 5787 04:16:02,280 --> 04:16:04,440 GLUCOSE PIE ROW PHOSPHORYLATION. 5788 04:16:04,440 --> 04:16:07,880 SO ENZYMES HAVE O NO CATALYTIC 5789 04:16:07,880 --> 04:16:10,920 ACTIVITY AND WE MADE THIS IN THE 5790 04:16:10,920 --> 04:16:12,560 MOLECULE RESPONDING TO UDP 5791 04:16:12,560 --> 04:16:14,320 GLUCOSE. WE STARTED WITH 5792 04:16:14,320 --> 04:16:17,320 MODULATION WITH K ENZYMES 5793 04:16:17,320 --> 04:16:19,560 BASICALLY KNOCK DOWN PGM AND 5794 04:16:19,560 --> 04:16:24,400 KNOCK DOWN UGP AND WE CAN 5795 04:16:24,400 --> 04:16:26,240 OBSERVE RESPONSE AND ALSO 5796 04:16:26,240 --> 04:16:31,800 OVEREXPRESS CO-ENZYME, OBSERVE 5797 04:16:31,800 --> 04:16:33,200 ALSO RESPONSE. WE UNTIL RECENTLY 5798 04:16:33,200 --> 04:16:40,160 TRIED TO EXPRESS THE SAME IN A 5799 04:16:40,160 --> 04:16:43,120 ZEBRAFISH ORGANISM. WHICH CAN 5800 04:16:43,120 --> 04:16:45,280 CONFER EXPLORATION OF PROTEIN IN 5801 04:16:45,280 --> 04:16:48,320 MUSCLE CELLS AND WE CAN OBSERVE 5802 04:16:48,320 --> 04:16:52,360 SINGLE CELL RESOLUTION WE ADD 5803 04:16:52,360 --> 04:16:56,760 MOLECULE FOR EPINEPHRIN MOLECULE 5804 04:16:56,760 --> 04:16:58,520 ON THE RIGHT SIDE, THIS CAN 5805 04:16:58,520 --> 04:17:01,000 INCREASE CMP CONCENTRATION IN 5806 04:17:01,000 --> 04:17:02,880 CELLS AND THIS ACTIVITY -- 5807 04:17:02,880 --> 04:17:05,520 PROTEIN KINASE A WHICH DO TWO 5808 04:17:05,520 --> 04:17:08,560 THINGS WITH THIS UDP GLUCOSE, 5809 04:17:08,560 --> 04:17:14,040 ONE IS INCREASE GLYCOGEN 5810 04:17:14,040 --> 04:17:15,680 CONVERSION TO GLUCOSE 1 5811 04:17:15,680 --> 04:17:18,920 PHOSPHATE. AND THE OTHER ONE IS 5812 04:17:18,920 --> 04:17:22,560 INHIBITED CENSUS, THIS MOLECULE 5813 04:17:22,560 --> 04:17:25,160 WE ARE INCREASE UDP GLUCOSE 5814 04:17:25,160 --> 04:17:26,160 CONCENTRATION, I FORGOT TO 5815 04:17:26,160 --> 04:17:29,560 MENTION THIS MOLECULE IS THIS 5816 04:17:29,560 --> 04:17:31,840 SENSOR OUR SENSOR IS HAVE 5817 04:17:31,840 --> 04:17:33,960 REVERSE RESPONSE SO WHEN GLUCOSE 5818 04:17:33,960 --> 04:17:35,680 INCREASE WE OBSERVE THE DECREASE 5819 04:17:35,680 --> 04:17:39,160 IN FLUORESCENCE. SO WE OBSERVE 5820 04:17:39,160 --> 04:17:42,120 THE MOLECULE IN ZEBRAFISH WE 5821 04:17:42,120 --> 04:17:45,640 OBSERVE INCREASE OF UDP GLUCOSE. 5822 04:17:45,640 --> 04:17:47,160 ABOUT 30 MINUTES THIS RESPONSE 5823 04:17:47,160 --> 04:17:50,360 IS REVERSED SO BASICALLY WE 5824 04:17:50,360 --> 04:17:54,960 COULD OBSERVE OF THE 5825 04:17:54,960 --> 04:17:56,880 STIMULATION, -- EPINEPHRINE 5826 04:17:56,880 --> 04:17:58,680 STIMULATION. WE OBSERVE SIGNAL 5827 04:17:58,680 --> 04:18:02,280 DYNAMICS TO INCREASE UDP 5828 04:18:02,280 --> 04:18:05,560 GLYCOGEN SYNTHESIS AND INCREASE 5829 04:18:05,560 --> 04:18:06,800 WITH RUBBING GLYCOGEN LYSIS AND 5830 04:18:06,800 --> 04:18:10,360 IN 30 MINUTES WITH THIS COULD BE 5831 04:18:10,360 --> 04:18:11,920 GLUCOSE CITRATEED FROM THE 5832 04:18:11,920 --> 04:18:13,840 MUSCLE CELL OUTSIDE OF THE CELL 5833 04:18:13,840 --> 04:18:16,680 OR MECHANISM WE DON'T QUITE 5834 04:18:16,680 --> 04:18:18,520 UNDERSTAND BUT TERMINATE THIS 5835 04:18:18,520 --> 04:18:19,480 SIGNALING EVENT AFTER 30 5836 04:18:19,480 --> 04:18:26,360 MINUTES. TO SUMMARIZE WE 5837 04:18:26,360 --> 04:18:31,960 ENGINEER GENETIC ENCODED SENSE 5838 04:18:31,960 --> 04:18:35,280 IN POLYGLYCOSIDE AND ENGINEERED 5839 04:18:35,280 --> 04:18:39,560 GENETICALLY ENCODED UDP GLCNAC, 5840 04:18:39,560 --> 04:18:41,200 AND FOLLOWED WITH CELLS 5841 04:18:41,200 --> 04:18:42,160 INHIBITORS NOT BOUND TO 5842 04:18:42,160 --> 04:18:45,520 OVEREXPRESSION OF ENZYMES AND WE 5843 04:18:45,520 --> 04:18:48,920 MONITOR UDP GLCNAC LEVELS IN 5844 04:18:48,920 --> 04:18:50,040 PANCREATIC BETA CELLS AND 5845 04:18:50,040 --> 04:18:55,040 SUGGEST GLUCOSE METABOLISM HPV 5846 04:18:55,040 --> 04:18:56,840 IS TIGHTLY REGULATED IN BETA 5847 04:18:56,840 --> 04:19:01,480 CELLS AND EXPRESS UDP GLUCOSE IN 5848 04:19:01,480 --> 04:19:03,160 ZEBRAFISH AND OBSERVE UDP 5849 04:19:03,160 --> 04:19:05,160 GLUCOSE CONCENTRATION CHANGE IN 5850 04:19:05,160 --> 04:19:07,520 RESPONSE TO EPINEPHRINE SO WE 5851 04:19:07,520 --> 04:19:09,960 THINK THIS EVENTUALLY HELP US 5852 04:19:09,960 --> 04:19:11,080 UNDERSTAND DISEASE SUCH AS 5853 04:19:11,080 --> 04:19:15,160 DIABETES FOR EXAMPLE. AND THEN 5854 04:19:15,160 --> 04:19:21,040 WE -- OUR RESEARCH IS SUPPORTED 5855 04:19:21,040 --> 04:19:26,760 BY GRANTS. THE WORK WE PRESENTED 5856 04:19:26,760 --> 04:19:29,640 TODAY WERE DONE MOSTLY BY THESE 5857 04:19:29,640 --> 04:19:32,560 POST DOCS ZEFAL LI, WHO IS HERE 5858 04:19:32,560 --> 04:19:34,360 AND HAS A POSTER AND YOU CAN ASK 5859 04:19:34,360 --> 04:19:36,920 HIM MORE DETAILS, SOMETIMES MORE 5860 04:19:36,920 --> 04:19:38,760 DETAILS I DON'T KNOW. AND 5861 04:19:38,760 --> 04:19:44,600 ANOTHER POST-DOC JING ZHANG 5862 04:19:44,600 --> 04:19:47,320 COMPLETED THIS WORK. IF YOU 5863 04:19:47,320 --> 04:19:48,360 DON'T FIND ENERGY SEND ME AN 5864 04:19:48,360 --> 04:19:51,960 EMAIL. I'M HAPPY TO SHARE MY 5865 04:19:51,960 --> 04:19:54,800 MATERIAL. ADD GENE ABOUT 100,000 5866 04:19:54,800 --> 04:20:00,480 SO YEAH. THANK YOU. 5867 04:20:00,480 --> 04:20:03,800 [APPLAUSE] 5868 04:20:03,800 --> 04:20:05,840 >> THAT CONCLUDES OUR 5869 04:20:05,840 --> 04:20:07,840 O-GlcNac SESSION NOW I 5870 04:20:07,840 --> 04:20:10,280 HAND IT TO NATASHA FOR 5871 04:20:10,280 --> 04:20:12,400 GLYCOPROTEOMICS. THE FIRST 5872 04:20:12,400 --> 04:20:14,360 SPEAKER IN THE GLYCOPROTEOMICS 5873 04:20:14,360 --> 04:20:16,600 SESSION IS DR. ROBERT WOODS CCRC 5874 04:20:16,600 --> 04:20:19,560 UNIVERSITY OF GEORGIA TALKING 5875 04:20:19,560 --> 04:20:22,160 ABOUT GLYCAN STRUCTURE PRE-AND 5876 04:20:22,160 --> 04:20:30,000 POST DEPOSITION IN THE PDP. 5877 04:20:30,000 --> 04:20:32,440 >> LIKE ALL SPEAKERS I'M 5878 04:20:32,440 --> 04:20:34,920 GENUINELY WANTING TO THANK THE 5879 04:20:34,920 --> 04:20:37,480 NIH PAM, CARL, DOUG, FOR THE 5880 04:20:37,480 --> 04:20:39,520 UNFAILING SUPPORT OF THE COMMON 5881 04:20:39,520 --> 04:20:40,560 FUND INITIATIVE AND EVEN BEFORE 5882 04:20:40,560 --> 04:20:42,960 THAT. AS A TOOL DEVELOPER I CAST 5883 04:20:42,960 --> 04:20:47,080 MY MIND BACK TO THE YEARS BEFORE 5884 04:20:47,080 --> 04:20:50,880 WE HAD THE TOOL INITIATIVE AND 5885 04:20:50,880 --> 04:20:51,600 BOY IT WAS BLEAK AND DIFFICULT 5886 04:20:51,600 --> 04:20:52,720 TO GET FUNDED SO THIS WAS 5887 04:20:52,720 --> 04:20:56,800 TREMENDOUS FOR US. I WILL TALK 5888 04:20:56,800 --> 04:20:58,560 TODAY ABOUT AN INITIATIVE WE HAD 5889 04:20:58,560 --> 04:21:08,280 THAT WAS TOGETHER WITH JASMINE 5890 04:21:08,280 --> 04:21:09,520 YOUNG, THIS WAS ONE OF THE BIG 5891 04:21:09,520 --> 04:21:11,760 PROBLEMS WE WERE AWARE OF IN THE 5892 04:21:11,760 --> 04:21:14,160 PDP. WHEN I SAY WE ALL, I MEAN 5893 04:21:14,160 --> 04:21:16,600 ALL THE OF US OBSESSED WITH 3-D 5894 04:21:16,600 --> 04:21:19,200 STRUCTURE AS CATH RIB SAID. -- 5895 04:21:19,200 --> 04:21:21,280 CATHERINE SAID. SO TURNS OUT AS 5896 04:21:21,280 --> 04:21:23,760 OF MAY -- SO THIS MONTH THERE 5897 04:21:23,760 --> 04:21:26,440 ARE ALMOST 190,000 MACRO 5898 04:21:26,440 --> 04:21:30,480 MOLECULAR STRUCTURES IN THE PDB. 5899 04:21:30,480 --> 04:21:33,560 USING THIS PROGRAM WE KNOW 5900 04:21:33,560 --> 04:21:35,680 CARBOHYDRATES ARE AVAILABLE IN 5901 04:21:35,680 --> 04:21:38,160 19,000 PDP OR 10% OF THE PDB. 5902 04:21:38,160 --> 04:21:41,360 THIS WASN'T KNOWABLE UNTIL WE 5903 04:21:41,360 --> 04:21:42,880 DID THIS. IT WAS ESTIMATED 5904 04:21:42,880 --> 04:21:45,200 VARIOUS WAYS BUT THE PROBLEM WAS 5905 04:21:45,200 --> 04:21:49,520 THAT THE PDB HAD NO STANDARD 5906 04:21:49,520 --> 04:21:50,760 NOMENCLATURE AND MANY ERRORS 5907 04:21:50,760 --> 04:21:52,680 MANY THE THE NOMENCLATURE SO YOU 5908 04:21:52,680 --> 04:21:54,440 COULDN'T SEARCH AND FIND SUGARS. 5909 04:21:54,440 --> 04:22:00,840 AND EVEN TODAY, THERE'S SEARCH 5910 04:22:00,840 --> 04:22:01,760 INTERFACES IS EXTRAORDINARILY 5911 04:22:01,760 --> 04:22:03,640 LIMITED SO WE CREATED OUR OWN 5912 04:22:03,640 --> 04:22:06,560 INTERFACE FOR RESEARCH IN PDB. 5913 04:22:06,560 --> 04:22:08,960 BECAUSE OF LACK OF ATTENTION TO 5914 04:22:08,960 --> 04:22:11,200 CARBOHYDRATES EARLY ON IN THE 5915 04:22:11,200 --> 04:22:13,160 PDB DEPOSITION THERE ARE LOTS OF 5916 04:22:13,160 --> 04:22:14,800 INCONSISTENCIES THAT PUSH THE 5917 04:22:14,800 --> 04:22:16,600 PDB TO WORK TOGETHER WITH US TO 5918 04:22:16,600 --> 04:22:18,600 DO WHAT THEY CALL REMEDIATION. 5919 04:22:18,600 --> 04:22:22,400 WHICH IS GOING BACK THROUGH THE 5920 04:22:22,400 --> 04:22:24,080 DATA, FINDING ERRORS IN 5921 04:22:24,080 --> 04:22:25,760 STRUCTURES WHICH THEY DON'T 5922 04:22:25,760 --> 04:22:28,360 CORRECT BUT FLAG AND ERROR MS. 5923 04:22:28,360 --> 04:22:31,000 NOMENCLATURE AND SO FORTH. THIS 5924 04:22:31,000 --> 04:22:32,080 -- UNTIL THEY DID THIS THIS 5925 04:22:32,080 --> 04:22:33,760 CAUSED A LOT OF CONFUSION. 5926 04:22:33,760 --> 04:22:35,760 BECAUSE WHEN STRUCTURE IS 5927 04:22:35,760 --> 04:22:37,000 DEPOSITED PEOPLE LIKE ME WHO DO 5928 04:22:37,000 --> 04:22:38,360 MODELING AND RETRIEVE THE 5929 04:22:38,360 --> 04:22:39,120 STRUCTURE THINK IT IS RIGHT 5930 04:22:39,120 --> 04:22:40,560 BECAUSE IT WAS PUT THERE. AND I 5931 04:22:40,560 --> 04:22:42,440 TELL YOU WHAT, IF WE ARE 5932 04:22:42,440 --> 04:22:43,520 MODELING PEOPLE TRYING TO 5933 04:22:43,520 --> 04:22:46,400 CONVINCE PEOPLE WE ARE WRITING 5934 04:22:46,400 --> 04:22:47,440 CRYSTAL STRUCTURE DRAWING THAT 5935 04:22:47,440 --> 04:22:48,760 IS HARD BATTLE SO THIS IS A WAY 5936 04:22:48,760 --> 04:22:52,280 TO OBJECTIVELY LOOK AT THAT AND 5937 04:22:52,280 --> 04:22:52,880 FIGURE WHAT IS RIGHT AND WRONG 5938 04:22:52,880 --> 04:22:54,560 AND HELPS WHEN YOU ARE TRYING TO 5939 04:22:54,560 --> 04:22:56,160 DO THINGS LIKE MOLECULAR 5940 04:22:56,160 --> 04:22:57,400 MODELING OR PREDICTION OF 5941 04:22:57,400 --> 04:23:03,000 STRUCTURE OR THAT SORT OF THING. 5942 04:23:03,000 --> 04:23:08,720 THERE'S TENS OF THOUSANDS OF 5943 04:23:08,720 --> 04:23:12,160 COORDINATE SETS OF SUGARS IN THE 5944 04:23:12,160 --> 04:23:12,960 PDB THAT'S HUNDREDS OF THOUSANDS 5945 04:23:12,960 --> 04:23:15,520 OF HOURS OF CRYSTALLOGRAPHERS 5946 04:23:15,520 --> 04:23:17,440 TIME WHEN IT WASN'T AUTOMATED 5947 04:23:17,440 --> 04:23:18,680 SOLUTION OF CRYSTAL STRUCTURE, 5948 04:23:18,680 --> 04:23:20,360 THIS IS MANUAL WORK TO GET THESE 5949 04:23:20,360 --> 04:23:22,560 IN THERE THERE. AND U YOU COULDN'T 5950 04:23:22,560 --> 04:23:23,360 FIND THEM, IT WAS REALLY A 5951 04:23:23,360 --> 04:23:29,760 PROBLEM. SO PDB WENT FORWARD 5952 04:23:29,760 --> 04:23:30,720 WITH REMEDIATION, TOOK WHAT WE 5953 04:23:30,720 --> 04:23:32,360 DEVELOPED AND USED IT TO FIND 5954 04:23:32,360 --> 04:23:35,080 MISTAKES AND WENT THROUGH AND 5955 04:23:35,080 --> 04:23:38,880 14,000 FILES WERE REMEDIATED. 5956 04:23:38,880 --> 04:23:42,600 IF I CAN GO BACK. 18,000 EXIST. 5957 04:23:42,600 --> 04:23:47,360 SO 14 OUT OF 18 HAD PROBLEMS. 5958 04:23:47,360 --> 04:23:53,160 THAT'S A THING TO THINK ABOUT. 5959 04:23:53,160 --> 04:23:56,120 COUPLE OF THINGS I HEME HAPPY 5960 04:23:56,120 --> 04:23:57,160 WITH THEY SCRIBED 5961 04:23:57,160 --> 04:23:59,160 OLIGOSACCHARIDES WITH POLYMERS 5962 04:23:59,160 --> 04:24:00,920 OF MONOSACCHARIDES. IT WASN'T 5963 04:24:00,920 --> 04:24:02,480 THAT WAY INITIALLY, THEY WERE 5964 04:24:02,480 --> 04:24:06,720 JUST CRYSTALLOGRAPHER WE CALLED 5965 04:24:06,720 --> 04:24:08,560 THEM MAN 3. IT WAS MAN ALPHA 1, 5966 04:24:08,560 --> 04:24:10,160 2, BLAH BLAH, JUST CALL IT, IT 5967 04:24:10,160 --> 04:24:15,280 IS A LIGAND. THEY INTRODUCED 5968 04:24:15,280 --> 04:24:17,320 3-DS AND FGG. THIS CAME OUT OF 5969 04:24:17,320 --> 04:24:20,960 THIS WORK. YOU CAN SEE HERE THE 5970 04:24:20,960 --> 04:24:22,560 STANDARD 2 DS FG RECOMMENDATION 5971 04:24:22,560 --> 04:24:24,480 AND THEN TO THE RIGHT OF IT THE 5972 04:24:24,480 --> 04:24:26,200 UPPER RIGHT CORNER, THE 5973 04:24:26,200 --> 04:24:28,200 TRADITIONAL 3-D, PROFOUND 5974 04:24:28,200 --> 04:24:30,880 DIFFERENCES. UNLESS YOU ARE 5975 04:24:30,880 --> 04:24:32,960 OBSESSED WITH 3-D STRUCTURE 5976 04:24:32,960 --> 04:24:34,120 CARBOHYDRATES YOU CANNOT LOOK AT 5977 04:24:34,120 --> 04:24:35,920 THE 3-D STRUCTURE UPPER RIGHT 5978 04:24:35,920 --> 04:24:39,360 AND SEE WHERE FUCOSE IS. WE 5979 04:24:39,360 --> 04:24:40,520 THOUGHT THERE MUST BE A BETTER 5980 04:24:40,520 --> 04:24:43,520 WAY SO WE TOOK 2 DS AND FG AND 5981 04:24:43,520 --> 04:24:46,720 PUT IN AS 3-D ICONS OR 3-D 5982 04:24:46,720 --> 04:24:49,360 SYMBOLS. AND I THINK THIS IS AS 5983 04:24:49,360 --> 04:24:50,800 REVOLUTIONARY AS USING RIBBON 5984 04:24:50,800 --> 04:24:53,360 STRUCTURE FOR PROTEINS. IT HELPS 5985 04:24:53,360 --> 04:24:54,560 YOU IMMEDIATELY SEE THE KEY 5986 04:24:54,560 --> 04:24:57,160 THINGS GOING ON AND ONE OF THE 5987 04:24:57,160 --> 04:24:58,240 THANKS POINTS OUT IS THIS 5988 04:24:58,240 --> 04:25:00,280 WONDERFUL LITTLE LESS THAN FG 5989 04:25:00,280 --> 04:25:01,960 NOTATION THAT IS SYMMETRIC IS 5990 04:25:01,960 --> 04:25:03,760 NOT REPRESENTATIVE OF MOLECULAR 5991 04:25:03,760 --> 04:25:04,920 STRUCTURE. THESE ARE NOT 5992 04:25:04,920 --> 04:25:08,960 SYMMETRIC MOLECULES. SO ONE OF 5993 04:25:08,960 --> 04:25:10,360 THE TOOLS WE DEVELOPED IS THIS 5994 04:25:10,360 --> 04:25:12,360 SEARCHING FOR GLYCANS IN THE PDB 5995 04:25:12,360 --> 04:25:15,160 SO GO O THIS WEB SITES GLIA 5996 04:25:15,160 --> 04:25:17,960 CONDITION.ORG YOU CAN DO THIS. 5997 04:25:17,960 --> 04:25:20,920 AS LARRY TABAK SAID ON THE FIRST 5998 04:25:20,920 --> 04:25:23,000 DAY, WE ARE SUPPOSED TO BE 5999 04:25:23,000 --> 04:25:24,160 DEVELOPING TOOLS FOR 6000 04:25:24,160 --> 04:25:26,200 NON-SPECIALISTS. AND THAT IS 6001 04:25:26,200 --> 04:25:27,680 EXACTLY THE WAY WE DID THIS. WE 6002 04:25:27,680 --> 04:25:29,560 TOOK OUR OLD WEBSITE AND SAID 6003 04:25:29,560 --> 04:25:32,480 NO, NO, THAT IS FOR SPECIALISTS 6004 04:25:32,480 --> 04:25:36,200 WHAT COULD GYCO DO? NOBODY LIKES 6005 04:25:36,200 --> 04:25:37,480 GETTING INSURANCE, SO YOU GO TO 6006 04:25:37,480 --> 04:25:39,760 WEBSITE SEEMS EASY, WE THOUGHT 6007 04:25:39,760 --> 04:25:42,160 MUST BE EQUIVALENT THING FOR US 6008 04:25:42,160 --> 04:25:44,160 SO WE TRIED TO KEEP THAT MIND 6009 04:25:44,160 --> 04:25:45,120 SET OF HOW TO DO. 6010 04:25:45,120 --> 04:25:48,160 THIS AND WE DECIDED BY 6011 04:25:48,160 --> 04:25:49,560 INTERROGATION BASED BUTTONS SO 6012 04:25:49,560 --> 04:25:50,760 WHEN A NON-SPECIALIST LANDS 6013 04:25:50,760 --> 04:25:52,320 THERE AND SAYS WHAT ARE THE 6014 04:25:52,320 --> 04:25:53,880 SHAPE OF MY GLYCANS? THAT IS 6015 04:25:53,880 --> 04:25:56,640 WHAT I WANT TO KNOW. SO IT IS 6016 04:25:56,640 --> 04:25:57,480 MORE OBVIOUS THAN HOW IT WAS 6017 04:25:57,480 --> 04:26:01,760 BEFORE. WHEN YOU DO THIS, THERE 6018 04:26:01,760 --> 04:26:03,960 IS MULTIPLE WAYS TO FIND GLYCANS 6019 04:26:03,960 --> 04:26:06,960 IN THE PDB NOW. WE ARE GOING TO 6020 04:26:06,960 --> 04:26:08,960 -- I WILL SHOW YOU A POINT AND 6021 04:26:08,960 --> 04:26:10,200 CLICK I WISH I COULD DO IT LIVE 6022 04:26:10,200 --> 04:26:12,120 BUT WE WILL DO IT THIS WAY. BUT 6023 04:26:12,120 --> 04:26:17,120 GO TO GLYCAN.ORG DOES HE 6024 04:26:17,120 --> 04:26:19,560 UNDERSTAND THIS FOR YOURSELF. IT 6025 04:26:19,560 --> 04:26:21,040 WILL POP UP THIS POINT AND CLICK 6026 04:26:21,040 --> 04:26:22,440 SCREEN. NOTICE A COUPLE OF 6027 04:26:22,440 --> 04:26:24,160 THINGS, ONE YOU SEE THE LITTLE 6028 04:26:24,160 --> 04:26:27,360 STAR, WILD CARD, THAT IS TO HELP 6029 04:26:27,360 --> 04:26:28,600 YOU SEARCH, ANYTHING IN FRONT 6030 04:26:28,600 --> 04:26:29,720 CAN BE WHATEVER YOU WANT JUST 6031 04:26:29,720 --> 04:26:32,920 LIKE YOU THINK A WILD CARD DOES. 6032 04:26:32,920 --> 04:26:35,760 SO THIS SEARCH FINDS ANYTHING 6033 04:26:35,760 --> 04:26:39,160 WITH -- NS TO GLUCOSAMINE IN 6034 04:26:39,160 --> 04:26:40,520 PYRANOSE FORM. IT IS FAST AND 6035 04:26:40,520 --> 04:26:43,160 THE REASON IT IS FAST IS WE 6036 04:26:43,160 --> 04:26:46,960 WROTE A NEW PDB. WE CREATED A 6037 04:26:46,960 --> 04:26:49,040 DATABASE THAT IS CUSTOMIZED FOR 6038 04:26:49,040 --> 04:26:51,120 STORING DATA FOR SUGARS THAT WE 6039 04:26:51,120 --> 04:26:52,640 PULL OUT OF THE PDB. WE STORE 6040 04:26:52,640 --> 04:26:55,360 ALL THE DATA IN ANY ENTRY THAT 6041 04:26:55,360 --> 04:27:00,440 HAS A SUGAR. THAT MEANS WE CAN 6042 04:27:00,440 --> 04:27:01,960 CREATE OUR OWN WAY TO STORE THE 6043 04:27:01,960 --> 04:27:03,280 DATA IN OUR OWN WAY TO SEARCH 6044 04:27:03,280 --> 04:27:05,440 THE DATA. WE UPDATE THIS 6045 04:27:05,440 --> 04:27:06,400 PERIODICALLY SO IT IS ALWAYS UP 6046 04:27:06,400 --> 04:27:10,440 TO DATE. WHEN YOU CLICK ON IT 6047 04:27:10,440 --> 04:27:12,640 RIGHT AWAY, IT SEARCHES 95,000 6048 04:27:12,640 --> 04:27:15,760 ENTRIES WITH SUGARS. IT 6049 04:27:15,760 --> 04:27:17,400 PROVIDES BACK UNIFORM 6050 04:27:17,400 --> 04:27:18,840 NOMENCLATURE WHAT THE SUGAR IS 6051 04:27:18,840 --> 04:27:21,160 IN THE STRUCTURE IT FOUND. 6052 04:27:21,160 --> 04:27:24,120 PROVIDES LINK TO PDB ENTRY, A 6053 04:27:24,120 --> 04:27:25,760 LINK TO THE VIEWER OF THE 6054 04:27:25,760 --> 04:27:28,520 STRUCTURE IF YOU WANT TO SPIN IT 6055 04:27:28,520 --> 04:27:31,840 AROUND AND DROP DOWN TO DISPLAY 6056 04:27:31,840 --> 04:27:37,080 MORE INFO. WHEN I TALK UNIFORM 6057 04:27:37,080 --> 04:27:38,880 NOMENCLATURE LET'S LOOK AT THESE 6058 04:27:38,880 --> 04:27:40,120 EXAMPLES. LOOK AT THE ENDS OF 6059 04:27:40,120 --> 04:27:42,040 THEM, ONE IS ATTACHED TO A 6060 04:27:42,040 --> 04:27:45,200 GLUTENATE. SUGAR ATTACHED TO 6061 04:27:45,200 --> 04:27:46,720 GLUTAMATE, NOT SUPER COMMON BUT 6062 04:27:46,720 --> 04:27:48,480 THERE IT IS. THERE IS A SUGAR 6063 04:27:48,480 --> 04:27:50,120 WITH OH ON THE END. THAT IS NOT 6064 04:27:50,120 --> 04:27:51,040 ATTACHED TO ANYTHING. CLEARLY 6065 04:27:51,040 --> 04:27:54,400 THAT IS A LIGANDS. AND THERE IS 6066 04:27:54,400 --> 04:27:57,120 SUGAR ATTACHED TO ASPEAR GENE SO 6067 04:27:57,120 --> 04:27:58,280 THAT IS A IMMUNOGLYCAN. 6068 04:27:58,280 --> 04:28:00,040 SO THIS REPRESENTS THE THREE 6069 04:28:00,040 --> 04:28:01,520 TYPES OF THINGS TO PULL OUT OF 6070 04:28:01,520 --> 04:28:06,040 THE DATA. IF YOU CLICK MORE 6071 04:28:06,040 --> 04:28:07,680 BUTTON YOU GET A COUPLE OF BITS 6072 04:28:07,680 --> 04:28:08,920 OF INFORMATION THAT I THINK ARE 6073 04:28:08,920 --> 04:28:11,960 QUITE USEFUL. IN TERMS OF THE 6074 04:28:11,960 --> 04:28:13,960 QUALITY OF STRUCTURE, 6075 04:28:13,960 --> 04:28:15,120 CRYSTALLOGRAPHERS OFTEN REFER TO 6076 04:28:15,120 --> 04:28:16,760 B FACTOR WHICH IS THE 6077 04:28:16,760 --> 04:28:18,320 UNCERTAINTY OF THE ATOMIC 6078 04:28:18,320 --> 04:28:21,560 POSITION AND THEIR STRUCTURE. 6079 04:28:21,560 --> 04:28:22,760 AND THEY WILL REPORT SOMETIMES 6080 04:28:22,760 --> 04:28:25,760 THE MEAN B FACTOR OF THE PROTEIN 6081 04:28:25,760 --> 04:28:28,840 BUT NOT OF ANYTHING ELSE. 6082 04:28:28,840 --> 04:28:31,080 BECAUSE THESE SUGARS ARE POORLY 6083 04:28:31,080 --> 04:28:32,800 RESOLVED WE THOUGHT LET'S REPORT 6084 04:28:32,800 --> 04:28:34,160 THE MEAN B FACTOR OF THE 6085 04:28:34,160 --> 04:28:36,080 OLIGOSACCHARIDE. SO NOW WHEN 6086 04:28:36,080 --> 04:28:39,720 YOU DO THIS, AND THINK IS THIS 6087 04:28:39,720 --> 04:28:40,880 STRUCTURE REASONABLE, THIS CAME 6088 04:28:40,880 --> 04:28:42,520 OUT RECENT WILL I IN A 6089 04:28:42,520 --> 04:28:45,840 PUBLICATION COMING OUT WITH BOB 6090 04:28:45,840 --> 04:28:47,480 WHERE E H SAID FUCOSE IS IN A 6091 04:28:47,480 --> 04:28:49,880 DIFFERENT ORIENTATION THAN YOUR 6092 04:28:49,880 --> 04:28:54,320 MODEL SAYS, THE MEAN B FACTOR IS 6093 04:28:54,320 --> 04:28:56,600 SOMETHING 65 OVER 95. IT IS 6094 04:28:56,600 --> 04:28:59,960 FANTASY LAND. KNOWING THIS YOU 6095 04:28:59,960 --> 04:29:01,280 CAN HAVE QUANTITATIVE ASSESSMENT 6096 04:29:01,280 --> 04:29:03,200 OF THE QUALITY OF THE 6097 04:29:03,200 --> 04:29:06,080 COORDINATES FOR THE SUGAR. THE 6098 04:29:06,080 --> 04:29:07,360 OTHER THING YOU GET IS SOMETHING 6099 04:29:07,360 --> 04:29:09,160 ON THE BOTTOM, IF THERE IS 6100 04:29:09,160 --> 04:29:11,760 ANYTHING THAT DISAGREES BETWEEN 6101 04:29:11,760 --> 04:29:13,840 PDB NOMENCLATURE OR DATA, AND 6102 04:29:13,840 --> 04:29:17,600 WHAT WE THINK IT IS. IN THIS 6103 04:29:17,600 --> 04:29:18,840 CASE IF YOU LOOK AT THE 6104 04:29:18,840 --> 04:29:21,600 STRUCTURE IT IS MURNAC. 6105 04:29:21,600 --> 04:29:23,080 CURRENTLY IN OUR NAMING SYSTEM 6106 04:29:23,080 --> 04:29:25,520 WE DON'T HAVE ONE SO WE CALL 6107 04:29:25,520 --> 04:29:29,280 THIS A WEIRD GLCNAC WITH AN RO 3 6108 04:29:29,280 --> 04:29:31,400 MODIFICATION. SO THIS IS A 6109 04:29:31,400 --> 04:29:32,640 MISMATCH BUT GIVES YOU HIGHLIGHT 6110 04:29:32,640 --> 04:29:36,920 SOMETIMES IT IS US THAT DOESN'T 6111 04:29:36,920 --> 04:29:38,080 UNDERSTAND, SOMETIMES THE PDB 6112 04:29:38,080 --> 04:29:43,640 THAT HAS INCORRECT. IF YOU THEN 6113 04:29:43,640 --> 04:29:45,640 CLICK THIS AND WISH I COULD LYE, 6114 04:29:45,640 --> 04:29:46,960 IF YOU CAN SEE ICON 6115 04:29:46,960 --> 04:29:49,840 REPRESENTATION OF THE SUGAR AND 6116 04:29:49,840 --> 04:29:52,600 YOU CAN SPIN AROUND AND LOOK 6117 04:29:52,600 --> 04:29:57,480 YOURSELF AT THE 3-D STRUCTURE. 6118 04:29:57,480 --> 04:30:01,360 IN TERMS OF PROBETY WE CALLED 6119 04:30:01,360 --> 04:30:07,360 THE TOOLS GLY FINDER AND 6120 04:30:07,360 --> 04:30:10,720 GLYPROBITY. THEY MAY SPLIT OUT, 6121 04:30:10,720 --> 04:30:12,920 GLYPROBITY IS NAMED AFTER A TOOL 6122 04:30:12,920 --> 04:30:15,400 THAT CRYSTALLOGRAPHERS USE FOR 6123 04:30:15,400 --> 04:30:16,920 LOOKING AT CONFIDENCE OF THE 6124 04:30:16,920 --> 04:30:19,240 COORDINATES OF THE PROTEIN. 6125 04:30:19,240 --> 04:30:22,320 THERE IS SOME NEED FOR THIS FOR 6126 04:30:22,320 --> 04:30:23,640 SUGARS AS WELL. ONE THING WE 6127 04:30:23,640 --> 04:30:25,880 DEVELOPED THAT CAME OUT THIS, 6128 04:30:25,880 --> 04:30:27,680 BECAUSE WE DO THEORETICAL MODELK 6129 04:30:27,680 --> 04:30:29,560 OF MOLECULAR STRUCTURE, WE ARE 6130 04:30:29,560 --> 04:30:30,840 OFTEN LOOKING AT ENERGY OF 6131 04:30:30,840 --> 04:30:33,720 DIFFERENT SHAPES. CARBOHYDRATES 6132 04:30:33,720 --> 04:30:39,520 HAVE WELL DEFINED CONFIRMATIONAL 6133 04:30:39,520 --> 04:30:42,200 PREFERENCE AROUND GLYCOSIDIC 6134 04:30:42,200 --> 04:30:44,160 LINKAGE, WE CAN BUILD 3-D MODEL 6135 04:30:44,160 --> 04:30:45,120 OF SUGARS BECAUSE WE KNOW HOW 6136 04:30:45,120 --> 04:30:46,680 THE LINKAGES BEHAVE, YOU CAN'T 6137 04:30:46,680 --> 04:30:48,000 DO IT WITH PROTEINS BECAUSE THEY 6138 04:30:48,000 --> 04:30:51,080 ARE TOO FLEXIBLE. ALPHA FOLD 6139 04:30:51,080 --> 04:30:54,200 SOLVE PROTEIN MODELING PROBLEM, 6140 04:30:54,200 --> 04:30:57,400 THIS RECENTLY, CARBOHYDRATE 6141 04:30:57,400 --> 04:30:59,960 MODELING PROBLEM SOLVED FOR A 6142 04:30:59,960 --> 04:31:01,720 LONG TIME BECAUSE WE UNDERSTAND 6143 04:31:01,720 --> 04:31:03,560 LINKAGES BEHAVE. WHAT THAT LETS 6144 04:31:03,560 --> 04:31:05,720 US DO IS CREATE IMAGE LIKE THIS 6145 04:31:05,720 --> 04:31:10,360 AKIN TO A RAMISHANDRIN PLOT FOR 6146 04:31:10,360 --> 04:31:13,240 PROTEIN AND BLACK DOTS WHERE 6147 04:31:13,240 --> 04:31:14,040 WHERE THE CRYSTAL STRUCTURES 6148 04:31:14,040 --> 04:31:18,200 HAVE THE THIAN ANGLES, THE 6149 04:31:18,200 --> 04:31:20,720 SHAPES. COLORS WE INDEPENDENTLY 6150 04:31:20,720 --> 04:31:21,360 PREDICT ENERGY OF THESE 6151 04:31:21,360 --> 04:31:23,760 STRUCTURE TO BE. SO YOU CAN SEE 6152 04:31:23,760 --> 04:31:25,760 THIS IS A VERY INTERESTING BASED 6153 04:31:25,760 --> 04:31:27,960 ON QUANTUM CALCULATION OF ENERGY 6154 04:31:27,960 --> 04:31:29,640 SURFACE, AND WHAT IT SHOWS IS 6155 04:31:29,640 --> 04:31:31,080 THERE IS -- THE BLUE IS STABLE 6156 04:31:31,080 --> 04:31:33,400 STUFF AND THAT IS WHERE MOST 6157 04:31:33,400 --> 04:31:36,160 STRUCTURE ARE, IT PROVIDES VERY 6158 04:31:36,160 --> 04:31:37,280 OBJECTIVE INDEPENDENT 6159 04:31:37,280 --> 04:31:38,600 ASSESSMENT, IT ALSO LETS YOU 6160 04:31:38,600 --> 04:31:40,760 FIND WHAT ARE CALLED OUTLIERS. 6161 04:31:40,760 --> 04:31:42,720 THIS IS AN INTERESTING TERM, 6162 04:31:42,720 --> 04:31:46,920 DOESN 'T MEAN MISTAKE. IT MEANS 6163 04:31:46,920 --> 04:31:47,760 STATISTICALLY NOT COMMON. THIS 6164 04:31:47,760 --> 04:31:49,320 COULD BE MISTAKE AND PROBABLY IS 6165 04:31:49,320 --> 04:31:52,480 WITH SUPER HIGH ENERGY, PROBABLY 6166 04:31:52,480 --> 04:31:53,520 INCORRECT STRUCTURE, YOU CAN 6167 04:31:53,520 --> 04:31:55,760 FLIP A SUGAR 180 DEGREES IN 6168 04:31:55,760 --> 04:31:57,320 ELECTRON DENSITY AND NOT KNOW 6169 04:31:57,320 --> 04:31:58,240 THE DIFFERENCE UNLESS YOU ARE 6170 04:31:58,240 --> 04:32:04,360 CAREFUL. BUT COULD BE ENZYME HAS 6171 04:32:04,360 --> 04:32:06,800 DISTORTED IT. UNLESS YOU FIND 6172 04:32:06,800 --> 04:32:10,960 THINGS LIKE THIS. WHAT WE ARE 6173 04:32:10,960 --> 04:32:13,840 STRUGGLING WITH, EVOLVING TO DO, 6174 04:32:13,840 --> 04:32:16,360 WHEN IS A CARBOHYDRATE NOT A 6175 04:32:16,360 --> 04:32:18,160 CARBOHYDRATE? THERE ARE LOTS OF 6176 04:32:18,160 --> 04:32:19,160 MODIFICATIONS AND DERIVATIVES. 6177 04:32:19,160 --> 04:32:22,000 SO THESE ARE THE THINGS WHERE AT 6178 04:32:22,000 --> 04:32:23,720 MOMENT WE CALL THEM R GROUPS AND 6179 04:32:23,720 --> 04:32:28,680 WE WILL LIST THE C 612OH OF THE 6180 04:32:28,680 --> 04:32:30,640 R GROWN BUT GIVE IT A NAME 6181 04:32:30,640 --> 04:32:31,960 BECAUSE IT IS JUST NOT WORTH THE 6182 04:32:31,960 --> 04:32:33,760 EFFORT OF TRYING TO NAME THESE R 6183 04:32:33,760 --> 04:32:35,120 GROUPS. BUT IT DOES DETECT THEM 6184 04:32:35,120 --> 04:32:37,320 AND THIS IS A VERY POWERFUL 6185 04:32:37,320 --> 04:32:39,120 THING THAT WHEN WE FIRST STARTED 6186 04:32:39,120 --> 04:32:40,560 TO THIS PROJECT WE WEREN'T 6187 04:32:40,560 --> 04:32:42,400 THINKING THIS, WE WERE THINKING 6188 04:32:42,400 --> 04:32:44,120 OF LOOKING FOR CAR BOAR HYDRATES 6189 04:32:44,120 --> 04:32:45,360 BUT WE GOT SUPPLEMENT THAT 6190 04:32:45,360 --> 04:32:47,160 HELPED US GET THE MODIFICATIONS 6191 04:32:47,160 --> 04:32:50,560 AS WELL. WHAT DID WE LEARN FROM 6192 04:32:50,560 --> 04:32:54,920 THIS? JUST LOOKING THERE IS 54, 6193 04:32:54,920 --> 04:32:58,880 55% SUGARS IN THE PDB ARE N 6194 04:32:58,880 --> 04:33:03,400 LINKED SUGARS. SO SUGARS FAR 6195 04:33:03,400 --> 04:33:04,400 FEWER. I DON'T KNOW IF THAT IS 6196 04:33:04,400 --> 04:33:06,400 BECAUSE THEY ARE NOT LOOKED FOR, 6197 04:33:06,400 --> 04:33:09,360 WHICH IS DEFINITELY POSSIBLE. OR 6198 04:33:09,360 --> 04:33:13,000 JUST NOT THERE. CEILINGS ARE 6199 04:33:13,000 --> 04:33:14,920 FURTHER -- C LUNGE IS FURTHER 6200 04:33:14,920 --> 04:33:17,440 UNLIKELY. WHAT CAN YOU DO A 6201 04:33:17,440 --> 04:33:20,800 LITTLE FACTOID TO AMUSE FRIENDS, 6202 04:33:20,800 --> 04:33:23,000 52,000 IMMUNOGLYCANS OUT OF 6203 04:33:23,000 --> 04:33:24,400 7,000 SOME ODD STRUCTURE 6204 04:33:24,400 --> 04:33:26,640 CONTAINING N LINK GLYCANS THAT 6205 04:33:26,640 --> 04:33:29,120 MEANS SOME N LINK GLYCANS ON 6206 04:33:29,120 --> 04:33:30,960 AVERAGE PER GLYCOPROTEIN. WHO 6207 04:33:30,960 --> 04:33:32,520 KNEW THIS? NOBODY. WE COULDN'T. 6208 04:33:32,520 --> 04:33:33,280 DOES IT MATTER I DON'T KNOW BUT 6209 04:33:33,280 --> 04:33:36,640 IT IS A FACT. THAT IS ALL I WANT 6210 04:33:36,640 --> 04:33:39,440 TO TALK ABOUT. I WILL THANK THE 6211 04:33:39,440 --> 04:33:42,320 PEOPLE INVOLVED. THIS IS THE GLY 6212 04:33:42,320 --> 04:33:46,400 FINDER WORK UNDERTALKEN BY A 6213 04:33:46,400 --> 04:33:48,000 TALENTED GRAD STUDENT DAVID 6214 04:33:48,000 --> 04:33:50,480 MONTGOMERY. I WOULD LIKE TO 6215 04:33:50,480 --> 04:33:51,920 ACKNOWLEDGE THE NIH AND 6216 04:33:51,920 --> 04:33:53,160 UNIVERSITY OF GEORGIA AND ALL 6217 04:33:53,160 --> 04:33:54,600 THE TEAM, YOU GUYS FOR THE TIME. 6218 04:33:54,600 --> 04:34:04,160 THANK YOU. 6219 04:34:04,160 --> 04:34:07,560 >> NEXT SPEAKER IS DR. RILEY AND 6220 04:34:07,560 --> 04:34:09,560 HE TALK ABOUT MAKING 6221 04:34:09,560 --> 04:34:15,160 GLYCOPROTEOMICS MORE ACCESSIBLE. 6222 04:34:15,160 --> 04:34:16,480 >> THANK YOU, MICROPHONE WORKS 6223 04:34:16,480 --> 04:34:22,680 TOO. THANK YOU ALL FOR ALLOWING 6224 04:34:22,680 --> 04:34:25,920 ME TO GIVE THIS PRESENTATION, 6225 04:34:25,920 --> 04:34:28,320 HERE REPRESENTING BERTOZZI GROUP 6226 04:34:28,320 --> 04:34:29,480 AT STANFORD, THANK YOU FOR 6227 04:34:29,480 --> 04:34:30,880 SENDING ME TO PRESENT TO Y'ALL 6228 04:34:30,880 --> 04:34:32,880 AND TO ORGANIZE ORS FOR AGREEING 6229 04:34:32,880 --> 04:34:34,200 TO LET ME DO THIS. I'M 6230 04:34:34,200 --> 04:34:36,080 PRESENTING ON THIS UO 1 SHARED 6231 04:34:36,080 --> 04:34:37,920 BETWEEN CAROLYN AND SHARON AT 6232 04:34:37,920 --> 04:34:39,680 STANFORD. AND WE ARE EXCITED TO 6233 04:34:39,680 --> 04:34:40,760 TALK ABOUT BOTH WHAT WE HAVE 6234 04:34:40,760 --> 04:34:42,120 DONE IN THE FUNDING PERIOD AND 6235 04:34:42,120 --> 04:34:45,680 ALSO WHAT THIS WORK IS GENERATED 6236 04:34:45,680 --> 04:34:46,480 AS YOUR NEXT IDEAS AND WORKING 6237 04:34:46,480 --> 04:34:53,040 ON MOST RECENT FEW YEARS. 6238 04:34:53,040 --> 04:34:54,440 CONFLICT OF INTEREST, CAROLYN 6239 04:34:54,440 --> 04:34:55,720 HAS A FEW COMPANIES SHE'S PART 6240 04:34:55,720 --> 04:35:06,480 OF SO MENTION THAT. I HOPE MY 6241 04:35:06,480 --> 04:35:07,160 STORY IS ONE SMALL 6242 04:35:07,160 --> 04:35:08,160 REPRESENTATION OF SUCCESS OF 6243 04:35:08,160 --> 04:35:09,200 WHAT THE GLYCOSCIENCE COMMON 6244 04:35:09,200 --> 04:35:11,080 FUND HAS BEEN. THAT'S MY THIRD 6245 04:35:11,080 --> 04:35:12,880 IN PERSON COMMON FUND I WENT TO 6246 04:35:12,880 --> 04:35:16,360 BECAUSE IN GRADUATE SCHOOL AS 6247 04:35:16,360 --> 04:35:17,680 MEMBER PROFESSOR JOSH COUNSEL'S 6248 04:35:17,680 --> 04:35:20,560 GROUP UW MADISON MADISON. WE GOT AN R21 6249 04:35:20,560 --> 04:35:21,920 TO WORK ON HARDWARE DEVELOPMENT 6250 04:35:21,920 --> 04:35:23,520 HOW TO ANALYZE GLYCOPEPTIDES 6251 04:35:23,520 --> 04:35:25,680 USING INNOVATIVE ELECTRON BASED 6252 04:35:25,680 --> 04:35:26,800 AUGMENTATION WE WORKED ON IN 6253 04:35:26,800 --> 04:35:30,200 MASS SPEC. IN THOSE MEETINGS, 6254 04:35:30,200 --> 04:35:31,360 FIRST TIME I WAS IN THE ROOM 6255 04:35:31,360 --> 04:35:33,160 WITH CAROLYN SHE ASKED QUESTION 6256 04:35:33,160 --> 04:35:35,160 ABOUT PRESENTATION AND LITTLE 6257 04:35:35,160 --> 04:35:38,160 DID I KNOW IN 2016 THAT HAPPENED 6258 04:35:38,160 --> 04:35:40,160 AND CONTACT OR TWO LATER DO YOU 6259 04:35:40,160 --> 04:35:42,360 HAVE POST DOC OPENINGS, HERE I 6260 04:35:42,360 --> 04:35:43,960 AM SIX YEARS AFTER FACT. IT IS 6261 04:35:43,960 --> 04:35:45,800 EXCITING TO TALK ABOUT THE UO 1 6262 04:35:45,800 --> 04:35:47,560 WITH HER. THIS FUNDING PERIOD 6263 04:35:47,560 --> 04:35:49,800 ENDED A FEW YEARS AGO. WHAT IS 6264 04:35:49,800 --> 04:35:50,960 INTERESTING ABOUT THIS IS WORK 6265 04:35:50,960 --> 04:35:53,800 THAT CAME OUT OF UO 1 GRANT WAS 6266 04:35:53,800 --> 04:35:56,320 LOTS OF SPEARHEADED BY KRISTINA 6267 04:35:56,320 --> 04:35:59,880 WOO WHO TALK ABOUT HER WORK IN 6268 04:35:59,880 --> 04:36:02,120 FRONT OF YOU AND YOU. WE WANT TO 6269 04:36:02,120 --> 04:36:03,920 HIGHLIGHT HOW THAT TRANSLATED TO 6270 04:36:03,920 --> 04:36:04,720 PRODUCT AVAILABLE IN COMMUNITY 6271 04:36:04,720 --> 04:36:07,080 AND WAYS WE ARE STILL TRYING TO 6272 04:36:07,080 --> 04:36:10,360 MAKE GLYCOPROTEOMICS AND MASS 6273 04:36:10,360 --> 04:36:11,400 SPECTROMETRY ACCESSIBLE TO THOSE 6274 04:36:11,400 --> 04:36:15,600 WITH GLYCOPROTEINS. AS WE TALKED 6275 04:36:15,600 --> 04:36:17,560 ABOUT DIVERSE GLYCOBIOLOGY 6276 04:36:17,560 --> 04:36:18,960 DIVERSE SETS OF TOOLS AND 6277 04:36:18,960 --> 04:36:21,160 CAROLYN APPROACH TO THIS IS TO 6278 04:36:21,160 --> 04:36:22,160 USE THESE METS BOLLICLY 6279 04:36:22,160 --> 04:36:23,640 INCORPORATED SUGARS AND 6280 04:36:23,640 --> 04:36:25,640 NUCLEOTIDE SUGARS THROUGHOUT 6281 04:36:25,640 --> 04:36:26,960 THESE PRESENTATIONS AND 6282 04:36:26,960 --> 04:36:28,560 INCORPORATE INTO GLYCAN TO DO 6283 04:36:28,560 --> 04:36:36,280 INTERESTING COOL CHEMISTRY. 6284 04:36:36,280 --> 04:36:37,760 GLYCOTON JEW GATES. YOU CAN 6285 04:36:37,760 --> 04:36:39,120 TARGET GLYCANS DEPENDING ON TYPE 6286 04:36:39,120 --> 04:36:40,520 OF METABOLIC SUGAR YOU 6287 04:36:40,520 --> 04:36:47,680 INCORPORATE THAT. WITH Z FOCUS 6288 04:36:47,680 --> 04:36:49,960 ON THIS CYTOTAG PLATFORM TO USE 6289 04:36:49,960 --> 04:36:51,080 ANY SUGAR YOU CAN IMAGINE TO 6290 04:36:51,080 --> 04:36:52,800 HAVE METABOLIC REPORTER THAT IS 6291 04:36:52,800 --> 04:36:54,360 FUNCTIONALLIZE.MENT SO YOU CAN 6292 04:36:54,360 --> 04:36:56,840 INCORPORATE THIS SUGAR INTO THE 6293 04:36:56,840 --> 04:36:57,600 GLYCOCALIX OR INTO 6294 04:36:57,600 --> 04:37:00,040 O-GlcNac PROTEINS AS WELL 6295 04:37:00,040 --> 04:37:01,160 DEPENDING WHICH MONOSACCHARIDES 6296 04:37:01,160 --> 04:37:02,360 YOU USE AND NEEDS TO BE 6297 04:37:02,360 --> 04:37:06,200 PRESENTED ON CELL SURFACE OR 6298 04:37:06,200 --> 04:37:06,760 O-GlcNacATEED 6299 04:37:06,760 --> 04:37:07,800 INTRACELLULAR PROTEINS TOO. 6300 04:37:07,800 --> 04:37:11,960 THIS IS A HANDLE ON 6301 04:37:11,960 --> 04:37:12,720 GLYCOCONJUGATES AS WE HAVE SEEN 6302 04:37:12,720 --> 04:37:15,560 THROUGHOUT PRESENTATIONS TODAY. 6303 04:37:15,560 --> 04:37:17,720 WITH ISOTYPE PROTEIN ONCE YOU 6304 04:37:17,720 --> 04:37:19,800 HAVE THE LABELED PROTEIN, THIS 6305 04:37:19,800 --> 04:37:21,840 GLYCOCONJUGATE HOW YOU CAPTURE 6306 04:37:21,840 --> 04:37:24,040 AND DO MASS SPEC TO UNDERSTAND 6307 04:37:24,040 --> 04:37:25,560 NOT THAT IT WAS GLYCOSYLATED BUT 6308 04:37:25,560 --> 04:37:27,360 WHAT IS THE GLYCOSIDE AND WHAT 6309 04:37:27,360 --> 04:37:29,280 GLYCAN WAS THERE SO LOOKING AT 6310 04:37:29,280 --> 04:37:31,280 INTACT GLYCOPEPTIDES. AND THEN 6311 04:37:31,280 --> 04:37:32,600 IF YOU HAVE A CHEMICAL 6312 04:37:32,600 --> 04:37:33,960 REPRESENTATION HOW THIS MIGHT 6313 04:37:33,960 --> 04:37:35,400 WORK YOU CAN DEVELOP REAGENT, 6314 04:37:35,400 --> 04:37:37,920 THIS IS ISOTAG REAGENT BY 6315 04:37:37,920 --> 04:37:40,360 KRISTINA AND THE BERTOZZI LAB, 6316 04:37:40,360 --> 04:37:42,960 TO BE ABLE TO CLICK THIS ON WITH 6317 04:37:42,960 --> 04:37:45,760 CHEMISTRY TO OUR LABEL 6318 04:37:45,760 --> 04:37:48,600 GLYCOPROTEIN DO ON B DIGESTION 6319 04:37:48,600 --> 04:37:52,800 SOME WAY TO PURIFY LABELED 6320 04:37:52,800 --> 04:37:53,920 GLYCOPROTEINS AND LINK THROUGH 6321 04:37:53,920 --> 04:37:56,960 ASSAY LINKERS, DIFFERENT WAYS TO 6322 04:37:56,960 --> 04:37:58,160 RELEASE GLYCOCONJUGATE TO BE 6323 04:37:58,160 --> 04:38:01,080 ABLE TO ANALYZE. SO WHAT THESE 6324 04:38:01,080 --> 04:38:03,160 TAGS LOOK LIKE THESE THINGS HAVE 6325 04:38:03,160 --> 04:38:05,120 MULTI-FUNCTIONAL PARTS TO THEM. 6326 04:38:05,120 --> 04:38:07,480 WE HAVE THE BIOTIN GROUP HERE 6327 04:38:07,480 --> 04:38:09,880 USED FOR BASED ENRICHMENT, THE 6328 04:38:09,880 --> 04:38:11,400 LINKER THAT GETS TO CLEAVAGE 6329 04:38:11,400 --> 04:38:14,480 SIDE WHICH IS WHERE YOU RELEASE 6330 04:38:14,480 --> 04:38:16,360 THIS MOLECULE FROM ENRICHMENT 6331 04:38:16,360 --> 04:38:17,960 AND YOU HAVE ISOTOPE RECODING OR 6332 04:38:17,960 --> 04:38:20,360 RECORDING. AND SO HERE YOU 6333 04:38:20,360 --> 04:38:24,960 INTRODUCE AN ISOTOPIC PATTERN TO 6334 04:38:24,960 --> 04:38:26,680 DIFFERENCE FROM PATTERN PRESENT 6335 04:38:26,680 --> 04:38:27,440 IN GLYCOPROTENSE OR GLYCOPEP 6336 04:38:27,440 --> 04:38:29,200 SIDES, THIS IN YOUR MASS 6337 04:38:29,200 --> 04:38:30,520 SPECTROMETER IS A DIFFERENT 6338 04:38:30,520 --> 04:38:33,920 PATTERN. THIS IS A NOW ENRICHED 6339 04:38:33,920 --> 04:38:35,480 GLYCOCONJUGATE LIKE A PEPTIDE 6340 04:38:35,480 --> 04:38:36,600 PROTEIN OF SYSTEM BILL THAT I 6341 04:38:36,600 --> 04:38:38,080 WANT TO STUDY. WITH CLICKABLE 6342 04:38:38,080 --> 04:38:40,360 SIDE YOU INTRODUCE CHEMICAL 6343 04:38:40,360 --> 04:38:42,560 FUNCTIONALITIES TO ATTACH TO THE 6344 04:38:42,560 --> 04:38:43,960 METABOLIC SUGAR YOU HAVE 6345 04:38:43,960 --> 04:38:45,840 INCORPORATE WITH THE RECIPROCAL 6346 04:38:45,840 --> 04:38:51,960 FUNCTIONALITY ON I ALABAMATY ON IT. SO 6347 04:38:51,960 --> 04:38:55,840 THIS IN-- ISOTAG INCORPORATES 6348 04:38:55,840 --> 04:38:57,360 THE IDEA, THE SECOND PART IS HOW 6349 04:38:57,360 --> 04:38:59,160 THE ANALYZE WITH MASS 6350 04:38:59,160 --> 04:39:00,160 SPECTROMETRY, THAT WAS THE IDEA 6351 04:39:00,160 --> 04:39:03,760 OF UO 1 TO BRING THIS IDEA TO 6352 04:39:03,760 --> 04:39:06,760 THE MASSES SO PEOPLE CAN DO 6353 04:39:06,760 --> 04:39:08,160 GLYCOPEPTIDE MEASUREMENTS. SO 6354 04:39:08,160 --> 04:39:09,960 THE IDEA HERE WITH THE ISOTOPE 6355 04:39:09,960 --> 04:39:11,880 RECODING IS YOU HAVE THIS IDEA 6356 04:39:11,880 --> 04:39:14,360 THAT WE NOW HAVE A ISOTOPE THAT 6357 04:39:14,360 --> 04:39:15,480 LOOKS DIFFERENCE THAN NORMAL 6358 04:39:15,480 --> 04:39:17,480 PEPTIDE WOULD. WE CAN TELL MASS 6359 04:39:17,480 --> 04:39:19,080 SPECTROMETER GO AFTER THAT AND 6360 04:39:19,080 --> 04:39:20,760 CLEAVE -- ANALYZE THAT OVER 6361 04:39:20,760 --> 04:39:21,960 PREFERENCE OF ANYTHING ELSE AND 6362 04:39:21,960 --> 04:39:24,040 THAT GIVES YOU DATA THAT CAN SAY 6363 04:39:24,040 --> 04:39:25,360 THESE SPECTRA SHOULD BE 6364 04:39:25,360 --> 04:39:29,520 GLYCOPEPTIDES. SO WITH THIS 6365 04:39:29,520 --> 04:39:31,240 IDEA WE CAN ANALYZE SPECIALIZED 6366 04:39:31,240 --> 04:39:32,920 SOFTWARE, WORK WITH PROTEIN 6367 04:39:32,920 --> 04:39:35,480 METRICS TO -- JENNIFER KOHLER 6368 04:39:35,480 --> 04:39:36,440 MENTIONED WORKING WITH THEM AS 6369 04:39:36,440 --> 04:39:38,400 PART OF FUNDING MECHANISM, SAME 6370 04:39:38,400 --> 04:39:42,560 WITH BERTOZZI LAB TO BE ABLE TO 6371 04:39:42,560 --> 04:39:43,600 CHARACTERIZE SPECIFIC 6372 04:39:43,600 --> 04:39:46,440 GLYCOPEPTIDES THAT WERE ENRICHED 6373 04:39:46,440 --> 04:39:48,360 THROUGH THE ISOTAG PLATFORM. I 6374 04:39:48,360 --> 04:39:49,960 WILL MENTION TOO THE GOAL OF THE 6375 04:39:49,960 --> 04:39:51,760 UO 1 WAS TO GENERATE A 6376 04:39:51,760 --> 04:39:52,720 COMMERCIALLY AVAILABLE PRODUCT 6377 04:39:52,720 --> 04:39:54,560 PEOPLE CAN DO THIS SO THIS HAS 6378 04:39:54,560 --> 04:39:56,440 COME TO FRUITION THROUGH CLICK 6379 04:39:56,440 --> 04:39:57,560 CHEMISTRY TOOLS, YOU CAN SEE 6380 04:39:57,560 --> 04:39:59,560 THERE IS MULTIPLE OPTIONS YOU 6381 04:39:59,560 --> 04:40:01,000 CAN PURCHASE AND DO THIS YOUR 6382 04:40:01,000 --> 04:40:02,560 LAB FOR RELATIVELY AFFORDABLE 6383 04:40:02,560 --> 04:40:04,800 PRICES. THE PRICE OF ANTIBODY 6384 04:40:04,800 --> 04:40:06,680 YOU MIGHT NEED TO DO FOR 6385 04:40:06,680 --> 04:40:08,040 EXPERIMENT. YOU CAN BUY THE 6386 04:40:08,040 --> 04:40:09,840 ENTIRE KIT OR YOU CAN BUY JUST 6387 04:40:09,840 --> 04:40:12,840 THE LINKER ENRICHMENT SET THAT 6388 04:40:12,840 --> 04:40:14,120 TOOL SO IF YOU HAVE YOUR OWN WAY 6389 04:40:14,120 --> 04:40:17,560 TO SYNTHESIZE YOUR NUCLEOTIDE 6390 04:40:17,560 --> 04:40:18,120 SUGARS, METS BOLLIC 6391 04:40:18,120 --> 04:40:18,760 INCORPORATION MECHANISMS YOU CAN 6392 04:40:18,760 --> 04:40:21,480 DO THAT AND STILL USE THIS IDEA 6393 04:40:21,480 --> 04:40:23,720 OF THE ISOTAG PLATFORM SO YOU 6394 04:40:23,720 --> 04:40:25,080 CAN SEE ALL THIS AVAILABLE AT 6395 04:40:25,080 --> 04:40:27,000 CLICK CHEMISTRY TOOLS, THIS 6396 04:40:27,000 --> 04:40:29,440 REALLY WAS ENTIRE PATH OF THIS 6397 04:40:29,440 --> 04:40:30,400 PROJECT US TO BRING TO COMMUNITY 6398 04:40:30,400 --> 04:40:31,400 AND NOW EXCITED THAT IT IS 6399 04:40:31,400 --> 04:40:36,240 AVAILABLE THAT WAY. KAREN AND 6400 04:40:36,240 --> 04:40:38,240 SHARON WERE PIs, KRISTINA DID 6401 04:40:38,240 --> 04:40:40,000 WORK TO GENERATE THIS AND THESE 6402 04:40:40,000 --> 04:40:41,160 ARE SELECTIVE REFERENCE BUS THEY 6403 04:40:41,160 --> 04:40:42,760 HAVE AN ENTIRE BODY OF WORK 6404 04:40:42,760 --> 04:40:45,640 TALKING ABOUT HOW ISOTAG IS USED 6405 04:40:45,640 --> 04:40:51,560 FOR N GLYCANS O GALNAC AND 6406 04:40:51,560 --> 04:40:52,240 O-GlcNac GLYCOPROTENSE SO 6407 04:40:52,240 --> 04:40:54,120 DIFFERENT WAYS TO USE THE 6408 04:40:54,120 --> 04:40:55,360 TECHNOLOGY AND WE CAN SPEND THE 6409 04:40:55,360 --> 04:40:56,840 ENTIRE REST OF THE AFTERNOON 6410 04:40:56,840 --> 04:40:58,200 TALKING ABOUT DIFFERENT AWESOME 6411 04:40:58,200 --> 04:41:00,560 DATA THEY GENERATED THAT WAY. 6412 04:41:00,560 --> 04:41:02,760 BUT INSTEAD WE WANT TO PIVOT, 6413 04:41:02,760 --> 04:41:03,840 CAROLYN SAID YOU SHOULD TALK 6414 04:41:03,840 --> 04:41:05,200 ABOUT WORK THAT'S GROWN FROM 6415 04:41:05,200 --> 04:41:06,560 THIS, WHAT WE ARE DOING NOW WITH 6416 04:41:06,560 --> 04:41:08,760 IDEAS FROM THE UO 1 THROUGH THIS 6417 04:41:08,760 --> 04:41:11,240 MECHANISM. SO WANTS TO POINT OUT 6418 04:41:11,240 --> 04:41:13,760 THIS IDEA OF WHAT IS MS BASED 6419 04:41:13,760 --> 04:41:14,560 GLYCOPROTEOMICS AND YOU HAVE 6420 04:41:14,560 --> 04:41:16,360 SEEN WORK FLOWS AND WHAT IT 6421 04:41:16,360 --> 04:41:17,720 BOILS DOWN TO IS MAIN COMPONENTS 6422 04:41:17,720 --> 04:41:19,360 YOU HAVE TO CONSIDER. WE CAN 6423 04:41:19,360 --> 04:41:20,760 GIVE SEMESTER LONG COURSES ABOUT 6424 04:41:20,760 --> 04:41:23,520 EACH OF THESE TOPICS. ISOTAG 6425 04:41:23,520 --> 04:41:25,160 TOUCHED ON SEVERAL POINTS. HOW 6426 04:41:25,160 --> 04:41:27,560 DO YOU INCORPORATE HANDLING YOUR 6427 04:41:27,560 --> 04:41:29,760 SAMPLES, ENRICH GLYCOPEPTIDES OR 6428 04:41:29,760 --> 04:41:30,360 PROTEINS. 6429 04:41:30,360 --> 04:41:32,840 HOW YOU ANALYZE WITH MASS 6430 04:41:32,840 --> 04:41:33,720 SPECTROMETER AND HOW TO SEARCH 6431 04:41:33,720 --> 04:41:35,160 IT. THERE'S PAIN POINTS 6432 04:41:35,160 --> 04:41:37,360 ESPECIALLY MORE CHALLENGING 6433 04:41:37,360 --> 04:41:38,040 GLYCOCONJUGATES IN FIELD. 6434 04:41:38,040 --> 04:41:39,840 SO WE WILL TALK ABOUT DATA 6435 04:41:39,840 --> 04:41:41,440 INFORMATICS BUT SAMPLE 6436 04:41:41,440 --> 04:41:42,800 PREPARATION CHEMICAL BIOLOGY 6437 04:41:42,800 --> 04:41:44,440 APPROACH LOOKING AT FOR SOME 6438 04:41:44,440 --> 04:41:46,440 CHALLENGING GLYCOPROTEINS. ONE 6439 04:41:46,440 --> 04:41:48,560 OF THE THINGS WE ARE MOST 6440 04:41:48,560 --> 04:41:50,720 INTERESTED IN STUDIES OF RECENT 6441 04:41:50,720 --> 04:41:53,560 YEARS ARE MUCINS. THESE ARE 6442 04:41:53,560 --> 04:41:55,360 MASSIVE OGLYCO PROTEINS WITH 6443 04:41:55,360 --> 04:41:59,760 DENSE REGIONS OF GLYCOSYLATION, 6444 04:41:59,760 --> 04:42:01,960 MOSTLY STUDIED IN DISEASES LIKE 6445 04:42:01,960 --> 04:42:04,800 CANCER, WE THINK OF MUC 1 6446 04:42:04,800 --> 04:42:07,160 REGULATED IN EPITHELIAL CANCER, 6447 04:42:07,160 --> 04:42:08,040 MUC 16 YOU HEARD ABOUT IN 6448 04:42:08,040 --> 04:42:09,160 EARLIER PRESENTATIONS AS MARKER 6449 04:42:09,160 --> 04:42:11,560 FOR OVARIAN CANCER. SO IT IS 6450 04:42:11,560 --> 04:42:13,240 REALLY INVOLVED IN 6451 04:42:13,240 --> 04:42:14,960 IMMUNOREGULATION AND EVASION, A 6452 04:42:14,960 --> 04:42:17,320 LOT OF HOST PATHOGEN 6453 04:42:17,320 --> 04:42:20,160 INTERACTIONS AND ALSO STRUCTURAL 6454 04:42:20,160 --> 04:42:20,760 MECHANOBIOLOGY, NOT JUST 6455 04:42:20,760 --> 04:42:21,880 CHEMISTRY OF LIGANDS BUT HOW 6456 04:42:21,880 --> 04:42:24,800 THEY REARRANGE CELL SURFACE AN 6457 04:42:24,800 --> 04:42:26,400 ALTER WAY SURFACE IS ORGANIZED. 6458 04:42:26,400 --> 04:42:28,320 SO LOT OF REASONS TO STUDY 6459 04:42:28,320 --> 04:42:31,560 MUCINS BUT THEY FRUSTRATE EVERY 6460 04:42:31,560 --> 04:42:35,360 STEP OF WHAT MASS SPEC BASED 6461 04:42:35,360 --> 04:42:36,000 GLYCOPROTEOMICS WORK FLOW LOOKS 6462 04:42:36,000 --> 04:42:37,360 LIKE SO THIS IS WHERE WE FOCUS 6463 04:42:37,360 --> 04:42:40,520 ON EFFORTS IN BERTOZZI LAB HOW 6464 04:42:40,520 --> 04:42:42,480 TO ADDRESS CHALLENGES WITH 6465 04:42:42,480 --> 04:42:43,480 DIFFERENT TOOLS WE CAN IMAGINE 6466 04:42:43,480 --> 04:42:47,760 FOR MUCIN O GLYCOPROTEINS. ONE 6467 04:42:47,760 --> 04:42:49,560 THING TAKING OFF GAINING 6468 04:42:49,560 --> 04:42:50,800 MOMENTUM AS I ENTERED THE LAB 6469 04:42:50,800 --> 04:42:56,360 WAS THE IDEA OF USING O 6470 04:42:56,360 --> 04:42:59,760 GLYCOPROTEASE. IN THE BERTUZZI 6471 04:42:59,760 --> 04:43:03,360 LAB WE SAY MUCINASES, MUCINASES 6472 04:43:03,360 --> 04:43:04,760 ARE SELECTIVE TOWARDS MUCINS IN 6473 04:43:04,760 --> 04:43:06,240 GENERAL SO THE IDEA HERE IS THAT 6474 04:43:06,240 --> 04:43:07,920 YOU CAN HAVE THIS PROTEASE, THIS 6475 04:43:07,920 --> 04:43:11,520 ONE IS FROM E. COLI SO A PROTEIN 6476 04:43:11,520 --> 04:43:12,600 PUBLISHED ON SINCE EARLY 2000S 6477 04:43:12,600 --> 04:43:14,080 AND WE STARTED EXPLORING WHAT 6478 04:43:14,080 --> 04:43:15,200 THAT MIGHT LOOK LIKE AND HOW WE 6479 04:43:15,200 --> 04:43:17,520 CAN USE IT AND THE TAKE HOME 6480 04:43:17,520 --> 04:43:18,760 MESSAGE FROM THIS PLOT IS TO SAY 6481 04:43:18,760 --> 04:43:21,000 HERE ARE RECOMBINANTLY EXPRESSED 6482 04:43:21,000 --> 04:43:22,360 MUCINS. YOU CAN SEE IF YOU 6483 04:43:22,360 --> 04:43:26,160 TREAT THEM WITH STICKY, THE NAME 6484 04:43:26,160 --> 04:43:29,520 OF THIS O GLYCOPROTEASE, YOU GET 6485 04:43:29,520 --> 04:43:30,840 CLEAVAGE YOU CAN SAY THIS 6486 04:43:30,840 --> 04:43:32,120 DEGRADING IT SOMEHOW. THE 6487 04:43:32,120 --> 04:43:34,760 PROTEASE THOUGH DOESN'T CLEAVE 6488 04:43:34,760 --> 04:43:39,280 BSA NOR DOES IT CLEAVE FET 1 A O 6489 04:43:39,280 --> 04:43:41,600 GLYCOSYLATED PROTEIN. SO A LOT 6490 04:43:41,600 --> 04:43:43,240 OF EXPLORE FOR RATION WENT INTO 6491 04:43:43,240 --> 04:43:46,000 THIS AND BEFORE I GOT INTO THE 6492 04:43:46,000 --> 04:43:49,520 LAB FIGURED OUT STCE IS A 6493 04:43:49,520 --> 04:43:51,520 MUCINASE IS SELECTIVE FOR THIS 6494 04:43:51,520 --> 04:43:52,960 MOTIF THAT EXISTS WITHIN MUCINS 6495 04:43:52,960 --> 04:43:57,680 SO THIS TXT, X CAN BE NUMBER OF OF 6496 04:43:57,680 --> 04:43:59,680 AMINO ACIDS WHERE THE 3 ANINE 6497 04:43:59,680 --> 04:44:01,520 HAS TO BE GLYCOSYLATE SOD YOU 6498 04:44:01,520 --> 04:44:02,760 CAN IMAGINE HOW DENSE THE MUCIN 6499 04:44:02,760 --> 04:44:06,440 DOMAINS ARE, THIS WOULD APPEAR A 6500 04:44:06,440 --> 04:44:08,280 LOT AND STCE CLEAVE IS HERE 6501 04:44:08,280 --> 04:44:10,360 BETWEEN SECOND AND THIRD RESIDUE 6502 04:44:10,360 --> 04:44:11,600 OF THE MOTIF IT IS NOT JUST THE 6503 04:44:11,600 --> 04:44:13,440 GLYCAN AND NOT JUST ABOUT THE 6504 04:44:13,440 --> 04:44:15,560 PEPTIDE. THESE MUCINASES REQUIRE 6505 04:44:15,560 --> 04:44:18,120 THE GLYCAN AND PEPTIDE MOTIF IN 6506 04:44:18,120 --> 04:44:20,240 COMBINATION. AND WE ARE LEARNING 6507 04:44:20,240 --> 04:44:21,640 NOW THIS EXCITEMENT IN THE FIELD 6508 04:44:21,640 --> 04:44:23,160 THERE ARE MORE MUCINASES THAN 6509 04:44:23,160 --> 04:44:26,160 JUST ONE, THERE IS THE FAMILY OF 6510 04:44:26,160 --> 04:44:28,160 O GLYCOPROTEASES THAT CAN CLEAVE 6511 04:44:28,160 --> 04:44:29,800 NON-MUCINS AS WELL. SO I WOULD 6512 04:44:29,800 --> 04:44:31,320 SAY HERE TOO TO HIGHLIGHT HOW 6513 04:44:31,320 --> 04:44:32,760 MUCH WORK IS NOW BEING DONE IN 6514 04:44:32,760 --> 04:44:36,360 THE BERTOZZI LAB, STACY AND JUDY 6515 04:44:36,360 --> 04:44:37,960 HAVE BEEN SPEARHEADING THIS 6516 04:44:37,960 --> 04:44:39,720 BEFORE AND GOT IN THE LAB AND 6517 04:44:39,720 --> 04:44:41,440 WORK TO DARK RISE NUMBER OF 6518 04:44:41,440 --> 04:44:43,240 GLYCOPROTEASES WITH ALL THESE 6519 04:44:43,240 --> 04:44:45,320 CLEAVAGE MOTIFS HERE. WE ARE 6520 04:44:45,320 --> 04:44:47,840 ALSO LOOKING AT JUDY 6521 04:44:47,840 --> 04:44:49,040 SPEARHEADING HOW TO USE INACTIVE 6522 04:44:49,040 --> 04:44:50,840 MUCINASES SO THINGS THAT HAVE 6523 04:44:50,840 --> 04:44:52,400 SPECIFICITIES FOR THESE DOMAINS 6524 04:44:52,400 --> 04:44:54,760 THAT CAN BE INACTIVATED THE F 6525 04:44:54,760 --> 04:44:55,520 THEY DON'T CLEAVE BUT STILL 6526 04:44:55,520 --> 04:44:56,760 BIND. WITH THESE YOU CAN 6527 04:44:56,760 --> 04:44:58,080 IMAGINE HOW YOU CAN USE AS 6528 04:44:58,080 --> 04:44:59,440 IMAGING REAGENT AND THINGS IN 6529 04:44:59,440 --> 04:45:01,120 THE THAT CAPACITY ALSO LOOKING 6530 04:45:01,120 --> 04:45:03,360 HOW WE CAN USE MUCINASES AS A 6531 04:45:03,360 --> 04:45:05,000 THERAPEUTIC MODALITY TOO. SO ONE 6532 04:45:05,000 --> 04:45:07,360 THING WE ARE USING INACTIVE 6533 04:45:07,360 --> 04:45:08,080 MUCINASES FOR AND I DON'T WANT 6534 04:45:08,080 --> 04:45:10,320 TO BELABOR THE POINT HOW CAN WE 6535 04:45:10,320 --> 04:45:11,840 DETERMINE WHAT MUCIN DOMAIN 6536 04:45:11,840 --> 04:45:13,200 GLYCOPROTEINS EXIST IN A SAMPLE, 6537 04:45:13,200 --> 04:45:14,960 HOW CAN WE UNDERSTAND WHAT IS 6538 04:45:14,960 --> 04:45:15,840 PRESENT IN ORDER TO CHARACTERIZE 6539 04:45:15,840 --> 04:45:18,800 WHAT IS THERE. SO WE ARE USING 6540 04:45:18,800 --> 04:45:19,880 INACTIVE MUCINASES TO THEN PULL 6541 04:45:19,880 --> 04:45:21,880 OUT MUCIN DOMAIN GLYCOPROTEINS. 6542 04:45:21,880 --> 04:45:23,440 SO THIS IS SOMETHING THAT YOU 6543 04:45:23,440 --> 04:45:26,240 CAN BASICALLY COUPLE INACTIVE 6544 04:45:26,240 --> 04:45:29,320 MUCINASE TO A BEAD AND DO SOME 6545 04:45:29,320 --> 04:45:30,480 ENRICHMENT THAT WAY AND THIS 6546 04:45:30,480 --> 04:45:32,040 SOMETHING THAT STACY SPEARHEADED 6547 04:45:32,040 --> 04:45:34,080 AND I HELPED WITH INFORMATICS ON 6548 04:45:34,080 --> 04:45:36,080 IS THE IDEA YOU CAN ENRICH MUCIN 6549 04:45:36,080 --> 04:45:38,560 DOMAIN GLYCOPROTEINS AND MUCIN 6550 04:45:38,560 --> 04:45:39,680 COMPUTATION TO UNDERSTAND THESE 6551 04:45:39,680 --> 04:45:42,000 HAVE MUCIN DOMAINS IN THESE 6552 04:45:42,000 --> 04:45:43,360 PARTICULAR SPOTS. CHALLENGE 6553 04:45:43,360 --> 04:45:45,040 THOUGH OVER THE LAST TALK ABOUT 6554 04:45:45,040 --> 04:45:46,160 BEFORE CIRCLE BACK TO THE ENDS 6555 04:45:46,160 --> 04:45:47,960 HERE IS IF YOU WANT TO 6556 04:45:47,960 --> 04:45:49,960 CHARACTERIZE THE MUCIN SITES 6557 04:45:49,960 --> 04:45:50,800 THAT O-GlcNac GLYCOSIDES 6558 04:45:50,800 --> 04:45:53,240 IN THE MUCIN DOMAINS IS 6559 04:45:53,240 --> 04:45:55,040 CHALLENGING. SO FOR SAKE OF TIME 6560 04:45:55,040 --> 04:45:56,520 I WILL SKIP THROUGH MASS SPEC 6561 04:45:56,520 --> 04:45:58,240 DETAILS OF WHAT THE 6562 04:45:58,240 --> 04:46:00,360 COMPLICATIONS ARE BUT KNOW THAT 6563 04:46:00,360 --> 04:46:03,560 HOW YOU FRAGMENT O GLYCOPEPTIDES 6564 04:46:03,560 --> 04:46:04,400 DETERMINES LEVEL OF INFORMATION 6565 04:46:04,400 --> 04:46:06,080 ACHIEVEED FROM THEM. THE CURRENT 6566 04:46:06,080 --> 04:46:07,240 TOOLS DON'T PROVIDES ACCESS TO 6567 04:46:07,240 --> 04:46:10,000 THAT INFORMATION THE WAY WE NEED 6568 04:46:10,000 --> 04:46:10,480 FOR O-GlcNac 6569 04:46:10,480 --> 04:46:11,920 GLYCOPEPTIDES. THERE'S GREAT 6570 04:46:11,920 --> 04:46:15,080 TOOLS FOR N GLYCOPEPTIDES BUT O 6571 04:46:15,080 --> 04:46:17,360 GLYCOPEPTIDES ARE MORE 6572 04:46:17,360 --> 04:46:19,000 CHALLENGING. SO WE TEAM WITH THE 6573 04:46:19,000 --> 04:46:20,280 UNIVERSITY OF WISCONSIN TO MAKE 6574 04:46:20,280 --> 04:46:22,080 A NEW SOFTWARE ACCESSIBLE AND 6575 04:46:22,080 --> 04:46:23,880 AVAILABLE TO THE COMMUNITY AND I 6576 04:46:23,880 --> 04:46:25,840 WILL MENTION HOW IT WORKS. WE 6577 04:46:25,840 --> 04:46:27,280 LOOK AT DEVELOP TYPES OF 6578 04:46:27,280 --> 04:46:28,880 FRAGMENTATION TO IDENTIFY AND 6579 04:46:28,880 --> 04:46:31,760 THEN LOCALIZE GLYCOSIDES ON 6580 04:46:31,760 --> 04:46:32,760 GLYCOPEPTIDES THAT ARE COMING 6581 04:46:32,760 --> 04:46:35,600 FROM THESE MUCIN DOMAINS. I WILL 6582 04:46:35,600 --> 04:46:36,760 MENTION FREE OPEN SOURCE 6583 04:46:36,760 --> 04:46:37,760 STICKING WITH CHEM OF UO 1 6584 04:46:37,760 --> 04:46:40,440 TRYING TO MAKE THINGS AVAILABLE 6585 04:46:40,440 --> 04:46:41,640 AND ACCESSIBLE, HAPPY THIS CAN 6586 04:46:41,640 --> 04:46:43,200 BE A TOOL PEOPLE CAN DOWNLOAD 6587 04:46:43,200 --> 04:46:44,880 AND START USING IF YOU ARE 6588 04:46:44,880 --> 04:46:46,640 INTERESTED. WE WILL GO THROUGH 6589 04:46:46,640 --> 04:46:49,000 MENTION IT IS QUICK, IDEA WE CAN 6590 04:46:49,000 --> 04:46:50,200 DO THESE SEARCHES AND FRACTIONS 6591 04:46:50,200 --> 04:46:52,760 OF A MINUTE. AND THAT WE GET AS 6592 04:46:52,760 --> 04:46:55,400 GOOD IDs AND OTHER TOOLS OUT 6593 04:46:55,400 --> 04:46:57,160 WILL SO WAY TO EXPEDITE WHERE 6594 04:46:57,160 --> 04:46:59,840 YOU CAN DO O GLYCOPROTEOMICS ON 6595 04:46:59,840 --> 04:47:01,080 MUCIN DOMAIN GLYCOPROTEINS AND 6596 04:47:01,080 --> 04:47:02,360 HELPS IN STUDIES YOU CAN SEE 6597 04:47:02,360 --> 04:47:05,960 THAT WE ARE ABLE TO NOW SHOW O 6598 04:47:05,960 --> 04:47:07,040 GLYCOPEPTIDES COMING FROM THESE 6599 04:47:07,040 --> 04:47:08,840 MUCIN ENRICHMENTS THAT WAY. THE 6600 04:47:08,840 --> 04:47:11,560 LAST THING I WILL SAY IS THAT 6601 04:47:11,560 --> 04:47:14,960 THERE ARE LOTS MORE TOOLS BEING 6602 04:47:14,960 --> 04:47:17,360 DEVELOPED IN O GLYCOPROTEOMICS 6603 04:47:17,360 --> 04:47:18,960 SPACE. THERE IS RECENT 6604 04:47:18,960 --> 04:47:20,200 EVALUATION THAT CAME OUT BUT 6605 04:47:20,200 --> 04:47:21,880 TOOLS USED DAY TO DAY WEREN'T 6606 04:47:21,880 --> 04:47:22,760 AVAILABLE WHEN THIS STUDY 6607 04:47:22,760 --> 04:47:25,040 STARTED IN 2017. SO NOW A SECOND 6608 04:47:25,040 --> 04:47:27,200 PHASE OF THIS STUDY, THAT STACY 6609 04:47:27,200 --> 04:47:30,160 FORMER POST DOC IN B HE 6610 04:47:30,160 --> 04:47:31,280 CANRTOZZI LAB NOW PROFESSOR AT 6611 04:47:31,280 --> 04:47:33,080 YALE AND CO-LEADING AS PART OF 6612 04:47:33,080 --> 04:47:35,560 THIS INITIATIVE SO WE SEW SAYS 6613 04:47:35,560 --> 04:47:37,040 ACTIVELY RECRUITING DEVELOPERS, 6614 04:47:37,040 --> 04:47:38,160 WE STOPPED THE PHASE ON SUNDAY 6615 04:47:38,160 --> 04:47:40,040 BUT IF YOU ARE INTERESTED OR 6616 04:47:40,040 --> 04:47:41,440 KNOW SOMEONE THAT IS, WE CAN 6617 04:47:41,440 --> 04:47:43,840 MAKE EXCEPTIONS. CURRENTLY WE 6618 04:47:43,840 --> 04:47:44,680 HAVE 18 TEAMS REG DEERED SO VERY 6619 04:47:44,680 --> 04:47:45,520 EXCITED ABOUT THIS AND LOOKING 6620 04:47:45,520 --> 04:47:47,760 FORWARD TO THAT. SO DON'T WANT 6621 04:47:47,760 --> 04:47:48,600 TO TAKE TOO MUCH TIME 6622 04:47:48,600 --> 04:47:50,040 ACKNOWLEDGE A LOT OF GREAT 6623 04:47:50,040 --> 04:47:51,760 PEOPLE ESPECIALLY THE UO 1 PHASE 6624 04:47:51,760 --> 04:47:54,800 THAT WAS THE TEAM BEFORE ME SO 6625 04:47:54,800 --> 04:47:55,640 KRISTINA, SHARON, CAROLYN SO 6626 04:47:55,640 --> 04:47:57,760 MANY MOTHER PEOPLE AND IF YOU 6627 04:47:57,760 --> 04:48:00,480 HAVE INTEREST IN GLYCOPROTEOMICS 6628 04:48:00,480 --> 04:48:01,960 INITIATIVE CONTACT US ASAP 6629 04:48:01,960 --> 04:48:02,760 THROUGH EMAIL OR TWITTER AS 6630 04:48:02,760 --> 04:48:04,840 WELL. THANK YOU FOR TIME. 6631 04:48:04,840 --> 04:48:05,720 [APPLAUSE] 6632 04:48:05,720 --> 04:48:10,000 NEXT SPEAKER IS DR. SHAPIRO WHO 6633 04:48:10,000 --> 04:48:14,040 WILL TALK ABOUT ST85IA6 ON SOLID 6634 04:48:14,040 --> 04:48:17,840 TUMORS. 6635 04:48:17,840 --> 04:48:18,800 I WOULD LIKE TO THANK 6636 04:48:18,800 --> 04:48:19,640 THE ORGANIZERS TO TALK TO YOU 6637 04:48:19,640 --> 04:48:23,520 ABOUT THE WORK WE HAVE DONE AND 6638 04:48:23,520 --> 04:48:25,600 FOR COMMON FUND FOR ALLOWING ME, 6639 04:48:25,600 --> 04:48:29,160 I GUESS I'M ONE OF THE 6640 04:48:29,160 --> 04:48:31,520 NON-SPECIALISTS,S NOT REALLY A 6641 04:48:31,520 --> 04:48:32,840 GLYCOBIOLOGIST BUT WE BECAME 6642 04:48:32,840 --> 04:48:34,600 INTERESTED IN IT BECAUSE WE 6643 04:48:34,600 --> 04:48:36,800 STUDY A SIALIC ACID TRANSFERASE 6644 04:48:36,800 --> 04:48:39,000 TRYING TO UNDERSTAND THE ROLE IT 6645 04:48:39,000 --> 04:48:40,240 HAS IN INHIBITING IMMUNE 6646 04:48:40,240 --> 04:48:43,760 RESPONSE THE TUMORS. SO THE ONE 6647 04:48:43,760 --> 04:48:50,240 WE STUDY IS ST85IA6 WHICH IS A 6648 04:48:50,240 --> 04:48:52,640 SILO TRANSFER' SIGH LITTLE 6649 04:48:52,640 --> 04:48:53,680 TRANSFERASE ON O LINKED 6650 04:48:53,680 --> 04:48:55,120 GLYCOPROTEINS. THE WORK WE HAVE 6651 04:48:55,120 --> 04:48:56,800 DONE IS PRODUCT OF THE ENZYME 6652 04:48:56,800 --> 04:48:58,440 BINDS TO AND INHIBITOR RECEPTOR 6653 04:48:58,440 --> 04:49:02,520 LOCATED ON IMMUNE CELLS CALLED 6654 04:49:02,520 --> 04:49:06,000 SIGLEC E, ON MACROPHAGES 6655 04:49:06,000 --> 04:49:07,560 DENDRITIC CELLS AND NEUTROPHILS 6656 04:49:07,560 --> 04:49:09,000 AND FUNCTIONS TO INHIBIT 6657 04:49:09,000 --> 04:49:11,520 CELLULAR ACTIVATION. CYCLIC E IN 6658 04:49:11,520 --> 04:49:13,840 MICE IS SIMILAR TO SIGLEC 7 AND 6659 04:49:13,840 --> 04:49:17,240 9 IN HUMANS. SIGLEC SO WE HAD 6660 04:49:17,240 --> 04:49:18,880 KNOWN THAT IF YOU ACTUALLY LOOK 6661 04:49:18,880 --> 04:49:22,920 AT SOME OF THE CANCER DATABASES, 6662 04:49:22,920 --> 04:49:23,600 THAT ST85IA6 IS OVEREXPRESSED IN 6663 04:49:23,600 --> 04:49:25,760 MANY SOME TUMORS AND IN SOME 6664 04:49:25,760 --> 04:49:27,720 TUMORS CORRELATES WITH POOR 6665 04:49:27,720 --> 04:49:29,840 RESPONSE SO WE WANTED TO 6666 04:49:29,840 --> 04:49:31,200 UNDERSTAND WHAT STATUAS DOING. 6667 04:49:31,200 --> 04:49:33,840 SO WE GENERATED PAIRED CELL 6668 04:49:33,840 --> 04:49:36,760 LINES, SO MC 38 IS A MOUSE COLON 6669 04:49:36,760 --> 04:49:39,960 CANCER CELL LINE, B 16 F 10 IS A 6670 04:49:39,960 --> 04:49:41,840 MELANOMA CELL LINE. AND WE 6671 04:49:41,840 --> 04:49:44,040 EITHER HAD ONES NEITHER CELL 6672 04:49:44,040 --> 04:49:46,920 LINES EXPRESSED ANY APPRECIABLE 6673 04:49:46,920 --> 04:49:50,320 LEVELS OF F ST85IA6 AND WE 6674 04:49:50,320 --> 04:49:52,560 GENERATED STABLE CELL LINES, 6675 04:49:52,560 --> 04:49:54,440 WHEN WE PUT ST85IA6 IN WE 6676 04:49:54,440 --> 04:49:56,600 DEMONSTRATED THEY MADE LIGANDS 6677 04:49:56,600 --> 04:49:59,600 FOR SIGLEC E SO WE USED 6678 04:49:59,600 --> 04:50:00,720 RECOMBINANT HOOKED UP TO HUMAN 6679 04:50:00,720 --> 04:50:03,400 PORTION OF IGG ONE AND COME BACK 6680 04:50:03,400 --> 04:50:05,840 WITH FLUORESCENTLY TAGGED 6681 04:50:05,840 --> 04:50:07,200 ANTI-HUMAN IGG 1 BY FLOW 6682 04:50:07,200 --> 04:50:08,400 CYTOMETRY TO BE ABLE TO QUANTIFY 6683 04:50:08,400 --> 04:50:12,400 THE BINDING. SO USING THESE CELL 6684 04:50:12,400 --> 04:50:13,960 LINES WE WERE ABLE TO 6685 04:50:13,960 --> 04:50:16,800 DEMONSTRATE BLUE IS GOING TO BE 6686 04:50:16,800 --> 04:50:18,320 WITH ST85IA6 RED IS WITHOUT. 6687 04:50:18,320 --> 04:50:21,560 THAT IN BOTH MODELS THAT THE 6688 04:50:21,560 --> 04:50:22,840 PRESENCE OF ST85IA6 ACCELERATED 6689 04:50:22,840 --> 04:50:24,560 TUMOR GROWTH AND DECREASE 6690 04:50:24,560 --> 04:50:25,720 SURVIVAL WHEN PUT INTO THE 6691 04:50:25,720 --> 04:50:32,080 FLANKS OF MICE. WE WERE ABLE TO 6692 04:50:32,080 --> 04:50:33,840 SHOW THIS IS NOT A CELL 6693 04:50:33,840 --> 04:50:35,880 INTRINSIC EFFECT ON TUMOR GROWTH 6694 04:50:35,880 --> 04:50:39,040 BUT EXTRINSIC EFFECT DUE TO 6695 04:50:39,040 --> 04:50:40,120 SIGLEC E EXPRESSION IN MOUSE AND 6696 04:50:40,120 --> 04:50:42,320 WE WERE ABLE TO DEMONSTRATE THIS 6697 04:50:42,320 --> 04:50:44,240 WORK IN VIVO AS WELL BECAUSE WE 6698 04:50:44,240 --> 04:50:46,320 GENERATED MOUSE MODELS WHERE WE 6699 04:50:46,320 --> 04:50:48,600 CAN CONTROL EXPRESSION OF 6700 04:50:48,600 --> 04:50:51,480 ST85IA6 IN A CRE DEPENDENT DOCKS 6701 04:50:51,480 --> 04:50:56,080 RELATABLE WAY SO OVEREXPRESSING 6702 04:50:56,080 --> 04:50:57,440 ST85IA6 IN EPITHELIAL CELLS IN 6703 04:50:57,440 --> 04:50:58,960 THE GUT DOESN'T EFFECT SURVIVAL 6704 04:50:58,960 --> 04:51:01,240 OF THE MOUSE. THIS IS AN 6705 04:51:01,240 --> 04:51:04,040 ESTABLISHED COLON CANCER MODEL 6706 04:51:04,040 --> 04:51:06,560 USING CRE WHICH EXPRESSES 6707 04:51:06,560 --> 04:51:08,000 EPITHELIAL CELLS OF THE SMALL 6708 04:51:08,000 --> 04:51:09,160 LARGE INTESTINE ALONG WITH 6709 04:51:09,160 --> 04:51:12,040 ACTIVATED KRAS IN A TRUNCATED 6710 04:51:12,040 --> 04:51:14,120 APC GENERATES COLON CANCER IN 6711 04:51:14,120 --> 04:51:16,120 SIX MONTHS. IF WE PUT THESE 6712 04:51:16,120 --> 04:51:18,200 TOGETHER OVEREXPRESS ST85IA6 IN 6713 04:51:18,200 --> 04:51:20,240 THIS MODEL WHAT HAPPENS IS THAT 6714 04:51:20,240 --> 04:51:21,840 TUMORIGENESIS IS ACCELERATED SO 6715 04:51:21,840 --> 04:51:22,720 YOU GET TUMOR DEVELOPMENT IN TWO 6716 04:51:22,720 --> 04:51:29,440 MONTHS. SO THE WORK WE DONE 6717 04:51:29,440 --> 04:51:33,000 INITIALLY IS AGNOSTIC, IT DOES 6718 04:51:33,000 --> 04:51:34,560 NOT IDENTIFY PROTEINS IDENTIFIED 6719 04:51:34,560 --> 04:51:35,200 BY ST85IA6 WHICH IS LEADING TO 6720 04:51:35,200 --> 04:51:37,440 THIS CHANGE IN THE IMMUNE 6721 04:51:37,440 --> 04:51:39,800 RESPONSE BUT WE HAD PAIRED CELL 6722 04:51:39,800 --> 04:51:42,040 LINES WITH AND WITHOUT ST85IA6 6723 04:51:42,040 --> 04:51:43,480 THAT ALLOW POTENTIALLY GO AFTER 6724 04:51:43,480 --> 04:51:46,160 THESE TARGETS. SO WE GOT A 6725 04:51:46,160 --> 04:51:48,480 SUPPLEMENT TO OUR NCI RO1 TO 6726 04:51:48,480 --> 04:51:49,440 IDENTIFY WHAT SOME OF THESE 6727 04:51:49,440 --> 04:51:52,840 TARGETS ARE. AND WE WORKED WITH 6728 04:51:52,840 --> 04:51:56,920 MAYANK SARASWAT WITH DR. PANDEY 6729 04:51:56,920 --> 04:52:00,240 AT MAYO FOR MASS SPEC TO LOOK AT 6730 04:52:00,240 --> 04:52:01,080 DIFFERENTIALLY SIALYLATED 6731 04:52:01,080 --> 04:52:03,880 TARGETS BETWEEN OUR ST85IA6 6732 04:52:03,880 --> 04:52:04,840 EXPRESSING AND NON-EXPRESSING 6733 04:52:04,840 --> 04:52:12,160 CELL LINES. SO THIS IS BASICALLY 6734 04:52:12,160 --> 04:52:16,360 WHAT AKILESH'S GROUP IN CELL 6735 04:52:16,360 --> 04:52:18,120 LINES WITH PULL DOWNS WITH 6736 04:52:18,120 --> 04:52:19,280 LEPTONS AND MASS SPEC. SO TODAY 6737 04:52:19,280 --> 04:52:20,840 I WOULD LIKE TO TALK BRIEFLY 6738 04:52:20,840 --> 04:52:21,920 ABOUT ONE OF THE TARGETS THAT WE 6739 04:52:21,920 --> 04:52:24,880 GOT ON THIS SCREEN. AND SO CD4 4 6740 04:52:24,880 --> 04:52:29,120 CAME UP AS A POTENTIAL TARGET 6741 04:52:29,120 --> 04:52:29,600 FOR ST85IA6 WHICH WAS 6742 04:52:29,600 --> 04:52:33,400 DIFFERENTIALLY SIALYLATED WHEN 6743 04:52:33,400 --> 04:52:37,280 ST85IA6 WAS EXPRESSED. SO IN 6744 04:52:37,280 --> 04:52:38,600 TERMS OF GLYCANS WHICH LOOK TO 6745 04:52:38,600 --> 04:52:42,640 BE MODIFIED BY ST85IA6 THEY 6746 04:52:42,640 --> 04:52:44,680 APPEAR CORE 1 AND CORE 2 O 6747 04:52:44,680 --> 04:52:47,000 GLYCANS BUT THIS DOESN'T 6748 04:52:47,000 --> 04:52:48,360 ACTUALLY TELL US WHETHER OR NOT 6749 04:52:48,360 --> 04:52:51,240 THESE GLYCANS ON CD4 4 ARE 6750 04:52:51,240 --> 04:52:54,360 GENERATING LIGANDS FOR THE 6751 04:52:54,360 --> 04:52:55,600 SIGLECS WE ARE BIOLOGICALLY 6752 04:52:55,600 --> 04:52:57,800 INTERESTED IN SO TO DO THAT I 6753 04:52:57,800 --> 04:53:07,400 DON'T KNOW HOW TO GO BACK. WE 6754 04:53:07,400 --> 04:53:08,680 USED HEAP 293 CELLS IN PART 6755 04:53:08,680 --> 04:53:11,360 BECAUSE THEY DON'T EXPRESS CD4 4 6756 04:53:11,360 --> 04:53:13,760 SO LOOK SPECIFICALLY FOR 6757 04:53:13,760 --> 04:53:17,880 CONTRIBUTION OF CD4 4 TO 6758 04:53:17,880 --> 04:53:19,320 RECOMBINANT SIGLEC BINDING, 6759 04:53:19,320 --> 04:53:21,240 THESE ARE RECOMBINANT CELLS 6760 04:53:21,240 --> 04:53:25,120 LINES SO NOT RECOMBINANT E BUT 6761 04:53:25,120 --> 04:53:27,000 SIGLEC 7 AND 9 AND DO 6762 04:53:27,000 --> 04:53:30,640 CO-TRANSFECTIONS WITH SIGH 6763 04:53:30,640 --> 04:53:31,520 CLINICAL TRANSFERASIST SO WE CAN 6764 04:53:31,520 --> 04:53:32,920 MARK THE CELLS TRANSFECTED AND 6765 04:53:32,920 --> 04:53:35,360 WITH BOTH OF THEM TOGETHER. WE 6766 04:53:35,360 --> 04:53:39,040 USE FLOW CYTOMETRY FOR THOSE -- 6767 04:53:39,040 --> 04:53:40,200 I DON'T KNOW WHY IT IS GOING 6768 04:53:40,200 --> 04:53:47,440 AUTOMATICALLY. SO FOR THOSE THAT 6769 04:53:47,440 --> 04:53:50,480 -- THIS IS GOING TO FRUSTRATE 6770 04:53:50,480 --> 04:53:52,440 ME. I DON'T KNOW WHY IT IS 6771 04:53:52,440 --> 04:53:56,840 AUTOMATICALLY ADVANCING. SO IF 6772 04:53:56,840 --> 04:54:02,120 WE LOOK AT THE FLOW SIGH TOM TRY 6773 04:54:02,120 --> 04:54:04,800 WE WILL DO IT THIS WAY. I HAVE A 6774 04:54:04,800 --> 04:54:07,760 DIFFERENT WAY. IF WE LOOK WITH 6775 04:54:07,760 --> 04:54:10,960 JUST CD4 TRANSFECTED OR CD4 4 6776 04:54:10,960 --> 04:54:12,200 PLUS ST85IA6. WHAT YOU SEE ON 6777 04:54:12,200 --> 04:54:15,680 THE X AXIS IS ACTUALLY LOOKING 6778 04:54:15,680 --> 04:54:19,040 FOR FLAG TAG CD4 4. SO CELLS 6779 04:54:19,040 --> 04:54:20,360 TRANSFECTED WILL BE BOXED ON THE 6780 04:54:20,360 --> 04:54:22,800 RIGHT, NON-TRANSFECTED CELLS 6781 04:54:22,800 --> 04:54:24,440 WITHIN THIS GROUP ARE ON THE 6782 04:54:24,440 --> 04:54:27,400 LEFT. AND WE ARE USING 6783 04:54:27,400 --> 04:54:31,680 RECOMBINANT SIC SIGLEC 7 ON Y AXIS 6784 04:54:31,680 --> 04:54:33,840 IN THE UP PER FIELD SO IF WE 6785 04:54:33,840 --> 04:54:37,440 OVEREXPRESS CD4 4 AND HEP 393 6786 04:54:37,440 --> 04:54:39,840 T-CELLS THERE IS NOT A LOT OF 6787 04:54:39,840 --> 04:54:42,000 SIGLEC BINDING. THIS IS 7 6788 04:54:42,000 --> 04:54:44,440 HOWEVER IF WE CO-EXPRESS ST85IA6 6789 04:54:44,440 --> 04:54:46,920 ALONG WITH CD4 4 YOU CAN SEE THE 6790 04:54:46,920 --> 04:54:51,360 SHIFT. SO IT INDICATES THAT CD4 6791 04:54:51,360 --> 04:54:55,040 4 BEING MODIFIED BY ST85IA6 IS 6792 04:54:55,040 --> 04:54:58,440 GENERATING A LIGAND FOR 6793 04:54:58,440 --> 04:55:00,720 RECOMBINANT SIGLEC 7. BECAUSE 6794 04:55:00,720 --> 04:55:02,920 ST85IA6 PUTS ON A SECOND SIALIC 6795 04:55:02,920 --> 04:55:05,440 ACID ON TO A PREEXISTING SIALIC 6796 04:55:05,440 --> 04:55:08,840 ACID WHAT COULD THAT FIRST 6797 04:55:08,840 --> 04:55:11,280 SIALIC ACID BE? HOW IS IT 6798 04:55:11,280 --> 04:55:13,200 ORIGINALLY PUT ON? SO WE DECIDED 6799 04:55:13,200 --> 04:55:17,840 TO LOOK AT ST 3 GAL 1 AND FT 6 6800 04:55:17,840 --> 04:55:20,680 GAL 1. SO HERE WE CO-TRANSFECT 6801 04:55:20,680 --> 04:55:23,680 CD4 4 WITH ST 3 GAL 1 WE DO SEE 6802 04:55:23,680 --> 04:55:26,040 THERE IS ENHANCEMENTS OF BINDING 6803 04:55:26,040 --> 04:55:29,400 OF RECOMBINANT SIGLEC 7. WHAT WE 6804 04:55:29,400 --> 04:55:31,520 SEE IS BOTH ARE TOGETHER. SO ST 6805 04:55:31,520 --> 04:55:35,840 3 GAL 1 ALLEN AN BOTH TOGETHER 6806 04:55:35,840 --> 04:55:40,440 YOU CAN SEE SYNERGY TO GENERATE 6807 04:55:40,440 --> 04:55:42,560 LIGANDS FOR SIGLEC 7. THIS IS 6808 04:55:42,560 --> 04:55:48,200 SPECIFIC FOR SIGLEC 7 IF WE USE 6809 04:55:48,200 --> 04:55:51,040 ST 6 GAL 1 WE DON'T SEE ANY 6810 04:55:51,040 --> 04:55:54,960 SYNERGY BETWEEN ST85IA6 ALONE 6811 04:55:54,960 --> 04:55:58,160 ABOUT ST85IA6 PLUS ST 6 GAL 1. 6812 04:55:58,160 --> 04:55:59,720 WHAT MY SLIDES WERE 6813 04:55:59,720 --> 04:56:00,880 AUTOMATICALLY FLIPPING THROUGH 6814 04:56:00,880 --> 04:56:03,520 IS THAT IT DID NOT GENERATE ANY 6815 04:56:03,520 --> 04:56:06,760 LIGANDS FOR RECOMBINANT SIGLEC 6816 04:56:06,760 --> 04:56:08,680 9. SO JUST TO SUMMARIZE IS THAT 6817 04:56:08,680 --> 04:56:14,000 WE IDENTIFIED THAT ST85IA6 6818 04:56:14,000 --> 04:56:19,040 GENERATES OR SYLATES CD4 4 OR 6819 04:56:19,040 --> 04:56:24,120 MURINE TUMOR CELLS, USING 293 6820 04:56:24,120 --> 04:56:25,600 CELLS WE CAN ST85IA6 CYLATION OF 6821 04:56:25,600 --> 04:56:28,080 CD4 4 LEADS TO GENERICS OF 6822 04:56:28,080 --> 04:56:29,920 LIGANDS FOR SIGLEC 7 BUT NOT 9 6823 04:56:29,920 --> 04:56:31,320 AND GENERATION OF LIGANDS FOR 6824 04:56:31,320 --> 04:56:34,760 SIGLEC 7 WAS ENHANCED WHEN ST 3 6825 04:56:34,760 --> 04:56:36,200 GAL 1 WAS CO-EXPRESSED WITH 6826 04:56:36,200 --> 04:56:37,960 ST85IA6 ALONG WITH CD # 4. SO 6827 04:56:37,960 --> 04:56:40,400 WITH THAT I WOULD LIKE TO THANK, 6828 04:56:40,400 --> 04:56:42,280 I DONE DO MASS SPECTROMETRY SO 6829 04:56:42,280 --> 04:56:44,840 WE REALLY DEPEND UPON 6830 04:56:44,840 --> 04:56:47,240 (INAUDIBLE) FOR DOING THE HARD 6831 04:56:47,240 --> 04:56:49,920 WORK TO IDENTIFY POTENTIAL 6832 04:56:49,920 --> 04:56:51,080 TARGETS AND THE WORKEN ST85IA6 6833 04:56:51,080 --> 04:56:53,560 IN THE LAB WAS STARTED BY DAVID 6834 04:56:53,560 --> 04:56:55,320 FRIEDMAN, RECENTLY DEFENDED AND 6835 04:56:55,320 --> 04:56:58,400 MOVED ON, HE WAS ONE LOOKING AT 6836 04:56:58,400 --> 04:57:00,920 ST85IA6 IN CANCER AND THE 6837 04:57:00,920 --> 04:57:02,920 IDENTIFICATION OF WHAT ARE THE 6838 04:57:02,920 --> 04:57:05,360 TARGETS THAT ARE BEING MODIFIED 6839 04:57:05,360 --> 04:57:08,320 BY ST85IA6 GENERATES LIGANDS FOR 6840 04:57:08,320 --> 04:57:10,040 SIGLECS HAVE BEEN TAKEN UP BY 6841 04:57:10,040 --> 04:57:11,320 ANOTHER GRADUATE STUDENT IN THE 6842 04:57:11,320 --> 04:57:12,520 LAB MEGHAN CHEN. SO WITH THAT 6843 04:57:12,520 --> 04:57:13,600 THANK YOU SO MUCH FOR YOUR 6844 04:57:13,600 --> 04:57:16,440 ATTENTION. I WOULD BE HAPPY TO 6845 04:57:16,440 --> 04:57:17,480 TALK TO ANYONE AFTERWARDS AT THE 6846 04:57:17,480 --> 04:57:18,680 BREAK ABOUT THE WORK THAT WE ARE 6847 04:57:18,680 --> 04:57:19,880 DOING. THANK YOU. 6848 04:57:19,880 --> 04:57:24,760 [APPLAUSE] 6849 04:57:24,760 --> 04:57:28,880 CYLATION 6850 04:57:28,880 --> 04:57:32,800 >> NEXT SPEAKER IS DR. DRAKE, 6851 04:57:32,800 --> 04:57:44,600 SIMPLIFIED GLYCAN WORK FLOWS. 6852 04:57:44,600 --> 04:57:46,160 >> THANKS TO THE COMMON FUND FOR 6853 04:57:46,160 --> 04:57:47,680 GIVING THIS TALK. ON BEHALF OF 6854 04:57:47,680 --> 04:57:54,400 MY COLLEAGUES, -- I SEE THE 6855 04:57:54,400 --> 04:58:01,840 SLIDE THAT -- THERE WE GO. MY 6856 04:58:01,840 --> 04:58:06,760 COLLEAGUE, IS MULTI-PI METHA AND 6857 04:58:06,760 --> 04:58:08,520 PEGGY ANGEL, WE ARE APPRECIATE 6858 04:58:08,520 --> 04:58:10,800 THE LAB ALLOWED US TO MAKE THESE 6859 04:58:10,800 --> 04:58:14,160 TOOLS. THE GRAND WAS SUCCESSFUL, 6860 04:58:14,160 --> 04:58:16,760 WE HAVE BEEN VERY HAPPY WITH IT. 6861 04:58:16,760 --> 04:58:19,640 BECAUSE WE STARTED WITH THE IDEA 6862 04:58:19,640 --> 04:58:22,840 WE WORK WITH CLINICAL SAMPLES, 6863 04:58:22,840 --> 04:58:25,640 COULD WE HAVE A METHOD, METHODS 6864 04:58:25,640 --> 04:58:26,960 WHERE WE CAN LOOK AT ANYTHING WE 6865 04:58:26,960 --> 04:58:28,960 CAN GET CLINICALLY WHETHER 6866 04:58:28,960 --> 04:58:31,840 TISSUE, BLOOD SERUM, URINE, 6867 04:58:31,840 --> 04:58:34,880 BIOFLUIDS YOU NAME IT. THINGS 6868 04:58:34,880 --> 04:58:38,240 READILY AVAILABLE BUT THEN 6869 04:58:38,240 --> 04:58:40,680 GLYCOTYPE ESSENTIALLY. SO THIS 6870 04:58:40,680 --> 04:58:43,360 WAS ADAPTED FROM OUR METHODS 6871 04:58:43,360 --> 04:58:45,600 DOING TISSUE IMAGING. AND THE 6872 04:58:45,600 --> 04:58:49,000 FACT THAT WE HAD TISSUE ON A 6873 04:58:49,000 --> 04:58:52,640 SLIDE. WE SPRAY THE TISSUE WITH 6874 04:58:52,640 --> 04:58:53,960 PENGASE TO RELEASE THE LIGANDS 6875 04:58:53,960 --> 04:58:55,760 SO WE HIT WITH MULTI-LASER AND 6876 04:58:55,760 --> 04:59:00,000 DETECT WHAT IS PRESENT IN EACH 6877 04:59:00,000 --> 04:59:01,680 SPACE. SO USUALLY WE WERE USING 6878 04:59:01,680 --> 04:59:08,120 THESE IMAGING WORK FLOWS, WE ARE 6879 04:59:08,120 --> 04:59:12,960 GOING TO SKIP THAT. TO DO THAT 6880 04:59:12,960 --> 04:59:15,480 AND THEN GENERATE THE SPECTRA 6881 04:59:15,480 --> 04:59:17,720 AND WE CAN GENERATE THE 6882 04:59:17,720 --> 04:59:19,280 ESSENTIALLY HEAT MAPS OF 6883 04:59:19,280 --> 04:59:20,160 INTENSITIES WHERE THE DIFFERENT 6884 04:59:20,160 --> 04:59:24,640 GLYCANS WORK. YOU CAN ALSO ADAPT 6885 04:59:24,640 --> 04:59:25,840 TO THINGS NOT TISSUE, OTHER 6886 04:59:25,840 --> 04:59:26,880 THINGS ON THE SLIDE. AND 6887 04:59:26,880 --> 04:59:31,760 ANTIBODY ARRAY, WHAT IF WE SPOT 6888 04:59:31,760 --> 04:59:33,960 SERUM, THINGS LIKE THAT. SO WE 6889 04:59:33,960 --> 04:59:37,040 HAVE PULLED OFF MOST OF THIS 6890 04:59:37,040 --> 04:59:40,080 NOW, STARTED WITH TISSUE, I WILL 6891 04:59:40,080 --> 04:59:41,840 SHOW EXAMPLES OF WHAT WE ARE 6892 04:59:41,840 --> 04:59:43,440 DOING WITH WORK FLOWS ON 6893 04:59:43,440 --> 04:59:45,320 CULTURED CELLS. THAT PEGGY ANGEL 6894 04:59:45,320 --> 04:59:51,240 DEVELOPED. AND MEHT SPEARHEADED 6895 04:59:51,240 --> 04:59:54,800 THE ANTIBODY ARRAY ACTIVITIES. 6896 04:59:54,800 --> 04:59:57,800 AND I HAVE SPENDING TIME ON 6897 04:59:57,800 --> 05:00:00,800 BIOFLUIDS JUST PROFILING THE 6898 05:00:00,800 --> 05:00:02,120 CLINICAL FLUIDS. THIS WAS NOT 6899 05:00:02,120 --> 05:00:04,240 ACTUALLY PART OF THE COMMON FUND 6900 05:00:04,240 --> 05:00:05,480 PROPOSAL AT THE TIME BUT 6901 05:00:05,480 --> 05:00:08,800 EVERYTHING WE DEVELOPED WITHIN 6902 05:00:08,800 --> 05:00:09,400 THE COMMON FUND SPUN NATURALLY 6903 05:00:09,400 --> 05:00:11,320 IN TO THIS SO IT WAS A NICE 6904 05:00:11,320 --> 05:00:13,440 BY-PRODUCT. WE WILL SKIP 6905 05:00:13,440 --> 05:00:16,000 THROUGH. SO WE ALWAYS HAD THE 6906 05:00:16,000 --> 05:00:20,080 PREMISE USING A MALDI INSTRUMENT 6907 05:00:20,080 --> 05:00:25,440 BASED ON HOW SIMPLE BENCH TOP 6908 05:00:25,440 --> 05:00:26,080 MALDI TRANSFORMED CLINICAL 6909 05:00:26,080 --> 05:00:29,600 MICROBIOLOGY. GLYCOTYPING OR 6910 05:00:29,600 --> 05:00:31,640 BIOTYPING OF BACTERIA IS DONE 6911 05:00:31,640 --> 05:00:34,640 NOW BY MALDI, VERY ROUTINE, 6912 05:00:34,640 --> 05:00:36,920 ESSENTIALLY A BAR CODE TYPE. 6913 05:00:36,920 --> 05:00:39,640 COULD WE DO THAT WITH GLYCANS 6914 05:00:39,640 --> 05:00:41,680 BECAUSE ESSENTIALLY THE MALDI IS 6915 05:00:41,680 --> 05:00:42,920 GIVING YOU THAT INFORMATION, 6916 05:00:42,920 --> 05:00:46,840 VERY CONSISTENT N GLYCAN PEAKS, 6917 05:00:46,840 --> 05:00:49,240 KNOWN MASSES, CAN WE DO THIS FOR 6918 05:00:49,240 --> 05:00:54,320 DIFFERENT CLINICAL CONDITIONS. 6919 05:00:54,320 --> 05:00:57,960 WE WON'T GO IN ORDER OF THE AIMS 6920 05:00:57,960 --> 05:01:01,240 BUT PEGGY PUBLISHED THE INITIAL 6921 05:01:01,240 --> 05:01:03,000 PAPER, THE SHORT VERSION YOU CAN 6922 05:01:03,000 --> 05:01:07,200 GROW ON THE SLIDE, ANY ATTACHED 6923 05:01:07,200 --> 05:01:09,040 CELL ON THE SLIDE WE CAN SHOOT 6924 05:01:09,040 --> 05:01:10,960 IT AND DO GLYCOPROFILING. IT'S A 6925 05:01:10,960 --> 05:01:16,600 METHOD PAPER AND DOESN'T TAKE 6926 05:01:16,600 --> 05:01:18,840 THAT MANY CELLS TO GET GOOD 6927 05:01:18,840 --> 05:01:22,760 SIGNAL, WE CAN GO LOW AS 10 6928 05:01:22,760 --> 05:01:27,440 DEPENDING ON CELL TYPE. WE ARE 6929 05:01:27,440 --> 05:01:28,840 GETTING TOWARD SING CELL, BASED 6930 05:01:28,840 --> 05:01:33,680 ON PEGGY'S CCRC ROOTS SHE ALSO 6931 05:01:33,680 --> 05:01:37,360 ADAPTED THE IDOL METHOD 6932 05:01:37,360 --> 05:01:39,040 INCORPORATING GLUTAMINE INTO THE 6933 05:01:39,040 --> 05:01:42,120 CELLS. I LOVE THIS BECAUSE YOU 6934 05:01:42,120 --> 05:01:45,920 CAN JUST SEE THE ONE IN YELLOW 6935 05:01:45,920 --> 05:01:48,360 REPRESENT THE HIGH MAN NOSE 6936 05:01:48,360 --> 05:01:49,360 INCORPORATED, WHILE BLUE CELLS 6937 05:01:49,360 --> 05:01:51,240 AREN'T INCORPORATING -- THAT 6938 05:01:51,240 --> 05:01:52,480 DOESN'T MEAN THEY ARE DEAD BUT 6939 05:01:52,480 --> 05:01:54,920 THEY ARE NOT CYCLING PROBABLY. 6940 05:01:54,920 --> 05:01:56,440 SO INTERESTING INFORMATION THERE 6941 05:01:56,440 --> 05:01:59,640 LOOKING AT GLYCANS. THIS ISN'T 6942 05:01:59,640 --> 05:02:03,600 A SINGLE CELL IT IS A SINGLE 6943 05:02:03,600 --> 05:02:05,400 ORGANISM BUT METHODS DEVELOPED 6944 05:02:05,400 --> 05:02:10,360 TO DO THE CELLS WE WORK WITH 6945 05:02:10,360 --> 05:02:11,880 HEATHER, STEAT AND RICH STEAT 6946 05:02:11,880 --> 05:02:15,280 ALSO IN SOUTH CAROLINA GREEN 6947 05:02:15,280 --> 05:02:17,840 WOOD KINETIC CENTER. THEY HAD 6948 05:02:17,840 --> 05:02:22,400 ZEBRAFISH MODEL OF A CDG FOR 6949 05:02:22,400 --> 05:02:25,120 PHOSPHOMANU MYTASE SO TOOK A RUN 6950 05:02:25,120 --> 05:02:28,160 AT IT, LOVE PICTURES BUT THERE 6951 05:02:28,160 --> 05:02:31,800 IS BIOLOGY HERE, IT IS WHAT YOU 6952 05:02:31,800 --> 05:02:33,720 EXPECT HIGH MANNOSE ARE 6953 05:02:33,720 --> 05:02:38,640 SUPPRESSED AND THE PHOSPHOMYTASE 6954 05:02:38,640 --> 05:02:39,760 MUTANT BUT FOR SOME OTHER REASON 6955 05:02:39,760 --> 05:02:42,280 IN THAT MUTANT THE LITTLE BY 6956 05:02:42,280 --> 05:02:46,160 SECT GLYCANS ARE VERY ELEVATED 6957 05:02:46,160 --> 05:02:47,640 SO SOME METABOLIC CONVERSATION 6958 05:02:47,640 --> 05:02:54,800 GOING ON THERE. BRIEFLY, I HAVE 6959 05:02:54,800 --> 05:02:56,600 A STUDENT WHO DECIDED TO LOOK AT 6960 05:02:56,600 --> 05:02:58,600 THREE PANCREATIC CANCER CELLS 6961 05:02:58,600 --> 05:03:00,480 LINES, ONE CONTROL, FOUR DRUG 6962 05:03:00,480 --> 05:03:04,520 TREATMENTS THREE TIME POINTS, 6963 05:03:04,520 --> 05:03:07,440 WITH HEAVY GLUTAMINE. ONCE HE 6964 05:03:07,440 --> 05:03:10,560 SORTS THIS OUT HE GRADUATES, 6965 05:03:10,560 --> 05:03:13,080 NICE PAPER TOO BUT AMAZING 6966 05:03:13,080 --> 05:03:15,760 THINGS, WE HAVE 80 GLYCANS TO 6967 05:03:15,760 --> 05:03:16,840 SORT THROUGH ACROSS CELL LINES 6968 05:03:16,840 --> 05:03:21,040 AND TIME POINTS. THERE WAS SOME 6969 05:03:21,040 --> 05:03:22,600 INTERESTING YOU LOOK 6970 05:03:22,600 --> 05:03:25,840 INTERESTINGLY THIS ONE IS 6971 05:03:25,840 --> 05:03:28,120 CONTROL. ONE EXAMPLE OF GLYCAN 6972 05:03:28,120 --> 05:03:33,800 ELEVATED IN RESPONSE TO DRUGS. 6973 05:03:33,800 --> 05:03:37,360 MOST PROFILE LIKE THIS THERE IS 6974 05:03:37,360 --> 05:03:39,480 A LAG BEFORE REINCORPORATING IT. 6975 05:03:39,480 --> 05:03:41,120 SO THAT IS ONGOING. THAT IS ONE 6976 05:03:41,120 --> 05:03:46,520 TOOL. IN THE CELLS. THE OTHER 6977 05:03:46,520 --> 05:03:49,160 ANTIBODY PANELS PUBLISHED AND 6978 05:03:49,160 --> 05:03:49,960 STANDARDIZE AND CURRENT 6979 05:03:49,960 --> 05:03:52,120 PROTOCOLS ALSO. AND THEN AS 6980 05:03:52,120 --> 05:03:53,400 PART OF THE GRANT WE ARE LOOKING 6981 05:03:53,400 --> 05:03:55,760 AT PRIMARILY THE IMMUNOGLOBULIN 6982 05:03:55,760 --> 05:03:58,200 AND THE SUBTYPES AND SAME 6983 05:03:58,200 --> 05:03:59,360 INFORMATION. THIS IS GOING TO BE 6984 05:03:59,360 --> 05:04:01,520 FOLLOWED UP BY THE NEXT SPEAKER, 6985 05:04:01,520 --> 05:04:04,840 BEFORE THE BREAK, THAT 6986 05:04:04,840 --> 05:04:07,240 COMMERCIALIZED THIS. SO THE IDEA 6987 05:04:07,240 --> 05:04:09,040 IS THAT AGAIN IF YOU PUT 6988 05:04:09,040 --> 05:04:14,600 ANTIBODY ARRAY ON A SLIDE, THEN 6989 05:04:14,600 --> 05:04:16,920 DO SAME WORK FLOW INCUBATE 6990 05:04:16,920 --> 05:04:18,240 CAPTURE THE TARGET. WE KNOW WHAT 6991 05:04:18,240 --> 05:04:21,240 IS IN EACH SPOT. IF ANTIBODY IS 6992 05:04:21,240 --> 05:04:22,640 GOOD WE KNOW WHAT TARGET IS. 6993 05:04:22,640 --> 05:04:25,760 THESE TARGETS BEING 6994 05:04:25,760 --> 05:04:28,040 GLYCOPROTEINS IN THIS CASE 6995 05:04:28,040 --> 05:04:31,040 IMMUNOGLOBULINS, IMMUNOGLOBULIN 6996 05:04:31,040 --> 05:04:33,360 Gs, AND WORK FLOW YOU SPRAY 6997 05:04:33,360 --> 05:04:39,240 THE PENGASE YOU CAN DETECT 6998 05:04:39,240 --> 05:04:42,200 GLYCANS NICE PAPER PUBLISHED, 6999 05:04:42,200 --> 05:04:45,280 LAST THREE OR FOUR MONTHS, WE 7000 05:04:45,280 --> 05:04:47,480 TURN GLYCOFIBER TYPIER. THIS IS 7001 05:04:47,480 --> 05:04:49,200 BLURRY BETWEEN ACADEMIC HATS AND 7002 05:04:49,200 --> 05:04:54,720 COMPANY HAT. THIS WE WILL 7003 05:04:54,720 --> 05:04:55,320 COMMERCIALLYZE FOR EARLY LIVER 7004 05:04:55,320 --> 05:05:02,600 DISEASE. IT IS LOOKING AT EARLY 7005 05:05:02,600 --> 05:05:03,360 STAGE FIBROSIS, EARLY STAGE 7006 05:05:03,360 --> 05:05:04,640 DISEASE FROM LATER STAGE 7007 05:05:04,640 --> 05:05:09,320 FIBROSIS. THIS IS THE THING WE 7008 05:05:09,320 --> 05:05:11,240 DO WITH EVERY ANTIBODY, THE 7009 05:05:11,240 --> 05:05:13,560 BEAUTY OF GOING WITH THE 7010 05:05:13,560 --> 05:05:15,240 ABUNDANT SERUM GLYCOPROTEIN, 7011 05:05:15,240 --> 05:05:17,440 STANDARDS ARE THERE. SO YOU CAN 7012 05:05:17,440 --> 05:05:19,880 LOOK AT STANDARD, THEN YOU CAN 7013 05:05:19,880 --> 05:05:21,880 USE NORMAL SERUM TO CHARACTERIZE 7014 05:05:21,880 --> 05:05:25,800 WHAT PROFILE ARE YOU GETTING. SO 7015 05:05:25,800 --> 05:05:27,440 STANDARD IS NOT FROM NORMAL 7016 05:05:27,440 --> 05:05:29,200 SERUM BUT IT IS CONSISTENT THIS 7017 05:05:29,200 --> 05:05:34,840 WAY. WHEN YOU LOOK AT CLINICAL 7018 05:05:34,840 --> 05:05:37,440 SCENARIO, THIS IS YOU SEE 7019 05:05:37,440 --> 05:05:40,160 SUBTYPE DIFFERENCES. GLYCANS 7020 05:05:40,160 --> 05:05:42,920 SUBTYPE YOU CAN LINK BACK TO 7021 05:05:42,920 --> 05:05:44,080 CLINICAL CONDITION. YOU CAN 7022 05:05:44,080 --> 05:05:45,800 START TO GET RAW CURVES THAT 7023 05:05:45,800 --> 05:05:48,200 HAVE VERY SIGNIFICANT CLINICAL 7024 05:05:48,200 --> 05:05:50,440 MEANING. UNFORTUNATELY THIS WAS 7025 05:05:50,440 --> 05:05:52,440 ONLY DONE ON 80 SAMPLES SO THIS 7026 05:05:52,440 --> 05:05:54,560 IS SET UP FOR MUCH LARGER 7027 05:05:54,560 --> 05:05:58,320 PROCESS. WE'LL COME BACK IN A 7028 05:05:58,320 --> 05:06:01,880 BIT. NEXT STEPS ARE TWO 7029 05:06:01,880 --> 05:06:04,880 DIRECTIONS, ONE I WILL COVER 7030 05:06:04,880 --> 05:06:09,440 SHORTLY. ABOUT HAVING SPUN THIS 7031 05:06:09,440 --> 05:06:11,360 OUT COMMERCIALLYLY. ALSO WE 7032 05:06:11,360 --> 05:06:12,960 ENVISION ON SIMPLE ANGLE TAKING 7033 05:06:12,960 --> 05:06:16,440 THE NEXT TOP TENSER RUM 7034 05:06:16,440 --> 05:06:17,880 GLYCOPROTEINS INCORPORATED INTO 7035 05:06:17,880 --> 05:06:25,040 THE WORK FLOWS. SO THAT WAS -- 7036 05:06:25,040 --> 05:06:26,800 THE LAST THING THIS ONE TOOK A 7037 05:06:26,800 --> 05:06:28,400 WHILE AND WE WENT ABOUT IT IN 7038 05:06:28,400 --> 05:06:32,840 DIFFERENT WAYS. THE IDEA IS DO 7039 05:06:32,840 --> 05:06:34,120 IMMUNE CAPTURE IN IMMUNE CELLS 7040 05:06:34,120 --> 05:06:35,600 AND GLYCOTYPE IT THE SAME WORK 7041 05:06:35,600 --> 05:06:40,440 FLOW ON THE BACK END. IT TOOK 7042 05:06:40,440 --> 05:06:44,040 SOME TIME AND DEDICATED GRADUATE 7043 05:06:44,040 --> 05:06:45,840 STUDENT JAKE DRESSMAN TO PULL IT 7044 05:06:45,840 --> 05:06:47,640 OFF BUT WHAT YOU ARE SEEING IS 7045 05:06:47,640 --> 05:06:49,880 DOES LOOK LIKE LYMPHOCYTES 7046 05:06:49,880 --> 05:06:53,880 HALFWAY TO YOU AND THEY ARE 7047 05:06:53,880 --> 05:06:54,800 ATTACHED, ON A SLIDE BECAUSE 7048 05:06:54,800 --> 05:06:56,760 THEY ARE ANTIBODY CAPTURED. 7049 05:06:56,760 --> 05:07:00,080 TURNED OUT THE KEY WAS THE 7050 05:07:00,080 --> 05:07:01,640 FIXATION DELIPID DAVIS STEP WE 7051 05:07:01,640 --> 05:07:03,720 DO A DELIPIDATION TO GET RID OF 7052 05:07:03,720 --> 05:07:07,440 THE SMALL MOLECULES AND SOME OF 7053 05:07:07,440 --> 05:07:09,600 THE SALTS THAT WASH WITH IT. 7054 05:07:09,600 --> 05:07:14,400 OPENS UP GLYCANS FOR ACCESS. IN 7055 05:07:14,400 --> 05:07:17,800 THIS CASE THE KEY WAS AFTER YOU 7056 05:07:17,800 --> 05:07:23,800 INCUBATE CELLS CAPTURED THEN DID 7057 05:07:23,800 --> 05:07:24,520 FORMALIN FIXATION TO LOCK IN. 7058 05:07:24,520 --> 05:07:30,760 ONCE WE DID THAT, IT'S GOLDEN, 7059 05:07:30,760 --> 05:07:32,640 WE HAVE DONE THINGS WITH FACT 7060 05:07:32,640 --> 05:07:35,840 SORTED CELLS YOU CAN APPLY THAT 7061 05:07:35,840 --> 05:07:38,680 AND GIVE SAME WORK FLOW SO 7062 05:07:38,680 --> 05:07:42,320 APPEARING QUITE ROBUST. WE ARE 7063 05:07:42,320 --> 05:07:44,360 STILL FRONT END BUT I WILL SHOW 7064 05:07:44,360 --> 05:07:49,240 YOU QUICK EXAMPLES. WE CAN ALSO 7065 05:07:49,240 --> 05:07:52,640 DO -- WE DID NON-LYMPHOCYTES, 7066 05:07:52,640 --> 05:07:53,680 HEPG 2 LIVER CELLS TO SHARE YOU 7067 05:07:53,680 --> 05:07:58,640 CAN DO THIS CAPTURE METHOD AND 7068 05:07:58,640 --> 05:08:03,000 TREATED WITH 2 FOLLOW -- DEOXI 7069 05:08:03,000 --> 05:08:07,320 DEOXIFOLLOW FUCOSE. AND YOU CAN 7070 05:08:07,320 --> 05:08:12,040 SEE INVERSE OF THE TREATED ONES, 7071 05:08:12,040 --> 05:08:14,640 LOSE THAT EXTRA FUCOSE AND ONES 7072 05:08:14,640 --> 05:08:15,720 THAT DON'T YOU CAN SEE BUILDING 7073 05:08:15,720 --> 05:08:20,160 UP IN THE ONE THAT WASN'T -- THE 7074 05:08:20,160 --> 05:08:22,240 INVERSE. YOU TELL I DIDN'T DO 7075 05:08:22,240 --> 05:08:27,440 THIS ONE. WE HAVE DONE GREAT 7076 05:08:27,440 --> 05:08:29,040 PROOF OF CONCEPT BEFORE GETTING 7077 05:08:29,040 --> 05:08:34,680 TO PBMCs, TAKING SPLEENS OR 7078 05:08:34,680 --> 05:08:36,240 T-CELLS LYMPHOCYTES OUT OF MOUSE 7079 05:08:36,240 --> 05:08:38,160 SPLEENS AND SORTING THEM, GOING 7080 05:08:38,160 --> 05:08:41,520 AFTER THE CD4 AND CD8 IN THAT 7081 05:08:41,520 --> 05:08:43,840 POPULATION. AGAIN YOU CAN SEE 7082 05:08:43,840 --> 05:08:45,440 WHAT I LIKE ABOUT THIS ASSAY, 7083 05:08:45,440 --> 05:08:49,600 MAKES ME SMILE IS YOU CAN SEE 7084 05:08:49,600 --> 05:08:50,840 THIS CAPTURED PUT IT UNDER SCOPE 7085 05:08:50,840 --> 05:08:55,320 AND SEE IF YOU GOT A CELL THERE. 7086 05:08:55,320 --> 05:08:57,640 SO HE CAN AGAIN LOOKING AT THE 7087 05:08:57,640 --> 05:08:59,800 CD4 AND CD8 PROOF OF CONCEPT IS 7088 05:08:59,800 --> 05:09:02,160 CLEAN. IT IS THE TYPE OF GLYCANS 7089 05:09:02,160 --> 05:09:04,480 WE SEE ACTIVELY GROWING 7090 05:09:04,480 --> 05:09:05,840 LYMPHOCYTES, THE MICE SUCKING 7091 05:09:05,840 --> 05:09:08,080 ONE Z AS WELL AS HIGH MANNOSE. 7092 05:09:08,080 --> 05:09:10,200 THE CBs ARE IN PLAY BUT THEY 7093 05:09:10,200 --> 05:09:12,160 ARE GETTING GOOD. WE ARE 7094 05:09:12,160 --> 05:09:13,440 GETTING INTO THE LESS THAN 10 7095 05:09:13,440 --> 05:09:14,840 FOR THE MOST PART SO WE LIKE 7096 05:09:14,840 --> 05:09:16,720 WHERE THIS ASSAY IS GOING. FOR 7097 05:09:16,720 --> 05:09:18,960 THE MOST PART. THEN GIVING 7098 05:09:18,960 --> 05:09:23,280 CREDIT HE ADAPTED A CELL 7099 05:09:23,280 --> 05:09:26,320 COUNTING PROGRAM ADAPTED TO 7100 05:09:26,320 --> 05:09:30,840 THIS. SO AGAIN THE ASSAYS WE ARE 7101 05:09:30,840 --> 05:09:35,120 GOING TO HAVE INTENSITY DATA 7102 05:09:35,120 --> 05:09:36,840 MORALIZE CELL COUNT SO EXTRA 7103 05:09:36,840 --> 05:09:39,640 LAYER OF QUANTITATION, IS GOING 7104 05:09:39,640 --> 05:09:44,640 B TO INTERESTING DOWN THE LINE. 7105 05:09:44,640 --> 05:09:45,800 SO THERE IS A LOT YOU CAN DO 7106 05:09:45,800 --> 05:09:46,840 WITH THIS THINGS, LOT OF THE 7107 05:09:46,840 --> 05:09:48,720 TARGETS SO WE WILL GO TO BLOOD 7108 05:09:48,720 --> 05:09:51,840 AND PBMCs FROM CLINICAL 7109 05:09:51,840 --> 05:09:52,960 SAMPLES, THEY ARE WORKING ON 7110 05:09:52,960 --> 05:09:57,200 ORGAN ASSOCIATIONS WITH THE 7111 05:09:57,200 --> 05:09:58,920 MOUSE, REALITY IS, PRETTY MUCH 7112 05:09:58,920 --> 05:10:01,040 3-D ASSAYS WE DISCUSSED FOR THE 7113 05:10:01,040 --> 05:10:02,720 MOST PART, IT IS ALL GOING TO GO 7114 05:10:02,720 --> 05:10:06,640 TO IMMUNOTHERAPY. DOWN THE 7115 05:10:06,640 --> 05:10:08,040 LINE, THAT IS WHERE WE APPLY 7116 05:10:08,040 --> 05:10:12,000 THESE. I WANTED TO THANK COMMON 7117 05:10:12,000 --> 05:10:16,280 FUND, WE HAVE ALL THESE WORK 7118 05:10:16,280 --> 05:10:17,960 FLOWS, WE HAVE TURNED INTO A 7119 05:10:17,960 --> 05:10:21,800 PLATFORM THIS IS ESSENTIALLY A 7120 05:10:21,800 --> 05:10:23,520 PLATFORM TECHNOLOGY NOW WHATEVER 7121 05:10:23,520 --> 05:10:26,480 WE CAN STICK ON WHATEVER WE CAN 7122 05:10:26,480 --> 05:10:28,360 STICK ON GLASS THIS BACK END IS 7123 05:10:28,360 --> 05:10:30,560 A PLATFORM WE KNOW WE CAN 7124 05:10:30,560 --> 05:10:36,640 GENERATE THE GLYCANS OFF OF IT. 7125 05:10:36,640 --> 05:10:39,440 THE GOAL BEING WE WANT TO DO 7126 05:10:39,440 --> 05:10:41,760 PATIENT SPECIFIC GLYCOTYPE OR 7127 05:10:41,760 --> 05:10:43,600 TEST FOR STRESSING CANCER WORLD 7128 05:10:43,600 --> 05:10:45,240 YOU CAN SEE WE GET ALL THESE 7129 05:10:45,240 --> 05:10:49,080 TYPES OF SAMPLES. AND GLYCOTYPE 7130 05:10:49,080 --> 05:10:54,480 IT. BRIEF OUTREACH COMPONENTS, 7131 05:10:54,480 --> 05:10:57,280 DURING THE PANDEMIC WE STARTED 7132 05:10:57,280 --> 05:10:58,960 COLLABORATION THAT FORM 7133 05:10:58,960 --> 05:11:02,000 GLYCOGLYCOMICS CENTER OF 7134 05:11:02,000 --> 05:11:03,840 EXCELLENCE WITH THE 7135 05:11:03,840 --> 05:11:04,640 MANUFACTURERS OF THE MASS SPEC 7136 05:11:04,640 --> 05:11:06,080 WE USE. WE ARE DEDICATING THIS 7137 05:11:06,080 --> 05:11:08,600 FOR TRAINING, TO TRY THE GET 7138 05:11:08,600 --> 05:11:12,480 ASSAYS OUT INTO OTHER PEOPLE'S 7139 05:11:12,480 --> 05:11:15,640 HANDS. AND IN THAT REGARD, THE 7140 05:11:15,640 --> 05:11:17,640 NUMBER AFTER HURRICANE SEASON IS 7141 05:11:17,640 --> 05:11:19,600 OVER ANYBODY WANTS SOME HANDS ON 7142 05:11:19,600 --> 05:11:20,960 RAINING WITH ANY ONE OF THESE 7143 05:11:20,960 --> 05:11:22,160 TYPES OF APPROACHES, WE ARE 7144 05:11:22,160 --> 05:11:26,520 GOING TO DO A SMALL GROUP 7145 05:11:26,520 --> 05:11:29,760 IMAGING WORKSHOP HANDS ON 7146 05:11:29,760 --> 05:11:31,840 METHODOLOGY THAT CHARLESTON 7147 05:11:31,840 --> 05:11:34,840 CHARLES DID IN OUR PLACE. WHERE 7148 05:11:34,840 --> 05:11:37,320 WE ARE GOING CONTINUED OUTREACH, 7149 05:11:37,320 --> 05:11:40,520 AGAIN, THE IDEA OF GETTING TOOLS 7150 05:11:40,520 --> 05:11:43,480 OUT THERE, THAT COLLABORATIVE 7151 05:11:43,480 --> 05:11:46,240 CENTER HELPS ON THE TRAINING 7152 05:11:46,240 --> 05:11:51,200 PART, WE HAVE BEEN WORKING WITH 7153 05:11:51,200 --> 05:11:58,880 GLYGAN ON PROJECTS, KIOKO ON 7154 05:11:58,880 --> 05:12:02,440 DIFFERENT GLYCO 2 CAN, WORK FLOW 7155 05:12:02,440 --> 05:12:05,160 PUSH BUTTON FOR ACCESS TO 7156 05:12:05,160 --> 05:12:06,480 GLYCANS AN COMPANY IS ALSO 7157 05:12:06,480 --> 05:12:09,040 WORKING ON OUTREACH PART. A LOT 7158 05:12:09,040 --> 05:12:11,200 OF TOOLS WE ARE GOING TOWARD NIH 7159 05:12:11,200 --> 05:12:13,920 TISSUE MAPPING INITIATIVES, 7160 05:12:13,920 --> 05:12:17,320 TRYING TO INTEGRATE ALL THIS 7161 05:12:17,320 --> 05:12:20,920 GLYCOMIC IN SINGLE CELL OMICS IN 7162 05:12:20,920 --> 05:12:24,360 THESE CLINICAL SAMPLE TYPE 7163 05:12:24,360 --> 05:12:27,240 PROJECTS. AND REALLY EXPAND ON 7164 05:12:27,240 --> 05:12:29,440 THE CLINICAL BIOFLUID ANGLE. WE 7165 05:12:29,440 --> 05:12:31,120 ARE WORKING WITH PARTNERS AT FDA 7166 05:12:31,120 --> 05:12:36,280 ON COVID SAMPLES, AND THEN GOT 7167 05:12:36,280 --> 05:12:38,800 THIS PLATFORM CHANGE ENZYME, 7168 05:12:38,800 --> 05:12:42,120 CHANGE SIGNAL, IF WE DON'T USE 7169 05:12:42,120 --> 05:12:43,160 PENGASE WE HAVE THIS PLATFORM IN 7170 05:12:43,160 --> 05:12:47,240 PLACE TO LOOK FOR OTHER TARGETS. 7171 05:12:47,240 --> 05:12:50,720 WE HAVE FASCINATING DATA THAT 7172 05:12:50,720 --> 05:12:52,280 REMOAN SIMON DEVELOPED USING 7173 05:12:52,280 --> 05:12:54,280 AMYLASES AND GLYCOGEN, 7174 05:12:54,280 --> 05:12:58,400 FASCINATING STUFF AND TISSUES. 7175 05:12:58,400 --> 05:13:01,360 STACY MALAKE MENTIONED THE 7176 05:13:01,360 --> 05:13:04,840 GLYCOPROTEASES MUCINASES ARE 7177 05:13:04,840 --> 05:13:06,320 STABLE AND JUST PERFORM 7178 05:13:06,320 --> 05:13:07,400 EXCEPTIONALLY WELL ON TISSUE. 7179 05:13:07,400 --> 05:13:08,720 GOT REALLY NICE DATA MANY THAT 7180 05:13:08,720 --> 05:13:12,240 REGARD.REGARD. , THEN TALKING WITH 7181 05:13:12,240 --> 05:13:16,560 CHRIS NEW ENGLAND BIOLABS, HE'S 7182 05:13:16,560 --> 05:13:18,840 GOT A LOT OF BELIE COST DAYSES, 7183 05:13:18,840 --> 05:13:21,000 SOME COMMERCIALLY AVAILABLE AND 7184 05:13:21,000 --> 05:13:22,320 SOME AREN'T WHEN STRIKING 7185 05:13:22,320 --> 05:13:23,320 PARTNERSHIPS TO START 7186 05:13:23,320 --> 05:13:25,840 INTEGRATING THIS IN TO THESE 7187 05:13:25,840 --> 05:13:28,120 DIFFERENT WORK FLOWS. AND SEE 7188 05:13:28,120 --> 05:13:32,240 WHERE IT GOES. A LOT OF 7189 05:13:32,240 --> 05:13:33,600 ACKNOWLEDGMENTS. WE HAVE 7190 05:13:33,600 --> 05:13:34,240 PRIMARILY STUDENTS WHO HAD DONE 7191 05:13:34,240 --> 05:13:38,400 A LOT OF GREAT WORK. AND MY 7192 05:13:38,400 --> 05:13:40,120 COLLEAGUES PEGGY AND THE COMMON 7193 05:13:40,120 --> 05:13:42,840 FUND AND ANAND, AND COMMON FUND 7194 05:13:42,840 --> 05:13:47,000 FUNDING. NOW, FOR THE NEXT 7195 05:13:47,000 --> 05:13:50,760 SPEAKER -- BEFORE THE NEXT 7196 05:13:50,760 --> 05:13:53,840 SPEAKER. I'M HAPPY TO INTRODUCE 7197 05:13:53,840 --> 05:13:58,240 THE NEXT SPEAKER, SEEMS LIKE I 7198 05:13:58,240 --> 05:14:03,800 HAVE KNOWN HIM A LONG TIME. YEAH 7199 05:14:03,800 --> 05:14:11,200 OKAY IT IS ME. SEE, I CHANGED. 7200 05:14:11,200 --> 05:14:12,360 YOU WON'T GET THE PITCH, MAYBE A 7201 05:14:12,360 --> 05:14:15,240 LITTLE BUT THIS IS HOW WE ARE 7202 05:14:15,240 --> 05:14:18,960 TRYING TO COMMERCIALIZE AIM ONE, 7203 05:14:18,960 --> 05:14:23,680 THE GLYCAN ANTIBODY ARRAYS. 7204 05:14:23,680 --> 05:14:25,160 ANAND ALWAYS TO HIS CREDIT HAS 7205 05:14:25,160 --> 05:14:27,680 BEEN A LIVER DISEASE PERSON. I 7206 05:14:27,680 --> 05:14:30,880 DON'T HAVE TO FOCUS, I CAN DO A 7207 05:14:30,880 --> 05:14:33,440 LOT OF DIFFERENT ONES. I ALWAYS 7208 05:14:33,440 --> 05:14:38,080 LOOK FOR THE NEXT TISSUE. THAT 7209 05:14:38,080 --> 05:14:40,960 IS HOW WE ARE GOING TO TARGET 7210 05:14:40,960 --> 05:14:42,560 THAT AGAIN WITH SERUM IS IDEAL 7211 05:14:42,560 --> 05:14:44,120 FOR LOOKING AT LIVER DISEASE, IT 7212 05:14:44,120 --> 05:14:47,240 IS ALWAYS THERE. AND IRONICALLY 7213 05:14:47,240 --> 05:14:51,680 WE ARE LOOKING AT 7214 05:14:51,680 --> 05:14:53,760 IMMUNOGLOBULINS BUT THE 7215 05:14:53,760 --> 05:14:54,320 HEPATOCELLULAR CARCINOMA IN 7216 05:14:54,320 --> 05:14:56,840 LIVER DISEASE IN GENERAL IS 7217 05:14:56,840 --> 05:14:57,640 BUCKLING ALL THE TRENDS AS FAR 7218 05:14:57,640 --> 05:15:02,680 AS DECREASING INCIDENCE. IT IS 7219 05:15:02,680 --> 05:15:04,960 GETTING WORSE AND ANAND WAS AT A 7220 05:15:04,960 --> 05:15:06,400 MEETING THIS WEEK AND IT IS 7221 05:15:06,400 --> 05:15:09,160 ACCELERATING SO THERE'S ALL 7222 05:15:09,160 --> 05:15:09,880 RIGHT REJECTIONS ABOUT LIVER 7223 05:15:09,880 --> 05:15:10,880 CANCER INCIDENCE BUT IT IS 7224 05:15:10,880 --> 05:15:13,880 GETTING WORSE QUICKER. AND WE 7225 05:15:13,880 --> 05:15:16,120 ARE ALL GLYCOBIOOLOGISTS HERE, 7226 05:15:16,120 --> 05:15:17,600 USUALLY WE THINK LIVER CANCER 7227 05:15:17,600 --> 05:15:22,120 AND HEPATITIS B OR C IS ALWAYS 7228 05:15:22,120 --> 05:15:24,800 PART OF THE GLYCO COMMUNITY, IT 7229 05:15:24,800 --> 05:15:26,640 IS FATTY LIVER NOW. IT IS STILL 7230 05:15:26,640 --> 05:15:28,320 ENDS UP GETTING TO THE SAME 7231 05:15:28,320 --> 05:15:30,240 PATHWAY AS FIBROSIS CIRRHOSIS 7232 05:15:30,240 --> 05:15:32,720 LIVER CANCER BUT MOST OF THE 7233 05:15:32,720 --> 05:15:34,040 INCIDENCE ESPECIALLY IN U.S. AND 7234 05:15:34,040 --> 05:15:36,280 LOT OF EUROPE IS NOW FATTY LIVER 7235 05:15:36,280 --> 05:15:39,200 AND THAT IS WHERE WE ARE 7236 05:15:39,200 --> 05:15:41,840 TARGETING ASSAYS. THE TEAM IS 7237 05:15:41,840 --> 05:15:44,280 MYSELF ANAND AND PEGGY, FORMER 7238 05:15:44,280 --> 05:15:45,840 GRADUATE STUDENT DANIELLE WE 7239 05:15:45,840 --> 05:15:49,040 WERE FOUNDERS. ANYONE WE KNEW A 7240 05:15:49,040 --> 05:15:51,240 COLLEAGUE FROM THE IMAGING 7241 05:15:51,240 --> 05:15:53,840 COMMUNITY STEVEN WHO IS 25 YEARS 7242 05:15:53,840 --> 05:15:56,720 AT GLAXOSMITHKLINE IN PKPD SO HE 7243 05:15:56,720 --> 05:15:58,680 IS A GOOD ASSAY DEVELOPER THAT 7244 05:15:58,680 --> 05:16:02,640 WE HAVE GONE ON FOR -- HE 7245 05:16:02,640 --> 05:16:07,680 RETIRED THEN THE NEXT DAY WE GOT 7246 05:16:07,680 --> 05:16:09,040 INVOLVED. IS HOW THAT WORKED OUT 7247 05:16:09,040 --> 05:16:11,000 SO WE ARE USING ANN BODY ARRAY, 7248 05:16:11,000 --> 05:16:14,040 THERE IS A LOT OF GOOD ASSAY, 7249 05:16:14,040 --> 05:16:15,440 BUILDING ON THE LONG HISTORY O 7250 05:16:15,440 --> 05:16:18,600 OF LOOKING AT IMMUNOGLOBULIN G 7251 05:16:18,600 --> 05:16:20,040 GLYCOSYLATION, WE DIDN'T HAVE TO 7252 05:16:20,040 --> 05:16:22,600 INVENT THAT PART THERE'S A LOT 7253 05:16:22,600 --> 05:16:23,600 OF STUDIES OUTS THERE AND 7254 05:16:23,600 --> 05:16:26,000 SAMPLES THAT HAVE GONE THROUGH, 7255 05:16:26,000 --> 05:16:28,240 MAINLY IN EUROPE BUT ANAND WAS 7256 05:16:28,240 --> 05:16:29,640 PART OF EARLY STUDIES IN THAT 7257 05:16:29,640 --> 05:16:33,920 REGARD TOO. SO REALLY WHAT WE 7258 05:16:33,920 --> 05:16:35,960 HAVE SEEN IS WE WANT TO MAKE IT 7259 05:16:35,960 --> 05:16:41,720 -- THERE IS THE PITCH PART, EASY 7260 05:16:41,720 --> 05:16:42,880 FASTER COST EFFECTIVE AND 7261 05:16:42,880 --> 05:16:45,440 BETTER, THAT'S CURRENT COST 7262 05:16:45,440 --> 05:16:46,680 BETTER THAN CURRENT ONES BUT IT 7263 05:16:46,680 --> 05:16:49,480 COMES DOWN THE BAR CODE CONCEPT, 7264 05:16:49,480 --> 05:16:52,280 THE -- I LIKE TO THINK OF IT THE 7265 05:16:52,280 --> 05:16:55,400 IMMUNOGLOBULIN G GLYCANS AS 7266 05:16:55,400 --> 05:16:57,280 GOLDILOX TYPE ONES. THESE ARE 7267 05:16:57,280 --> 05:16:58,760 JUST RIGHT ONES, RIGHT? THE ONES 7268 05:16:58,760 --> 05:16:59,920 THAT THERAPEUTIC COMPANIES HAVE 7269 05:16:59,920 --> 05:17:02,920 TO HAVE ON THEIR ANTIBODY SO 7270 05:17:02,920 --> 05:17:04,160 THEY DON'T WORRY ABOUT IT AND 7271 05:17:04,160 --> 05:17:07,160 THEN THERE IS THE OTHER ONES. 7272 05:17:07,160 --> 05:17:10,960 THAT CAUSE PROBLEMS. THAT IS 7273 05:17:10,960 --> 05:17:14,360 REALLY MEASURING, WE DID HAVE 7274 05:17:14,360 --> 05:17:17,800 THIS PUBLISHED THIS IS LEADING 7275 05:17:17,800 --> 05:17:19,240 TO FUNDING WE HAVE GOT ANOTHER 7276 05:17:19,240 --> 05:17:22,840 PHASE 1 THAT STARTED MONTH AGO. 7277 05:17:22,840 --> 05:17:28,560 IF YOU CAN DO EARLY TO EARLY 7278 05:17:28,560 --> 05:17:31,920 LIVER DISEASE LATE STAGE 7279 05:17:31,920 --> 05:17:34,040 FIBROSIS, YOU CAN ALSO DO 7280 05:17:34,040 --> 05:17:36,000 CONTINUED ACCESS TO LIVER CANCER 7281 05:17:36,000 --> 05:17:38,400 HCC. SO WORKING ACROSS THAT 7282 05:17:38,400 --> 05:17:40,240 WHOLE SPECTRUM. AND WE HAVE 7283 05:17:40,240 --> 05:17:42,800 REALLY GOOD SEED FUNDING FROM 7284 05:17:42,800 --> 05:17:43,920 BRUKER, OWNERSHIP IN THE 7285 05:17:43,920 --> 05:17:45,960 COMPANY, SO WE WENT FROM LLC 7286 05:17:45,960 --> 05:17:47,280 WHICH LAST TIME WE HAD A MEETING 7287 05:17:47,280 --> 05:17:50,520 WE WERE LLC TO INC BECAUSE WE 7288 05:17:50,520 --> 05:17:55,000 HAD TO. SO INTERESTING PROCESS 7289 05:17:55,000 --> 05:17:58,800 ON ITSELF. GETTING BACK TO THE 7290 05:17:58,800 --> 05:18:03,120 TOOL PART. PART. WHAT WE ARE DOING 7291 05:18:03,120 --> 05:18:05,480 WITH SBIR IS REALLY OPTIMIZING 7292 05:18:05,480 --> 05:18:08,640 THE SLIDE CHEMISTRY SO THE SLIDE 7293 05:18:08,640 --> 05:18:10,080 CHEMISTRY P CAPTURE ANTIBODY, 7294 05:18:10,080 --> 05:18:13,640 AMOUNT OF ANTIBODY, QCING IT TO 7295 05:18:13,640 --> 05:18:16,240 THE END, APPARENTLY YOU DO 7296 05:18:16,240 --> 05:18:17,280 CLINICAL ASSAY SUPPOSED TO LOOK 7297 05:18:17,280 --> 05:18:18,640 THE SAME EVERY TIME YOU USE IT. 7298 05:18:18,640 --> 05:18:23,960 WHO KNEW. KIDDING.KIDDING. KIDDING. KIDDIN G. ANYWAY WE 7299 05:18:23,960 --> 05:18:26,520 ARE WORKING ON THE QC PART BUT 7300 05:18:26,520 --> 05:18:27,840 THAT SPINS OFF TO OTHER 7301 05:18:27,840 --> 05:18:30,160 TECHNOLOGY BECAUSE WE GO WITH, G 7302 05:18:30,160 --> 05:18:31,920 PANELS BUT CAN INCORPORATE OTHER 7303 05:18:31,920 --> 05:18:34,240 ANTIBODY TARGETS. THIS 7304 05:18:34,240 --> 05:18:35,080 POTENTIALLY COULD BE SOMETHING 7305 05:18:35,080 --> 05:18:38,720 WE SELL FRANKLY AS FAR AS ARRAY 7306 05:18:38,720 --> 05:18:41,440 FOR THIS. AND WE ARE WORKING ON 7307 05:18:41,440 --> 05:18:44,520 DATA ANALYSIS AND ALGORITHM SIDE 7308 05:18:44,520 --> 05:18:45,480 AND OUTSOURCING THESE WITH 7309 05:18:45,480 --> 05:18:46,640 DIFFERENT COMPANIES TO TO GET 7310 05:18:46,640 --> 05:18:51,640 THAT SO CLINICAL READ OUT PART. 7311 05:18:51,640 --> 05:18:55,560 BUSINESS MODEL WISE IS LINKED TO 7312 05:18:55,560 --> 05:18:58,040 THE PROGRAM. WE ARE NOT 7313 05:18:58,040 --> 05:18:59,200 TARGETING GLYCOBIOLOGIST FOR THE 7314 05:18:59,200 --> 05:19:01,240 ASSAY. THESE ARE CLINICIANS AND 7315 05:19:01,240 --> 05:19:03,800 CLIA LABS. FARTHEST AWAY YOU 7316 05:19:03,800 --> 05:19:06,120 CAN GET FROM GLYCOBIOLOGISTS, SO 7317 05:19:06,120 --> 05:19:12,200 WE WILL SEE HOW THAT GOES. THIS 7318 05:19:12,200 --> 05:19:15,880 IS THE LITTLE BENCH TOP THAT IS 7319 05:19:15,880 --> 05:19:20,840 SMALL, MUCH SMALLER THAN THIS 7320 05:19:20,840 --> 05:19:22,440 PROTEOME. WE CAN DO SERUM 7321 05:19:22,440 --> 05:19:25,800 PROFILING. JUST TOTAL GLYCAN 7322 05:19:25,800 --> 05:19:29,080 PROFILE SO THE LITTLE BENCH TOP 7323 05:19:29,080 --> 05:19:34,200 ONE COMPARED TO THOUSAND TIMES 7324 05:19:34,200 --> 05:19:37,080 BETTERS ARE SOLUTION SMART FLEX. 7325 05:19:37,080 --> 05:19:39,120 IT IS MORE SATISFYING, DOESN'T 7326 05:19:39,120 --> 05:19:41,240 MATTER WHICH IS IT, WE ARE HAPPY 7327 05:19:41,240 --> 05:19:44,040 WITH THIS. IN PRINCIPLE THIS 7328 05:19:44,040 --> 05:19:45,840 BENCH TOP CAN BE USED FOR BAR 7329 05:19:45,840 --> 05:19:56,120 CARD PROFILING. WE DO MARK THE 7330 05:19:56,120 --> 05:19:57,440 KIT TO DO IMAGING BUT THAT WAS 7331 05:19:57,440 --> 05:19:58,680 MAINLY BECAUSE WE ARE PART OF 7332 05:19:58,680 --> 05:20:00,280 THE KIT ANYWAY WHEN WE DID THE 7333 05:20:00,280 --> 05:20:01,680 CLINICAL ASSAY. SO WE HAVE SOLD 7334 05:20:01,680 --> 05:20:05,040 A FEW OF THOSE. IT IS NOT THE 7335 05:20:05,040 --> 05:20:08,800 BUSINESS MODEL. AND WE DO HAVE 7336 05:20:08,800 --> 05:20:09,960 CUSTOMIZE ARRAYS COMING DOWN THE 7337 05:20:09,960 --> 05:20:14,360 PIKE WHICH BOTH OF THESE MAY HIT 7338 05:20:14,360 --> 05:20:16,160 THE MORE COMMERCIAL SPACE AND 7339 05:20:16,160 --> 05:20:17,640 THAT IS CLINICAL ALSO IN SPIRIT 7340 05:20:17,640 --> 05:20:21,240 OF COMMON FUND. I WILL STOP 7341 05:20:21,240 --> 05:20:21,840 WILL. 7342 05:20:21,840 --> 05:20:25,720 [APPLAUSE] 7343 05:20:25,720 --> 05:20:26,960 I WILL STOP THERE. 7344 05:20:26,960 --> 05:20:28,240 >> GREAT. 7345 05:20:28,240 --> 05:20:32,640 [APPLAUSE] 7346 05:20:32,640 --> 05:20:35,320 >> SO WE HAVE A SHORT BREAK. WE 7347 05:20:35,320 --> 05:20:42,160 COME BACK HERE AT 3:50. AT 3:50 7348 05:20:42,160 --> 05:20:44,840 DR. EDWARDS IS HERE. HE WILL 7349 05:20:44,840 --> 05:20:50,440 GIVE HIS TALK. 7350 05:20:50,440 --> 05:20:52,160 >> OUR NEXT SPEAKER IS DR. 7351 05:20:52,160 --> 05:20:54,240 NATHAN EDWARDS AND HE'S GOING TO 7352 05:20:54,240 --> 05:20:57,840 TALK ABOUT THE WORK THAT HE AND 7353 05:20:57,840 --> 05:20:58,480 DR. GOLDMAN HAVE BEEN WORKING 7354 05:20:58,480 --> 05:21:03,720 ON. 7355 05:21:03,720 --> 05:21:05,920 >> I WANT TO TALK TO YOU ABOUT 7356 05:21:05,920 --> 05:21:07,400 THE WORK THAT ROD AND I WORKED 7357 05:21:07,400 --> 05:21:13,280 ON TO TRY TO PROVE OUT LCMS 7358 05:21:13,280 --> 05:21:17,080 ANALYSIS STRATEGY FOR SITE 7359 05:21:17,080 --> 05:21:21,480 SPECIFIC PROTEIN GLYCOFORMS. THE 7360 05:21:21,480 --> 05:21:23,520 PROMISE HERE THAT WE SAW TO 7361 05:21:23,520 --> 05:21:27,080 REALIZE WAS THIS IDEA THAT WE 7362 05:21:27,080 --> 05:21:30,120 COULD RUN A DIA STAR WORK FLOW 7363 05:21:30,120 --> 05:21:32,520 FOR QUANTIFICATION OF 7364 05:21:32,520 --> 05:21:35,040 GLYCOPEPTIDES USING THE SAME 7365 05:21:35,040 --> 05:21:35,960 ANALYSIS STRATEGY BEING 7366 05:21:35,960 --> 05:21:40,600 SUCCESSFUL FOR PROTEOMICS. 7367 05:21:40,600 --> 05:21:45,160 ENABLING US TO GET SENSITIVE 7368 05:21:45,160 --> 05:21:48,680 COMPREHENSIVE QUANTITATION OF 7369 05:21:48,680 --> 05:21:50,000 GLYCOPEPTIDES AND DR. BY 7370 05:21:50,000 --> 05:21:51,520 UNDERSTANDING NOT ONLY WHAT 7371 05:21:51,520 --> 05:21:53,040 GLYCANS ARE OBSERVED BUT WHERE 7372 05:21:53,040 --> 05:21:58,720 ON PARTICULAR PROTEIN. RON'S 7373 05:21:58,720 --> 05:22:00,240 LAB IS RESPONSIBLE FOR PROTOCOLS 7374 05:22:00,240 --> 05:22:01,840 AND ANALYTICAL WORK FLOWS OF THE 7375 05:22:01,840 --> 05:22:04,040 PART OF THIS ANALYSIS. MY LAB 7376 05:22:04,040 --> 05:22:05,600 IS RESPONSIBLE FOR INFORMATICS 7377 05:22:05,600 --> 05:22:07,840 SIDE OF THINGS. TRYING TO 7378 05:22:07,840 --> 05:22:09,440 CONSTRUCT GLYCOPEPTIDE SPECTRA 7379 05:22:09,440 --> 05:22:10,720 AND TRANSITION LIBRARIES WHICH 7380 05:22:10,720 --> 05:22:11,960 ARE NECESSARY FOR THE DIA 7381 05:22:11,960 --> 05:22:14,760 ANALYSIS. AND AS A SIDE BENEFIT 7382 05:22:14,760 --> 05:22:18,400 OF THAT, WHAT WE HAVE IS QUITE A 7383 05:22:18,400 --> 05:22:20,480 SIGNIFICANT LIBRARY OF 7384 05:22:20,480 --> 05:22:23,080 GLYCOPEPTIDES WITH THIS SPECTRAL 7385 05:22:23,080 --> 05:22:26,040 EVIDENCE. THAT PROVIDE SPECIFIC 7386 05:22:26,040 --> 05:22:27,960 DIRECT EVIDENCE OF EXISTENCE OF 7387 05:22:27,960 --> 05:22:30,120 THESE GLYCANS AN SPECIFIC SPOT 7388 05:22:30,120 --> 05:22:32,960 ON A PROTEIN, WITH ENOUGH 7389 05:22:32,960 --> 05:22:33,960 EVIDENCE YOU CAN DECIDE WHETHER 7390 05:22:33,960 --> 05:22:38,960 YOU BELIEVE THEM YOURSELF. THIS 7391 05:22:38,960 --> 05:22:40,280 IS WEBSITE THAT IS CONSTRUCTED 7392 05:22:40,280 --> 05:22:41,120 TO DAYS PLAY THIS 7393 05:22:41,120 --> 05:22:44,920 INFORMATIONMENT -- DISPLAY IN 7394 05:22:44,920 --> 05:22:47,240 INFORMATION. GPT WIKI IS THE 7395 05:22:47,240 --> 05:22:48,880 THING TO GOOGLE THAT COMES UP 7396 05:22:48,880 --> 05:22:50,720 NUMBER ONE IN SEARCH RESULTS. RESULTS. 7397 05:22:50,720 --> 05:22:56,160 WE HAVE TWO PRIMARY 7398 05:22:56,160 --> 05:22:57,440 COMPREHENSIVE ANALYSIS THAT WE 7399 05:22:57,440 --> 05:23:00,120 INTENT TIME ON, SERUM IS THE 7400 05:23:00,120 --> 05:23:02,760 FIRST FIRST, A GOOD SUBSTRATE AT 7401 05:23:02,760 --> 05:23:06,000 GLYCOPEPTIDE ABUNDANCE AND HEP 7402 05:23:06,000 --> 05:23:09,080 293 CELLS. WE HAVE TARGETED 7403 05:23:09,080 --> 05:23:12,360 ANALYSES WHERE SPIKE PROTEIN 7404 05:23:12,360 --> 05:23:14,160 FROM COVID AND HUMAN ACE TWO 7405 05:23:14,160 --> 05:23:17,040 PROTEIN ANALYZE DIRECTLY IN 7406 05:23:17,040 --> 05:23:18,160 ORDER TO LOOK AT THEIR 7407 05:23:18,160 --> 05:23:19,840 GLYCOSYLATION. NOT SO MUCH A 7408 05:23:19,840 --> 05:23:21,120 COMPREHENSIVE NUMBER OF PROTEINS 7409 05:23:21,120 --> 05:23:25,360 KIND OF ASSAY BUT AS YOU CAN SEE 7410 05:23:25,360 --> 05:23:28,960 NUMBER OF WHITE LANDS SERVE TWO 7411 05:23:28,960 --> 05:23:29,800 SPECIFIC PROTEINS WAS QUITE 7412 05:23:29,800 --> 05:23:38,280 HIGH. LASTLY, WE HAVE THE NIST 7413 05:23:38,280 --> 05:23:39,560 LIBRARY AND WITH THEIR 7414 05:23:39,560 --> 05:23:42,760 PERMISSION ADDED EACH INTO THE 7415 05:23:42,760 --> 05:23:44,920 DATABASE FOR OUR REFERENCE. 7416 05:23:44,920 --> 05:23:49,720 THESE GLYCOPEPTIDES ARE STUDIED 7417 05:23:49,720 --> 05:23:53,760 FIRST USING TYPICAL 7418 05:23:53,760 --> 05:23:55,240 FRAGMENTATION ANALYSIS WHERE WE 7419 05:23:55,240 --> 05:23:58,440 LOOK AT EDD HCC SPECTRA AND 7420 05:23:58,440 --> 05:23:59,840 TRIGGER HCC SPECTRA AFTERWARDS. 7421 05:23:59,840 --> 05:24:02,160 THE PAIR OF SPECTRA REPRESENT 7422 05:24:02,160 --> 05:24:04,440 COMPLIMENTARY INFORMATION WHICH 7423 05:24:04,440 --> 05:24:06,520 TOGETHER GIVE MORE CONVERSATION 7424 05:24:06,520 --> 05:24:09,680 IN THE CORRECTNESS OF 7425 05:24:09,680 --> 05:24:12,320 IDENTIFICATION. THIS IS 7426 05:24:12,320 --> 05:24:14,360 EXTENSIVELY ORGANIZED SET OF 7427 05:24:14,360 --> 05:24:15,240 INFORMATION, HERE IS SAME 7428 05:24:15,240 --> 05:24:21,760 INFORMATION I TOLD YOU ABOUT. 7429 05:24:21,760 --> 05:24:24,680 LESS REGULATION THAT OF SAMPLES 7430 05:24:24,680 --> 05:24:27,120 WITH DIAND AND GLYCOPEPTIDES ARE 7431 05:24:27,120 --> 05:24:29,480 OBSERVED USING DIA ANALYSIS AN 7432 05:24:29,480 --> 05:24:32,360 TRANSITIONS AND THEIR EXTRACTED 7433 05:24:32,360 --> 05:24:33,400 CHROMATOGRAMS ARE AVAILABLE IN 7434 05:24:33,400 --> 05:24:39,680 THIS DATABASE FOR VIEWING. I 7435 05:24:39,680 --> 05:24:42,880 CAN'T CLAIM GPT WIKI SET THE 7436 05:24:42,880 --> 05:24:44,440 WORLD ON FIRE, WE ARE NOT 7437 05:24:44,440 --> 05:24:46,320 PULLING IN THOUSANDS OF VISITORS 7438 05:24:46,320 --> 05:24:48,520 A DAY BUT STEADY USE OF TWO 7439 05:24:48,520 --> 05:24:49,680 YEARS, WE HAVE THE BASELINE 7440 05:24:49,680 --> 05:24:52,120 LEVEL OF USE, WHERE PEOPLE ARE 7441 05:24:52,120 --> 05:24:54,560 VISITING THE SITE AND USING IT 7442 05:24:54,560 --> 05:25:01,080 TO UNDERSTAND WHAT IS GOING ON. 7443 05:25:01,080 --> 05:25:02,960 EXTENSIVE USE AROUND THE WORLD 7444 05:25:02,960 --> 05:25:04,280 THOUGH U.S. IS DOMINANT SITE OF 7445 05:25:04,280 --> 05:25:12,120 USE. THE GLYCOPEPTIDES AND 7446 05:25:12,120 --> 05:25:14,000 GLYCANS AS A SPECIFIC SITE ON 7447 05:25:14,000 --> 05:25:17,560 PROTEIN HAVE BEEN INTEGRATE WITH 7448 05:25:17,560 --> 05:25:19,960 THE GLYCAN INFRASTRUCTURE. YOU 7449 05:25:19,960 --> 05:25:23,000 CAN SEE ANY OF GPT WIKI EVIDENCE 7450 05:25:23,000 --> 05:25:28,960 WE HAVE, DISPLAYED IN GLYGEN AND 7451 05:25:28,960 --> 05:25:30,600 OTHER SOURCES OF EVIDENCE 7452 05:25:30,600 --> 05:25:37,640 PROVIDED FOR GLYCANS ON SITES IN 7453 05:25:37,640 --> 05:25:38,200 GLYCOGEN THESE LINKS PROVIDE 7454 05:25:38,200 --> 05:25:40,560 ACCESS TO THE SPECTRA WHICH 7455 05:25:40,560 --> 05:25:41,960 LABELED SPECTRA AT THAT, WHICH 7456 05:25:41,960 --> 05:25:45,160 GIVE YOU THE EVIDENCE FOR WHY WE 7457 05:25:45,160 --> 05:25:46,680 BELIEVE THAT GLYCAN IS THERE ON 7458 05:25:46,680 --> 05:25:49,920 THAT PROTEIN. I WANT TO GIVE 7459 05:25:49,920 --> 05:25:53,800 SOME IDEA HOW THE DATA ORGANIZED 7460 05:25:53,800 --> 05:25:54,400 AFT HOW YOU CAN FIND WHAT IS 7461 05:25:54,400 --> 05:25:59,160 THERE. WE HAVE A PAGE FOR EACH 7462 05:25:59,160 --> 05:26:01,080 PROTEIN WHICH INDICATES SAMPLES 7463 05:26:01,080 --> 05:26:04,600 THAT IS OBSERVED ON AND SITES 7464 05:26:04,600 --> 05:26:06,120 THAT HAVE GLYCOPEPTIDES ATTACHED 7465 05:26:06,120 --> 05:26:10,160 AS WE CAN SEE THE ASPARAGINE 131 7466 05:26:10,160 --> 05:26:12,840 IS BY FAR MOST NUMBER OF 7467 05:26:12,840 --> 05:26:15,680 GLYCOPEPTIDES OBSERVED. AND 7468 05:26:15,680 --> 05:26:18,680 WHEREAS THE OTHER TWO SITES ARE 7469 05:26:18,680 --> 05:26:20,560 LESS ABUNDANT OR LESS VARIETY OF 7470 05:26:20,560 --> 05:26:25,560 GLYCOPEPTIDES OBSERVED. 7471 05:26:25,560 --> 05:26:28,120 INDIVIDUAL TRANSITION GROUP 7472 05:26:28,120 --> 05:26:29,680 PAGESES INDICATE EVIDENCE FOR A 7473 05:26:29,680 --> 05:26:30,440 SPECIFIC GLYCOPEPTIDE AT SPOT ON 7474 05:26:30,440 --> 05:26:36,960 PROTEIN. WE ARE IDENTIFYING 7475 05:26:36,960 --> 05:26:38,040 THINGS AT LEVEL OF COMPOSITION 7476 05:26:38,040 --> 05:26:40,040 BUT IN ANALYSIS PIPELINES TO 7477 05:26:40,040 --> 05:26:41,200 PROVE OUT AND COLLECT EVIDENCE 7478 05:26:41,200 --> 05:26:44,920 FOR GLYCANS WE ARE ANALYZING 7479 05:26:44,920 --> 05:26:47,160 MULTIPLE STRUCTURE EACH WHICH 7480 05:26:47,160 --> 05:26:48,560 HAVE COMPOSITION SO WE CAN 7481 05:26:48,560 --> 05:26:51,760 ACCOUNT FRAGMENTS SPECIFIC TO 7482 05:26:51,760 --> 05:26:53,560 PARTICULAR TOPOLOGY VERSUS THOSE 7483 05:26:53,560 --> 05:26:57,280 THAT ARE COMMON TO ALL STRUCTURE 7484 05:26:57,280 --> 05:27:00,720 THAT HAVE THAT COMPOSITION. 7485 05:27:00,720 --> 05:27:02,040 CHROMATOGRAM FOR THIS 7486 05:27:02,040 --> 05:27:03,360 GLYCOPEPTIDE, WHICH AGAIN IS 7487 05:27:03,360 --> 05:27:04,960 HELPFUL IN UNDERSTANDING THE 7488 05:27:04,960 --> 05:27:05,960 STRENGTH OF EVIDENCE AND WE 7489 05:27:05,960 --> 05:27:09,680 PRESENT THE PAIR OF SPECTRA HCD 7490 05:27:09,680 --> 05:27:12,360 AT THE TOP AND HCD WHICH 7491 05:27:12,360 --> 05:27:14,480 TOGETHER PROVIDE COMPLIMENTARY 7492 05:27:14,480 --> 05:27:16,160 INFORMATION FOR THE IDENTITY OF 7493 05:27:16,160 --> 05:27:21,840 THIS GLYCOPEPTIDE. IN 7494 05:27:21,840 --> 05:27:25,160 PARTICULAR THIS VIEWER IS 7495 05:27:25,160 --> 05:27:26,560 ZOOMABLE, YOU CAN ZOOM IN ON ANY 7496 05:27:26,560 --> 05:27:28,720 ASPECT OF THE SPECTRA, 7497 05:27:28,720 --> 05:27:30,800 PARTICULARLY USEFUL FOR STUDYING 7498 05:27:30,800 --> 05:27:31,640 THIS FRAGMENT HERE WHICH 7499 05:27:31,640 --> 05:27:35,360 PROVIDES EVIDENCE OF THE CORE 7500 05:27:35,360 --> 05:27:36,080 FUCOSYLATION ON THIS 7501 05:27:36,080 --> 05:27:38,480 GLYCOPEPTIDE. IMPORTANTLY I 7502 05:27:38,480 --> 05:27:41,920 WILL TALK ABOUT THIS IN A 7503 05:27:41,920 --> 05:27:45,720 MINUTE. THIS CHARGE REDUCED 7504 05:27:45,720 --> 05:27:47,600 PREPORKER IN ET ACD SPECTRUM IS 7505 05:27:47,600 --> 05:27:50,240 IMPORTANT FOR US UNDERSTANDING 7506 05:27:50,240 --> 05:27:51,320 WHETHER OR NOT WE HAVE CORRECT 7507 05:27:51,320 --> 05:27:53,520 ASSIGNMENT OF THE PRE-CURSOR FOR 7508 05:27:53,520 --> 05:27:58,000 THESE SPECTRA. HERE IS AN 7509 05:27:58,000 --> 05:28:00,680 EXAMPLE OF THE DIA ANALYSIS OF 7510 05:28:00,680 --> 05:28:03,720 THIS GLYCOPEPTIDE AS PART OF A 7511 05:28:03,720 --> 05:28:04,280 COMPREHENSIVE ANALYSIS FROM 7512 05:28:04,280 --> 05:28:11,000 SERUM. THIS IS DI ANALYSIS 7513 05:28:11,000 --> 05:28:12,760 CONDUCTED USING THE TRANSITION 7514 05:28:12,760 --> 05:28:17,760 LIBRARY FORMED FROM THE PEPTIDE 7515 05:28:17,760 --> 05:28:18,880 FRAGMENTATION SPECTRA. SOMETHING 7516 05:28:18,880 --> 05:28:20,160 I MEANT TO SAY BACK HERE IS PART 7517 05:28:20,160 --> 05:28:21,760 OF THE REASON WE COLLECT TWO 7518 05:28:21,760 --> 05:28:23,680 TYPES OF SPECTRA, THE FIRST 7519 05:28:23,680 --> 05:28:24,640 SPECTRUM IS IMPORTANT BECAUSE IT 7520 05:28:24,640 --> 05:28:26,320 HELPS US IDENTIFY THE 7521 05:28:26,320 --> 05:28:29,240 GLYCOPEPTIDE SUBSTRATE. AND THE 7522 05:28:29,240 --> 05:28:32,960 GLYCAN ITSELF, SECOND SPECTRUM 7523 05:28:32,960 --> 05:28:34,520 IS IMPORTANT BECAUSE HCD 7524 05:28:34,520 --> 05:28:40,760 SPECTRUM IS ONE WE WILL THEN USE 7525 05:28:40,760 --> 05:28:42,240 TO LOOK AT INTENSITY OF 7526 05:28:42,240 --> 05:28:48,760 FRAGMENTS IN THE DIA ANALYSIS. 7527 05:28:48,760 --> 05:28:52,520 WE OBSERVE IN PARTICULAR IN THIS 7528 05:28:52,520 --> 05:28:55,040 SO CALLED SOFT FRAGMENTATION 7529 05:28:55,040 --> 05:28:57,000 MODE, WE LOSE THE DOMINANT 7530 05:28:57,000 --> 05:28:59,240 FRAGMENT HERE THIS BLUE PEAK 7531 05:28:59,240 --> 05:29:03,840 CORRESPONDS TO WHAT WE CALL THE 7532 05:29:03,840 --> 05:29:08,400 YM FROM THE GLYCAN STRUCTURES. 7533 05:29:08,400 --> 05:29:09,720 WE EXPECT THIS FRAGMENT TO 7534 05:29:09,720 --> 05:29:12,560 DOMINATE AND IN DOING SO GIVES 7535 05:29:12,560 --> 05:29:14,120 PARTIAL INFORMATION ABOUT 7536 05:29:14,120 --> 05:29:16,400 STRUCTURE. WE NOTICE THERE'S 7537 05:29:16,400 --> 05:29:20,560 THIS MIDDLE STRUCTURE HERE THAT 7538 05:29:20,560 --> 05:29:25,000 CANNOT FORM CORRECT MASS AS 7539 05:29:25,000 --> 05:29:25,720 TRANSITION, BY OBSERVING THE 7540 05:29:25,720 --> 05:29:27,840 BLUE TRANSITION HERE, SO 7541 05:29:27,840 --> 05:29:29,600 ABUNDANTLY SOME INFORMATION WE 7542 05:29:29,600 --> 05:29:32,560 DON'T PROVE THIS IS THE THE CORE 7543 05:29:32,560 --> 05:29:34,360 FUCOSYLATION FORM BUT THESE 7544 05:29:34,360 --> 05:29:37,960 FRAGMENTS HERE WITH THE ASTERISK 7545 05:29:37,960 --> 05:29:39,680 INDICATE TRANSITIONS SPECIFIC TO 7546 05:29:39,680 --> 05:29:41,360 THE CORE FUCOSYLATED FORM OF THE 7547 05:29:41,360 --> 05:29:50,720 GLYCAN. I WOULD LIKE TO GIVE A 7548 05:29:50,720 --> 05:29:53,160 SENSE OF THE DIFFICULTIES 7549 05:29:53,160 --> 05:29:55,480 INVOLVED IN DOING THIS TYPE OF 7550 05:29:55,480 --> 05:29:58,560 WORK SUCCESSFULLY. THERE WAS 7551 05:29:58,560 --> 05:29:59,760 AWFUL LOT OF WORK THAT WENT INTO 7552 05:29:59,760 --> 05:30:01,920 THIS TO TRY TO COME UP WITH A 7553 05:30:01,920 --> 05:30:05,760 CONCEPT OF NORMALIZED RETENTION 7554 05:30:05,760 --> 05:30:07,840 TIME. AS RELATE TO THESE DIA 7555 05:30:07,840 --> 05:30:09,360 STRATEGY AND GLYCOPEPTIDE 7556 05:30:09,360 --> 05:30:11,000 ANALYSIS. WE TRIED USING THE 7557 05:30:11,000 --> 05:30:12,600 TRADITIONAL IRT PEPTIDES THAT 7558 05:30:12,600 --> 05:30:15,800 ARE COMMERCIALLY AVAILABLE FOR 7559 05:30:15,800 --> 05:30:16,960 GLYCOPROTEOMICS AND WE FOUND 7560 05:30:16,960 --> 05:30:19,480 THEY DON'T DISTRIBUTE WELL OVER 7561 05:30:19,480 --> 05:30:21,360 REGION GLYCOPEPTIDES DISTRIBUTED 7562 05:30:21,360 --> 05:30:22,760 OVER. BUT THEY FORM THEIR 7563 05:30:22,760 --> 05:30:26,080 INITIAL STRATEGY. OUR SECOND 7564 05:30:26,080 --> 05:30:29,960 WAS TO LOOK FOR GLYCOPEPTIDES 7565 05:30:29,960 --> 05:30:31,440 ABUNDANTLY OBSERVED AND WHERE 7566 05:30:31,440 --> 05:30:33,120 SPACE REASONED REASONABLY ACROSS 7567 05:30:33,120 --> 05:30:34,560 THE RETENTION TIME FROM FILE. 7568 05:30:34,560 --> 05:30:36,960 WE USE SELECTED COMMON ABUNDANT 7569 05:30:36,960 --> 05:30:39,160 GLYCOPEPTIDES AS NEXT ATTEMPT 7570 05:30:39,160 --> 05:30:44,520 ONE WE HAVE THEM TO NORMALIZE 7571 05:30:44,520 --> 05:30:45,720 DEAFENINGS TIME. TROUBLE IS 7572 05:30:45,720 --> 05:30:47,080 THOUGH IF YOU PROCEED WITH 7573 05:30:47,080 --> 05:30:48,240 FRACTIONATION APPROACH OR ANY 7574 05:30:48,240 --> 05:30:50,640 OTHER APPROACH TO SELECTIVELY 7575 05:30:50,640 --> 05:30:52,840 TARGET GLYCOPROTEINS. OR SPREAD 7576 05:30:52,840 --> 05:30:55,360 OUT OR MAKE THEM SUCH YOU CAN 7577 05:30:55,360 --> 05:30:59,560 ANALYZE MORE DEEPLY, YOU NO 7578 05:30:59,560 --> 05:31:01,600 LONGER HAVE ACCESS TO COMMON 7579 05:31:01,600 --> 05:31:04,440 ABUNDANT GLYCOPEPTIDES. SO AS 7580 05:31:04,440 --> 05:31:06,960 WE EXPLORED WAYS TO LOOK DEEPER 7581 05:31:06,960 --> 05:31:11,480 INTO THE GLYCOPEPTIDE PROFILE WE 7582 05:31:11,480 --> 05:31:12,080 DISCOVERED GLYCOPEPTIDES NO 7583 05:31:12,080 --> 05:31:15,160 LONGER AVAILABLE AND DIDN'T 7584 05:31:15,160 --> 05:31:17,160 ENCOMPASS THE ENTIRE RETENTION 7585 05:31:17,160 --> 05:31:19,200 TIME RANGE. WE HAD ENOUGH 7586 05:31:19,200 --> 05:31:20,880 GLYCOPEPTIDES WITH NORMALIZED 7587 05:31:20,880 --> 05:31:22,840 PRETENSION TIMES THAT WE CAN USE 7588 05:31:22,840 --> 05:31:25,440 WHATEVER WE CAN FIND. THIS 7589 05:31:25,440 --> 05:31:28,960 WORKS WELL, WE DO INITIAL DIA 7590 05:31:28,960 --> 05:31:30,880 ANALYSIS, WE DABBLE RETENTION 7591 05:31:30,880 --> 05:31:32,240 TIME CONSTRAINT WE TAKE 7592 05:31:32,240 --> 05:31:35,160 CONFIDENTLY IDENTIFIED 7593 05:31:35,160 --> 05:31:37,000 TRANSITIONS AND USE THEM TO 7594 05:31:37,000 --> 05:31:38,640 CREATE CORRECTION FACTOR FOR THE 7595 05:31:38,640 --> 05:31:40,880 NORMALIZE RETENTION TIME. 7596 05:31:40,880 --> 05:31:42,200 NORMALIZE RETENTION TIME MODEL 7597 05:31:42,200 --> 05:31:45,560 IS BUILT FROM THE DDA ANALYSIS 7598 05:31:45,560 --> 05:31:46,800 USING THIS REGRESSION MODEL, 7599 05:31:46,800 --> 05:31:50,680 THIS WORKED QUITE WELL, SIALIC 7600 05:31:50,680 --> 05:31:54,000 ACID IS IMPORTANT WE DISCOVERED 7601 05:31:54,000 --> 05:31:55,480 ME THIONEINE OXIDATION MAKES A 7602 05:31:55,480 --> 05:31:56,560 DIFFERENCE AND THE 7603 05:31:56,560 --> 05:31:57,560 MONOSACCHARIDES IN STRUCTURE. WE 7604 05:31:57,560 --> 05:31:58,920 BUILT A SECOND REGRESSION MODEL 7605 05:31:58,920 --> 05:32:00,720 FOR EACH PEPTIDE. FOR EACH 7606 05:32:00,720 --> 05:32:05,600 UNDERLYING PEPTIDE SU SUBSTRATE. 7607 05:32:05,600 --> 05:32:07,720 THIS WAS GOOD AND BAD. THE 7608 05:32:07,720 --> 05:32:11,400 PEPTIDE ITSELF IS THE PRIMARY 7609 05:32:11,400 --> 05:32:12,280 DETERMINANT OF THE RETENTION 7610 05:32:12,280 --> 05:32:15,640 TIME FOR CAT. WE NEED THESE 7611 05:32:15,640 --> 05:32:17,800 OTHER FACTORS IN THERE THERE. 7612 05:32:17,800 --> 05:32:19,200 UNFORTUNATELY IF WE DON'T 7613 05:32:19,200 --> 05:32:20,360 OBSERVE DIFFERENT FORMS OF 7614 05:32:20,360 --> 05:32:21,160 GLYCOPEPTIDES FOR UNDERLYING 7615 05:32:21,160 --> 05:32:23,840 PEPTIDE SUBSTRATE, WHICH DON'T 7616 05:32:23,840 --> 05:32:25,080 HAVE ENOUGH INFORMATION TO FIT 7617 05:32:25,080 --> 05:32:27,960 THIS MODEL PARTICULARLY WELL. 7618 05:32:27,960 --> 05:32:30,560 WE WORKED HARD TO USE EVERY TOOL 7619 05:32:30,560 --> 05:32:33,880 AT OUR DISPOSAL TO MAKE THIS 7620 05:32:33,880 --> 05:32:35,760 GLYCOPEPTIDE ID SUBSTRATE HIGH 7621 05:32:35,760 --> 05:32:37,640 QUALITY. WE CURATED A LIMITED 7622 05:32:37,640 --> 05:32:40,880 SET OF PROTEINS AND GLYCANS, WE 7623 05:32:40,880 --> 05:32:43,840 USED FDR FILTERING AT EVERY 7624 05:32:43,840 --> 05:32:46,720 PLACE IN ORDER TO TRY AND ENSURE 7625 05:32:46,720 --> 05:32:48,400 WE DIDN'T HAVE FALSE POSITIVES. 7626 05:32:48,400 --> 05:32:50,200 WE USE NORMALIZE RETENTION TIME 7627 05:32:50,200 --> 05:32:52,160 FIT TO MAKE SURE THAT THE IDs 7628 05:32:52,160 --> 05:32:54,520 WHERE WE EXPECTED THEM TO BE IN 7629 05:32:54,520 --> 05:32:55,680 TERMS OF THE PREVIOUS 7630 05:32:55,680 --> 05:32:57,160 IDENTIFICATION WE SAW. AND WE 7631 05:32:57,160 --> 05:33:01,680 CHECKED THINGS LIKE THE 7632 05:33:01,680 --> 05:33:03,760 RETENTION TIME TO MAKE SURE IT 7633 05:33:03,760 --> 05:33:06,800 HAD REASONABLE SHAPE. ALL THESE 7634 05:33:06,800 --> 05:33:10,360 THINGS WERE GOOD ENSURING WE HAD 7635 05:33:10,360 --> 05:33:12,000 HIGH QUALITY GLYCOPEPTIDE IDs 7636 05:33:12,000 --> 05:33:13,560 BUT ULTIMATELY DISCOVERED THAT 7637 05:33:13,560 --> 05:33:17,480 WAS NOT ENOUGH. BECAUSE THERE 7638 05:33:17,480 --> 05:33:22,600 ARE PERFECTLY MANY THINGS, MANY 7639 05:33:22,600 --> 05:33:25,040 GLYCOPEPTIDES WHICH 7640 05:33:25,040 --> 05:33:28,520 SIGNIFICANTLY AND IMPORTANTLY 7641 05:33:28,520 --> 05:33:29,160 NOT RANDOM BUT NEVERTHELESS 7642 05:33:29,160 --> 05:33:32,520 WRONG. WE FOUND THIS BECAUSE WE 7643 05:33:32,520 --> 05:33:33,760 STARTED USING TWO DIFFERENT 7644 05:33:33,760 --> 05:33:35,720 SEARCH ENGINES AND THEY GAVE 7645 05:33:35,720 --> 05:33:36,600 INCONSISTENT RESULTS FOR THE 7646 05:33:36,600 --> 05:33:38,640 SAME SPECTRUM. AND IT WAS REALLY 7647 05:33:38,640 --> 05:33:42,400 THIS PROBLEM, THE TWO 7648 05:33:42,400 --> 05:33:44,280 PRE-CURSORS BEING ONE SIALIC 7649 05:33:44,280 --> 05:33:45,120 ACID THAT KILLS YOU. BECAUSE YOU 7650 05:33:45,120 --> 05:33:46,560 CAN'T -- THERE IS NOTHING IN 7651 05:33:46,560 --> 05:33:48,920 THERE TO FIND A WAY TO RESOLVE 7652 05:33:48,920 --> 05:33:50,520 THIS IN MOST SEARCH ENGINES WE 7653 05:33:50,520 --> 05:33:54,920 TRIED. SO WHAT WE DISCOVERED IS 7654 05:33:54,920 --> 05:33:58,080 HELPFUL TO TAKE CARE OF THIS 7655 05:33:58,080 --> 05:34:00,560 PROBLEM. IT TYPICALLY HAS A 7656 05:34:00,560 --> 05:34:02,280 CHARGE REDUCED PRE-CURSOR 7657 05:34:02,280 --> 05:34:06,440 ISOTOPE CLUSTER. YOU CAN USE 7658 05:34:06,440 --> 05:34:08,000 THIS TO CHECK PRE-CURSOR FOR 7659 05:34:08,000 --> 05:34:10,040 IDENTIFICATION YOU HAVE, FITS ON 7660 05:34:10,040 --> 05:34:12,360 THAT MONOISOTOPPIC PEAK OF THAT 7661 05:34:12,360 --> 05:34:15,240 PRECURSOR ISOTOPE CLUSTER. IF 7662 05:34:15,240 --> 05:34:17,840 YOU HAVE THE INSTITUTION OF TWO 7663 05:34:17,840 --> 05:34:19,960 FUCOSE INSTEAD, IT SHIPS OVER 7664 05:34:19,960 --> 05:34:23,000 ONE AND NOT IN THE RIGHT SPOT. 7665 05:34:23,000 --> 05:34:24,960 IN OUR ANALYSIS, WHERE WE WERE 7666 05:34:24,960 --> 05:34:27,440 LOOKING HARD FOR GLYCOPEPTIDES, 7667 05:34:27,440 --> 05:34:29,240 WE FOUND N OF ONE SPECTRUM 7668 05:34:29,240 --> 05:34:31,280 DIDN'T HAVE PRECURSOR ISOTOPE 7669 05:34:31,280 --> 05:34:34,160 CLUSTER TO RECOVER, ONLY THE 7670 05:34:34,160 --> 05:34:35,720 FRAGMENTATION SPECTRA IN ETD 7671 05:34:35,720 --> 05:34:38,880 DID. WE ALSO REQUIRE IF WE GOT A 7672 05:34:38,880 --> 05:34:40,360 CLAIM OF FUCOSE PEPTIDE AN 7673 05:34:40,360 --> 05:34:43,600 GLYCOPEPTIDE IT DOESN'T HAVE YES 7674 05:34:43,600 --> 05:34:45,040 ION WHICH SUPPORTS THAT BLAME. 7675 05:34:45,040 --> 05:34:48,120 AFTER DOING THESE STEPS, ALMOST 7676 05:34:48,120 --> 05:34:52,200 ALL OF THE 2, 3, 4 FUCOSE FORMS 7677 05:34:52,200 --> 05:34:55,160 IN OUR DATABASE WERE ELIMINATED. 7678 05:34:55,160 --> 05:34:57,240 WE HAVE SIGNIFICANT DISJUST OF 7679 05:34:57,240 --> 05:35:01,920 ALL MULTIPLY FUCOSE INFORMED -- 7680 05:35:01,920 --> 05:35:04,160 FUCOSYLATED FORMS THAT I OBSERVE 7681 05:35:04,160 --> 05:35:05,040 IN VARIOUS ANNOTATION 7682 05:35:05,040 --> 05:35:08,560 SUBSTRATES. HAPPY TO TALK MORE 7683 05:35:08,560 --> 05:35:13,680 IF YOU HAVE QUESTIONS. THIS IS 7684 05:35:13,680 --> 05:35:15,600 UP AND WE ARE HOPING THAT IT IS 7685 05:35:15,600 --> 05:35:18,520 USEFUL TO THE COMMUNITY. IF YOU 7686 05:35:18,520 --> 05:35:19,560 HAVE A DATA SET THAT YOU THINK 7687 05:35:19,560 --> 05:35:21,320 MIGHT BE WELL DISPLAYED IN THIS 7688 05:35:21,320 --> 05:35:23,960 SETTING, IT HAS A CLEAR PATH TO 7689 05:35:23,960 --> 05:35:27,840 GET INTO GLYGEN AS ANNATIONS 7690 05:35:27,840 --> 05:35:29,920 WITH SUPPORTING SECOND -- 7691 05:35:29,920 --> 05:35:30,760 ANNOTATIONS WITH SUPPORTING 7692 05:35:30,760 --> 05:35:33,480 SPECTRASPECTRA. HAPPY TO TALK TO YOU TO 7693 05:35:33,480 --> 05:35:34,880 FIGURE HOW TO GET INTO THAT TA 7694 05:35:34,880 --> 05:35:55,560 INFRASTRUCTURE. THANK YOU. 7695 05:35:55,560 --> 05:36:23,520 (OFF MIC) 7696 05:36:23,520 --> 05:36:34,680 (NO AUDIO) UO 1 GRANT FOR ALL OF 7697 05:36:34,680 --> 05:36:37,480 YOU IN PERSON, I FIRST WANT TO 7698 05:36:37,480 --> 05:36:40,680 THANK CARL PAM AND AMANDA AND 7699 05:36:40,680 --> 05:36:44,560 DOUG FOR THE LEADERSHIP AND 7700 05:36:44,560 --> 05:36:45,800 SUPPORT FOR THIS PARTICULAR 7701 05:36:45,800 --> 05:36:49,280 PROGRAM. SO AS MANY OF YOU KNOW, 7702 05:36:49,280 --> 05:36:50,680 WE HAVE BEEN DOING AND SPENDING 7703 05:36:50,680 --> 05:36:54,760 QUITE A BIT OF EFFORT TO DEVELOP 7704 05:36:54,760 --> 05:36:56,560 NEW TOOLS FOR FACILITATING HIGH 7705 05:36:56,560 --> 05:36:57,840 THROUGH PUT GLYCAN AND 7706 05:36:57,840 --> 05:37:01,000 GLYCOPEPTIDE QUANTITATION. FOR 7707 05:37:01,000 --> 05:37:02,480 THIS AUDIENCE PERHAPS NOT SO 7708 05:37:02,480 --> 05:37:04,600 NECESSARY TO REALLY INTRODUCE 7709 05:37:04,600 --> 05:37:06,760 THE IMPORTANCE OF GLYCOSYLATION 7710 05:37:06,760 --> 05:37:10,560 WHICH IS AMONG MOST IMPORTANT 7711 05:37:10,560 --> 05:37:11,880 COMPACT CHEMICAL MODIFICATION 7712 05:37:11,880 --> 05:37:14,160 FOR PROTEINS AND THIS CAN CONFER 7713 05:37:14,160 --> 05:37:16,240 GREAT VARIETY OF DIFFERENT 7714 05:37:16,240 --> 05:37:17,960 FUNCTIONALITY RANGING FROM CELL 7715 05:37:17,960 --> 05:37:24,080 CELL COMMUNICATION, 7716 05:37:24,080 --> 05:37:25,920 IMMUNORECOGNITION AND ABERRANT 7717 05:37:25,920 --> 05:37:27,640 EXPRESSION OF MANY DISEASE 7718 05:37:27,640 --> 05:37:29,360 GLYCAN EXPRESSION IS IMPLICATED 7719 05:37:29,360 --> 05:37:32,720 IN A VARIETY OF DISEASE, RANGING 7720 05:37:32,720 --> 05:37:34,160 FROM CANCER, CARDIOVASCULAR 7721 05:37:34,160 --> 05:37:35,360 DISORDERS AND NEURODEGENERATIVE 7722 05:37:35,360 --> 05:37:39,680 DISEASE. CONSEQUENTLY IT IS 7723 05:37:39,680 --> 05:37:42,760 IMPORTANT TO QUANTIFY GLYCANS IN 7724 05:37:42,760 --> 05:37:43,920 VARIETY OF BIOLOGICAL SYSTEMS. 7725 05:37:43,920 --> 05:37:46,160 SO IN OUR RECENT REVIEW ARTICLES 7726 05:37:46,160 --> 05:37:49,880 WE HAVE SURVEYED A NUMBER OF 7727 05:37:49,880 --> 05:37:52,960 CURRENT STRATEGIES TO QUANTIFY 7728 05:37:52,960 --> 05:37:55,200 GLYCOME AND GLYCOPEPTIDES AT THE 7729 05:37:55,200 --> 05:37:58,000 FREELY RELEASE GLYCAN LEVEL OR 7730 05:37:58,000 --> 05:38:01,520 INTACT GLYCOPEPTIDE LEVEL. 7731 05:38:01,520 --> 05:38:04,040 VARIETY OF WAYS TO DO THIS FOR 7732 05:38:04,040 --> 05:38:05,400 QUANTITATION LABEL FREE OR 7733 05:38:05,400 --> 05:38:10,920 METABOLIC LABELING WITH ISOTOPES 7734 05:38:10,920 --> 05:38:14,320 OR ISOTOPES IN CHEMICAL TACT AND 7735 05:38:14,320 --> 05:38:16,040 DONE IN MS 1 STAGE, THIS 7736 05:38:16,040 --> 05:38:16,840 PARTICULAR TARGET FOCUSES MORE 7737 05:38:16,840 --> 05:38:20,040 ON THE MS 2 BASE OR ISOBARRY 7738 05:38:20,040 --> 05:38:27,920 TAGGING STRATEGY. THERE ARE 7739 05:38:27,920 --> 05:38:29,440 SEVERAL LIMITATIONS WITH CURRENT 7740 05:38:29,440 --> 05:38:31,560 STRATEGY. THE METABOLIC LABELING 7741 05:38:31,560 --> 05:38:35,520 IS RELATIVELY EXPENSIVE AND ALSO 7742 05:38:35,520 --> 05:38:39,360 STABLE ISOTOPE LABELING AT THE 7743 05:38:39,360 --> 05:38:43,120 MS 1 STAGE INTRODUCE OVERLAPPING 7744 05:38:43,120 --> 05:38:46,400 ISSUE AND ALSO INTRODUCE MORE 7745 05:38:46,400 --> 05:38:49,680 CHEMICAL COMPLEXITY AND OFTEN 7746 05:38:49,680 --> 05:38:52,560 COME WITH DUPLEX OR TRIPLEX 7747 05:38:52,560 --> 05:38:54,680 VERSION. THIS IN PARTICULAR WHEN 7748 05:38:54,680 --> 05:38:56,520 INCORPORATING DATA DEPENDENT 7749 05:38:56,520 --> 05:38:59,520 ACQUISITION COULD LEAD TO 7750 05:38:59,520 --> 05:39:02,160 DECREASED INSTRUMENT DUTY CYCLE 7751 05:39:02,160 --> 05:39:04,320 WHICH CREATES SACRIFICE DEPTH OF 7752 05:39:04,320 --> 05:39:05,320 QUANTIFICATION AND 7753 05:39:05,320 --> 05:39:07,360 IDENTIFICATION. OBVIOUSLY THE MS 7754 05:39:07,360 --> 05:39:09,800 2 BASE ISOTAGGING STRATEGY CAN 7755 05:39:09,800 --> 05:39:11,400 ADDRESS THESE LIMITATIONS IN ONE 7756 05:39:11,400 --> 05:39:16,560 OF THE COMMERCIAL AVAILABLE 7757 05:39:16,560 --> 05:39:18,760 PRODUCT AMINOOXI TMT IS 7758 05:39:18,760 --> 05:39:19,960 ADDRESSING LIMITATIONS IN TERMS 7759 05:39:19,960 --> 05:39:22,680 OF PROVIDING HIGH THROUGH PUT MS 7760 05:39:22,680 --> 05:39:24,720 2 BASE QUANTIFICATION. HOWEVER 7761 05:39:24,720 --> 05:39:26,280 THERE'S STILL LIMITATIONS THAT I 7762 05:39:26,280 --> 05:39:31,040 WILL ELABORATE IN TERMS OF THE 7763 05:39:31,040 --> 05:39:33,160 REPORTER AND COMPLEXITY FOR 7764 05:39:33,160 --> 05:39:37,520 THESE DIFFERENT GLYCANS SHOWING 7765 05:39:37,520 --> 05:39:39,360 HERE IS ISOBARIC LABELING 7766 05:39:39,360 --> 05:39:44,560 STRATEGY, COMMERCIALLY AVAILABLE 7767 05:39:44,560 --> 05:39:45,640 6 COMPLEX TMT VERSION. WE HAVE 7768 05:39:45,640 --> 05:39:47,960 THESE SET UP REPORTER GROUP AND 7769 05:39:47,960 --> 05:39:49,640 THEN THE BALANCE GROUP SO SUM OF 7770 05:39:49,640 --> 05:39:52,760 THE TWO PORTIONS ARE GIVING RISE 7771 05:39:52,760 --> 05:39:57,680 TO IDENTIFY MASS SO ALLOW US TO 7772 05:39:57,680 --> 05:39:58,680 DIFFERENTIALLY LABEL FOUR SETS 7773 05:39:58,680 --> 05:40:01,480 OF SAMPLES IN THE MS 1 MODE EACH 7774 05:40:01,480 --> 05:40:04,440 GLYCAN WILL SHOW AS SINGLE PEAK, 7775 05:40:04,440 --> 05:40:06,760 WITH SAME SHIFT AND THEN THE 7776 05:40:06,760 --> 05:40:10,240 MSMS WOULD GENERATE THESE 7777 05:40:10,240 --> 05:40:11,680 REPORTER ION ON THE PART FROM 7778 05:40:11,680 --> 05:40:13,320 EACH OTHER ALLOWING RELATIVE 7779 05:40:13,320 --> 05:40:14,280 QUANTIFYCATION. AS WE CAN SEE 7780 05:40:14,280 --> 05:40:18,120 THIS IS THE COMMERCIALLY 7781 05:40:18,120 --> 05:40:20,560 AVAILABLE TMP REAGENT AND 7782 05:40:20,560 --> 05:40:24,200 THERE'S RED STARS INDICATING THE 7783 05:40:24,200 --> 05:40:25,680 POSITION OF THE HEAVY STABLE 7784 05:40:25,680 --> 05:40:28,560 ISOTOPES. HERE SHOWS THE 7785 05:40:28,560 --> 05:40:29,760 ALDEHYDE KETONE REACTIVE GROUP 7786 05:40:29,760 --> 05:40:31,520 THAT WILL REACT WITH REDUCING 7787 05:40:31,520 --> 05:40:35,200 ENDS OF GLYCANS AND MASS 7788 05:40:35,200 --> 05:40:37,680 BALANCER REPORTER. IN THE MSMS 7789 05:40:37,680 --> 05:40:39,240 MODE THIS WILL GENERATE THE 7790 05:40:39,240 --> 05:40:40,880 CYCLIC REPORTER ION THAT ALLOWS 7791 05:40:40,880 --> 05:40:42,360 US TO PERFORM RELATIVE 7792 05:40:42,360 --> 05:40:45,560 QUANTIFICATION. HOWEVER THIS 7793 05:40:45,560 --> 05:40:48,320 APPROACH SUFFERS FROM SOME OF 7794 05:40:48,320 --> 05:40:50,240 THE LIMITATION IN PARTICULAR 7795 05:40:50,240 --> 05:40:52,320 LOOKING AT LOW ABUNDANCE COMPLEX 7796 05:40:52,320 --> 05:40:56,360 N GLYCANS SO GENERATE IN 7797 05:40:56,360 --> 05:40:59,040 PARTICULAR RELATIVELY LOWER 7798 05:40:59,040 --> 05:41:01,960 REPORTER ION YIELD AND LEAD TO 7799 05:41:01,960 --> 05:41:03,160 INACCURATE REPORTER ION RATIO 7800 05:41:03,160 --> 05:41:04,640 AND YOU CAN SEE FROM THIS 7801 05:41:04,640 --> 05:41:07,000 EXPERIMENT THE LOW COLLISIONAL 7802 05:41:07,000 --> 05:41:07,480 ENERGY WE HAVE MINIMUM 7803 05:41:07,480 --> 05:41:10,360 PRODUCTION OF THE REPORTER ION. 7804 05:41:10,360 --> 05:41:11,920 SO IN ORDER TO BOOST THAT 7805 05:41:11,920 --> 05:41:15,480 REPORTER ION WE NEED TO PERFORM 7806 05:41:15,480 --> 05:41:19,080 HIGH ENERGY DISSOCIATION AND 7807 05:41:19,080 --> 05:41:21,680 THIS LEADS TO EXCESSIVE 7808 05:41:21,680 --> 05:41:24,120 FRAGMENTATION SO NO BACKWARD 7809 05:41:24,120 --> 05:41:25,960 FRAGMENTATION DIFFICULTY FOR 7810 05:41:25,960 --> 05:41:26,560 IDENTIFICATION. AS A RESULT 7811 05:41:26,560 --> 05:41:28,400 OFTEN TIMES WE HAVE TO DO BOTH 7812 05:41:28,400 --> 05:41:30,280 LOW ENERGY AND HIGH ENERGY 7813 05:41:30,280 --> 05:41:32,200 COLLISION DISSOCIATION. SO TO 7814 05:41:32,200 --> 05:41:35,520 ADDRESS THIS LIMITATION OUR 7815 05:41:35,520 --> 05:41:39,320 GROUP TALENTED STUDENT HAS TOOK 7816 05:41:39,320 --> 05:41:41,560 ADVANTAGE OF NATURALLY OCCURRING 7817 05:41:41,560 --> 05:41:42,640 IMMUNOASSAY BUILDING BLOCKS AND 7818 05:41:42,640 --> 05:41:48,680 HERE WE SELECTED LEUCINE Z OUR AS OUR 7819 05:41:48,680 --> 05:41:50,640 BUILDING BLOCK AFTER 7820 05:41:50,640 --> 05:41:51,880 DIMETHYLATION IN PRESENCE OF 7821 05:41:51,880 --> 05:41:54,200 GLYCENE METHYL ESTER WE CAN 7822 05:41:54,200 --> 05:41:56,320 INSTALL GLYCENE AS LINKER AND 7823 05:41:56,320 --> 05:42:01,560 REACT TO FORM THIS HYDROSIDE 7824 05:42:01,560 --> 05:42:05,600 REACTIVE GROUP, THIS SUGAR TEST 7825 05:42:05,600 --> 05:42:07,600 LABEL ANY CARBONYL CONTAINING 7826 05:42:07,600 --> 05:42:09,960 COMPOUND. BY INSTALLING 7827 05:42:09,960 --> 05:42:11,600 DIFFERENT STABLE ISOTOPES SUCH 7828 05:42:11,600 --> 05:42:14,880 AS COLOR CODED HERE WE CAN 7829 05:42:14,880 --> 05:42:17,600 CREATE SHOULDER TAG REPORTER ION 7830 05:42:17,600 --> 05:42:20,560 IS ONE APART FROM EACH OTHER 7831 05:42:20,560 --> 05:42:24,800 FROM 115 TO 118. WITH THIS SUGAR 7832 05:42:24,800 --> 05:42:27,360 LABELING STRATEGY WE DEVELOPED 7833 05:42:27,360 --> 05:42:29,880 INTEGRATED WORK FLOW TO LOOK AT 7834 05:42:29,880 --> 05:42:31,120 GLYCOME IDENTIFICATION SO FIRST 7835 05:42:31,120 --> 05:42:34,560 WE WANT TO EVALUATE WITH 7836 05:42:34,560 --> 05:42:38,800 GLYCOPROTEIN BOVINE GLOBULIN 7837 05:42:38,800 --> 05:42:39,600 FREELY RELEASED GLOBULIN 7838 05:42:39,600 --> 05:42:41,480 DENATURED PENGASE AFTER RELEASE 7839 05:42:41,480 --> 05:42:45,960 OF GLYCANS, IN KNOWN RASH YES 7840 05:42:45,960 --> 05:42:47,440 AND THEN LABEL DIFFERENTIALLY 7841 05:42:47,440 --> 05:42:51,120 LABEL WITH 4 COMPLEX SUGAR AND 7842 05:42:51,120 --> 05:42:54,960 MS 1 MODE GENERATE FOR EACH OF 7843 05:42:54,960 --> 05:42:57,080 THE SINGLE PEAK AND MS 2 ALLOWS 7844 05:42:57,080 --> 05:42:58,680 US TO QUANTIFY. SO THE FIRST 7845 05:42:58,680 --> 05:43:00,440 THING WE WANT TO DO IS TO CAM 7846 05:43:00,440 --> 05:43:02,560 PAIR THE LABELING EFFICIENCY 7847 05:43:02,560 --> 05:43:05,760 COMPARED TO THE COMMERCIAL 7848 05:43:05,760 --> 05:43:07,320 PRODUCT. SO HERE WE CAN SEE THAT 7849 05:43:07,320 --> 05:43:12,720 THE TOP IS THE UNLABELED NATIVE 7850 05:43:12,720 --> 05:43:16,560 GLYCANS AND THE AMINO OXTNT AND 7851 05:43:16,560 --> 05:43:18,160 BOTTOM IS SUGAR TAG IN THE 7852 05:43:18,160 --> 05:43:20,400 COMMERCIAL PRODUCT WE STILL HAVE 7853 05:43:20,400 --> 05:43:22,560 50% UNLABELED PRODUCT AND THEN 7854 05:43:22,560 --> 05:43:24,560 WITH OUR SUGAR TAB WE CAN SEE 7855 05:43:24,560 --> 05:43:25,880 FOR THESE NUMEROUS DIFFERENT 7856 05:43:25,880 --> 05:43:29,360 TYPE OF N GLYCANS WE HAVE 100% 7857 05:43:29,360 --> 05:43:32,000 CONVERSION LABELING EFFICIENCY. 7858 05:43:32,000 --> 05:43:33,520 ANOTHER VERY IMPORTANT FACTOR TO 7859 05:43:33,520 --> 05:43:35,040 COMPARE IS TO FRAGMENTATION 7860 05:43:35,040 --> 05:43:37,720 BECAUSE THAT IS WHERE WE BASE 7861 05:43:37,720 --> 05:43:40,240 OUR QUANTIFYCATION. SO HERE WE 7862 05:43:40,240 --> 05:43:42,160 PERFORM SIDE BY SIDE COMPARISON 7863 05:43:42,160 --> 05:43:48,520 WITH THE AMINO OXTNT SUGAR H HCD 7864 05:43:48,520 --> 05:43:49,360 COLLISIONAL ENERGY BETWEEN 25 7865 05:43:49,360 --> 05:43:53,200 AND 30. IN THIS CASE YOU CAN SEE 7866 05:43:53,200 --> 05:43:55,360 THAT IN BOTH CASES SUGAR TAG 7867 05:43:55,360 --> 05:43:56,520 PRODUCED MORE INTENSE REPORTER 7868 05:43:56,520 --> 05:43:59,400 ION INDICATING THAT WE CAN 7869 05:43:59,400 --> 05:44:01,480 ACTUALLY PERFORM RELATIVE 7870 05:44:01,480 --> 05:44:02,920 QUANTIFICATION FOR LOWER 7871 05:44:02,920 --> 05:44:06,720 ABUNDANCE GLYCANS AN THIS WAS 7872 05:44:06,720 --> 05:44:09,280 STILL GENERATE PRETTY ABUNDANT 7873 05:44:09,280 --> 05:44:10,560 FRAGMENTATION FOR 7874 05:44:10,560 --> 05:44:11,960 IDENTIFICATION. SIMILAR 7875 05:44:11,960 --> 05:44:13,520 PERFORMANCE ACHIEVE FOR SIALIC 7876 05:44:13,520 --> 05:44:16,160 ACID IN FUCOSE CONTAINING GLYCAN 7877 05:44:16,160 --> 05:44:21,360 SHOWN IN THIS EXAMPLE. WE LOOK 7878 05:44:21,360 --> 05:44:23,320 AT COLLECTIVE COMPLEX LIKE 7879 05:44:23,320 --> 05:44:25,600 PICTURES WITH KNOWN RATIO 1 TO 1 7880 05:44:25,600 --> 05:44:29,040 TO 5 TO 10. AS YOU CAN SEE FROM 7881 05:44:29,040 --> 05:44:31,920 THIS BOX PLOT WE GET GOOD 7882 05:44:31,920 --> 05:44:33,000 QUANTITATION ACCURACY SHOWING 7883 05:44:33,000 --> 05:44:34,360 THE BOTTOM ARE TWO 7884 05:44:34,360 --> 05:44:37,080 REPRESENTATIVE GLYCAN EXAMPLES, 7885 05:44:37,080 --> 05:44:39,360 BOTH FUCOSE AND SIALIC ACID 7886 05:44:39,360 --> 05:44:42,920 CONTAINING GLYCANS WE GET PRETTY 7887 05:44:42,920 --> 05:44:44,520 GOOD ACCURACY QUANTIFICATION. SO 7888 05:44:44,520 --> 05:44:49,840 NEXT WE WANT TO APPLY SUGAR TAG 7889 05:44:49,840 --> 05:44:50,960 TO COMPLEX REAL BIOLOGICAL 7890 05:44:50,960 --> 05:44:55,280 EXAMPLES SO WE TEAMED WITH CHRIS 7891 05:44:55,280 --> 05:44:56,760 IKONOMIDOU AT MEDICAL SCHOOL IN 7892 05:44:56,760 --> 05:44:58,200 UW MADISON, HER GROUP IS 7893 05:44:58,200 --> 05:45:02,160 INTERESTED IN LOOKING AT THESE 7894 05:45:02,160 --> 05:45:05,240 CHEMOTHERAPY TREATED 7895 05:45:05,240 --> 05:45:08,400 LYMPHOBLASTIC LEUKEMIA CHILDREN 7896 05:45:08,400 --> 05:45:09,640 WHERE WE CAN COLLECT SERUM 7897 05:45:09,640 --> 05:45:12,560 SAMPLES AND LOOK AT THE AFTER 7898 05:45:12,560 --> 05:45:14,800 CERTAIN TIME PERIOD OF 7899 05:45:14,800 --> 05:45:15,720 CHEMOTHERAPY, THE DYNAMIC 7900 05:45:15,720 --> 05:45:16,720 CHANGES OF THESE DIFFERENT 7901 05:45:16,720 --> 05:45:19,400 GLYCANS. SO HERE SHOWS THE 7902 05:45:19,400 --> 05:45:22,960 EXPERIMENTAL SET UPSTART WITH 7903 05:45:22,960 --> 05:45:24,040 BASELINE BEFORE TREATMENT AND 7904 05:45:24,040 --> 05:45:25,520 ONE MONTH AND THREE MONTHS AND 7905 05:45:25,520 --> 05:45:27,840 SIX MONTHS AFTER TREATMENT. SO 7906 05:45:27,840 --> 05:45:31,440 THIS WOULD THEN ALLOW US TO 7907 05:45:31,440 --> 05:45:33,040 UTILIZE FOUR COMPLEX SUGAR TAG 7908 05:45:33,040 --> 05:45:36,400 TO QUANTIFY LARGE NUMBER OF 7909 05:45:36,400 --> 05:45:37,680 GLYCANS COLLECTED FROM SERUM 7910 05:45:37,680 --> 05:45:39,840 SAMPLES. SO FROM THIS BAR GRAPH 7911 05:45:39,840 --> 05:45:42,720 YOU CAN SEE THE Y AXIS IS 7912 05:45:42,720 --> 05:45:43,880 TREATED AFTER CONTROL, AND HERE 7913 05:45:43,880 --> 05:45:46,520 WE ARE LOOKING AT NUMBER OF 7914 05:45:46,520 --> 05:45:48,320 THESE DIFFERENT GLYCANS, ALL 7915 05:45:48,320 --> 05:45:51,400 TOGETHER WE WERE ABLE TO 7916 05:45:51,400 --> 05:45:55,240 QUANTIFY 145 N GLYCANS INCLUDING 7917 05:45:55,240 --> 05:45:56,720 68 N GLYCANS PRESENT IN ALL 7918 05:45:56,720 --> 05:45:58,880 THREE PATIENTS AND THESE WERE 7919 05:45:58,880 --> 05:46:01,560 DONE WITH THE BIOLOGICAL AND 7920 05:46:01,560 --> 05:46:03,240 TECHNICAL TRIPLICATES. SO 7921 05:46:03,240 --> 05:46:05,360 OBVIOUSLY THE NEXT STEP FOR US 7922 05:46:05,360 --> 05:46:07,360 IS TO FURTHER INCREASE THROUGH 7923 05:46:07,360 --> 05:46:10,160 PUT OR MULTIPLEXING CAPACITY 7924 05:46:10,160 --> 05:46:11,680 BECAUSE THERE ARE NUMBER OF 7925 05:46:11,680 --> 05:46:13,920 ADVANTAGES IN THAT WE CAN REDUCE 7926 05:46:13,920 --> 05:46:16,160 AMOUNT OF ANY ONE GIVEN SAMPLE 7927 05:46:16,160 --> 05:46:18,560 AND THIS CAN ALSO REDUCE SAMPLE 7928 05:46:18,560 --> 05:46:20,760 PREPARATION AND ANALYSIS TIME. 7929 05:46:20,760 --> 05:46:22,360 AND MORE IMPORTANTLY, THIS WOULD 7930 05:46:22,360 --> 05:46:24,240 ALLOW US TO LOOK AT LARGER 7931 05:46:24,240 --> 05:46:27,120 NUMBER OF BIOLOGICAL REPLICATES 7932 05:46:27,120 --> 05:46:29,560 OR DIFFERENT CONDITIONS AND 7933 05:46:29,560 --> 05:46:31,960 FINALLY THIS COULD IMPROVE LCMS 7934 05:46:31,960 --> 05:46:33,400 DATA OVERLAP. SO IN ORDER TO 7935 05:46:33,400 --> 05:46:35,920 ACHIEVE THAT, WE HAVE TAKEN 7936 05:46:35,920 --> 05:46:38,360 ADVANTAGE OF THE HIGH RESOLUTION 7937 05:46:38,360 --> 05:46:40,240 INSTRUMENTATION BUT ALSO 7938 05:46:40,240 --> 05:46:42,040 PHENOMENON CALLED MASS DEFECT 7939 05:46:42,040 --> 05:46:47,360 AND THE BASIC IDEA IS THAT SMALL 7940 05:46:47,360 --> 05:46:48,000 NEUTRON BINDING ENERGY 7941 05:46:48,000 --> 05:46:49,880 DIFFERENCE RESULT IN SMALL MASS 7942 05:46:49,880 --> 05:46:54,080 DIFFERENCE, IF WE SWAP WITH 7943 05:46:54,080 --> 05:47:00,240 CARBON 13 THIS REEDS TO 2.9 7944 05:47:00,240 --> 05:47:00,960 MILIDALTON MASS DIFFERENCE, NOT 7945 05:47:00,960 --> 05:47:02,000 DIFFERENTIATED WITH HIGH 7946 05:47:02,000 --> 05:47:02,680 RESOLUTION MASS SPEC 7947 05:47:02,680 --> 05:47:03,920 INSTRUMENTATION. SO HERE ARE 7948 05:47:03,920 --> 05:47:07,680 BASED ON THIS IDEA WE CAN TRIPLE 7949 05:47:07,680 --> 05:47:10,360 THE THROUGH PUT OF ORIGINAL FOUR 7950 05:47:10,360 --> 05:47:12,520 COMPLEX SUGAR TAG TO GENERATE 12 7951 05:47:12,520 --> 05:47:16,400 TAG SUGAR TAG AND HERE SHOWS THE 7952 05:47:16,400 --> 05:47:19,640 RESOLVING POWER, 30,000 7953 05:47:19,640 --> 05:47:21,760 RESOLVING POWER WE CAN MICELY 7954 05:47:21,760 --> 05:47:23,360 RESOLVE THESE 12 QUANTIFICATION 7955 05:47:23,360 --> 05:47:27,840 CHANNEL. SO HERE SHOWS CENSUS, 7956 05:47:27,840 --> 05:47:32,560 WE CAN -- THIS WAS DONE WITH 7957 05:47:32,560 --> 05:47:33,360 GRAD SUNT IN THE LACK MIC LI. SO 7958 05:47:33,360 --> 05:47:37,320 WE CAN USE THE -- IN THE LAB 7959 05:47:37,320 --> 05:47:40,880 MIKE LI SO WE CAN GENERATE 12 7960 05:47:40,880 --> 05:47:44,960 COMPLEX SUGAR TAG. IF WE SWAP 7961 05:47:44,960 --> 05:47:48,080 CARBON 13 THERE IS A 6 7962 05:47:48,080 --> 05:47:48,960 MILIDALTON MASS DIFFERENCE 7963 05:47:48,960 --> 05:47:50,560 DIFFERENTIATED BUT IN LOW 7964 05:47:50,560 --> 05:47:51,880 RESOLUTION THIS MAY SEEM AS A 7965 05:47:51,880 --> 05:47:55,880 SINGLE PEAK, THE 115A ANDB 7966 05:47:55,880 --> 05:47:58,160 HOWEVER WAS HIGH RESOLUTION, WE 7967 05:47:58,160 --> 05:48:00,040 CAN RESOLVE THEM SO THIS IS 7968 05:48:00,040 --> 05:48:02,360 ACTUALLY SHOWING THE REQUIRED 7969 05:48:02,360 --> 05:48:03,760 MASS SPECTRAL RESOLUTION THAT 7970 05:48:03,760 --> 05:48:06,800 YOU CAN SYSTEMATICALLY RESOLVE 7971 05:48:06,800 --> 05:48:09,720 BASELINE RESOLVE THESE 12 7972 05:48:09,720 --> 05:48:13,200 REPORTER IONS YOU CAN SEE AGAIN 7973 05:48:13,200 --> 05:48:16,360 AT 30,000 OR EVEN 60,000 WE CAN 7974 05:48:16,360 --> 05:48:17,920 NICELY TELL THESE 12 CHANNELS 7975 05:48:17,920 --> 05:48:20,920 APART FROM EACH OTHER. HERE WE 7976 05:48:20,920 --> 05:48:24,360 ARE AGAIN TAKING FREELY RELEASED 7977 05:48:24,360 --> 05:48:28,920 BTG GLYCANS AND LOOK AT KNOWN 7978 05:48:28,920 --> 05:48:33,120 RATIO WITH THIS TALL PLEX SUGAR 7979 05:48:33,120 --> 05:48:35,560 TAG AND ON THE LOWER PANEL IS 7980 05:48:35,560 --> 05:48:38,080 EXAMPLE OF NUMBER OF GLYCANS IN 7981 05:48:38,080 --> 05:48:40,480 MSMS AND ALSO ZOOM IN FOR THE 7982 05:48:40,480 --> 05:48:42,360 QUANTIFICATION THAT ALLOWS US TO 7983 05:48:42,360 --> 05:48:44,160 QUANTIFY THESE THINGS. SO IN 7984 05:48:44,160 --> 05:48:46,080 THE REMAINING TIME I WANT TO 7985 05:48:46,080 --> 05:48:48,200 ALSO TELL YOU A LITTLE BIT ABOUT 7986 05:48:48,200 --> 05:48:49,920 BESIDES LOOKING AT FREELY 7987 05:48:49,920 --> 05:48:52,280 RELEASE GLYCANS WE CAN ALSO LOOK 7988 05:48:52,280 --> 05:48:54,800 AT INTACT GLYCOPEPTIDES IN 7989 05:48:54,800 --> 05:48:56,520 PARTICULAR THE GLYCOFORM BECAUSE 7990 05:48:56,520 --> 05:48:59,160 THAT'S ALSO IMPLICATED IN 7991 05:48:59,160 --> 05:49:03,040 DIFFERENT DISEASE CONDITIONS. 7992 05:49:03,040 --> 05:49:06,560 HERE WE TEAMED UP WITH WISCONSIN 7993 05:49:06,560 --> 05:49:07,560 ALZHEIMER'S DISEASE RESEARCH 7994 05:49:07,560 --> 05:49:09,560 CENTER AND WE DEVELOPED THIS 7995 05:49:09,560 --> 05:49:12,560 INTEGRATED ANALYTICAL WORK FLOW 7996 05:49:12,560 --> 05:49:14,640 TO LOOK AT GLYCOPROTEOME FROM 7997 05:49:14,640 --> 05:49:16,560 CEREBRAL SPINAL FLUID COLLECTED 7998 05:49:16,560 --> 05:49:18,360 AT DIFFERENT DISEASE STAGE AND 7999 05:49:18,360 --> 05:49:21,200 AFTER PROTEIN EXTRACTION 8000 05:49:21,200 --> 05:49:22,760 DIGESTION AND ENRICHMENT AND 8001 05:49:22,760 --> 05:49:25,000 ALSO WE HEARD ABOUT 8002 05:49:25,000 --> 05:49:26,560 FRAGMENTATION TECHNIQUE FOR 8003 05:49:26,560 --> 05:49:28,680 INTACT GLYCOPEPTIDE 8004 05:49:28,680 --> 05:49:31,320 QUANTIFICATION. HERE WE USING A 8005 05:49:31,320 --> 05:49:36,120 DIMANY METHYLATED LEUCINE TAG TO 8006 05:49:36,120 --> 05:49:37,480 QUANTIFY THE GLYCOPEP SIDES. 8007 05:49:37,480 --> 05:49:39,560 HERE IS GENERIC STRUCTURE IS A 8008 05:49:39,560 --> 05:49:43,760 MEAN REACTIVE GROUP WITH BALANCE 8009 05:49:43,760 --> 05:49:46,120 CARBONIL GROUP AND DIMETHYLATED 8010 05:49:46,120 --> 05:49:48,960 LEUCINE WITH STABLE ISOTOPES SO 8011 05:49:48,960 --> 05:49:51,880 HERE IS DUPLEX VERSION AND HERE 8012 05:49:51,880 --> 05:49:53,880 USING THE SAME HIGH RESOLUTION 8013 05:49:53,880 --> 05:49:55,680 MASS DEFECT PHENOMENON WE CAN 8014 05:49:55,680 --> 05:49:59,000 CREATE A 12 PLEX CHANNEL. THIS 8015 05:49:59,000 --> 05:50:03,960 12 PLEX WE CAN INTERROGATE 48 8016 05:50:03,960 --> 05:50:06,200 PATIENTS, DIVIDE INTO LEE 8017 05:50:06,200 --> 05:50:09,160 GROUPS, CONTROL MCI MILD 8018 05:50:09,160 --> 05:50:12,160 COGNITIVE IMPAIRMENT AND ADVANCE 8019 05:50:12,160 --> 05:50:13,720 DEMENTIA, 16 PATIENT EACH AND 8020 05:50:13,720 --> 05:50:18,600 WITH THIS INTEGRATED WORK FLOW 8021 05:50:18,600 --> 05:50:23,560 WE 12 MENTION QUANTIFY 8022 05:50:23,560 --> 05:50:26,040 GLYCOPEPTIDES TO 107 IN 8023 05:50:26,040 --> 05:50:26,440 GLYCOPROTEINS. 8024 05:50:26,440 --> 05:50:29,280 . WE LOOK AT THREE OF THESE 8025 05:50:29,280 --> 05:50:32,160 GLYCOPROTEINS RELATED TO AD. THE 8026 05:50:32,160 --> 05:50:34,160 GLYCANNER -- THE NEURAL CELL 8027 05:50:34,160 --> 05:50:36,080 ADHESION MOLECULE, SO THIS ONE 8028 05:50:36,080 --> 05:50:39,280 HAS FOUR DIFFERENT GLYCOSIDE AND 8029 05:50:39,280 --> 05:50:41,360 THERE IS MINIMUM DEGREE OF 8030 05:50:41,360 --> 05:50:43,360 GLYCAN HETEROGENEITY, COMPLIMENT 8031 05:50:43,360 --> 05:50:46,920 CS 3 IS NEURAL INFLAMMATORY 8032 05:50:46,920 --> 05:50:49,560 PROTEIN HAS SINGLE SITE ENCODE 8033 05:50:49,560 --> 05:50:52,960 22 DIFFERENT GLYCOFORMS. PERHAPS 8034 05:50:52,960 --> 05:50:53,960 MOST INTERESTING IS CLUSTERING 8035 05:50:53,960 --> 05:50:55,840 AND THIS ONE IS THE THIRD MOST 8036 05:50:55,840 --> 05:50:59,480 RISK GENE IN AD, AND ENCODES 449 8037 05:50:59,480 --> 05:51:02,040 AMINO ACID RESIDUE, FOUR 8038 05:51:02,040 --> 05:51:04,640 GLYCOSIDES AND SOME OF THESE 8039 05:51:04,640 --> 05:51:06,360 MINIMUM DEGREE OF GLYCAN 8040 05:51:06,360 --> 05:51:09,960 HETEROGENEITY BUT IMPORTANTLY 8041 05:51:09,960 --> 05:51:14,040 THIS 374 ENCODES 46 -- 56 8042 05:51:14,040 --> 05:51:15,760 GLYCOFORMS MANY HAVE SIALIC ACID 8043 05:51:15,760 --> 05:51:17,960 AND FUCOSYLATION. SO WE HAVE 8044 05:51:17,960 --> 05:51:21,560 ALSO LOOK AT GLOBALLY THESE 8045 05:51:21,560 --> 05:51:23,200 DISREGULATION OF M GLYCOPEPTIDES 8046 05:51:23,200 --> 05:51:26,640 IN THIS PANEL PLOT SHOWED AT THE 8047 05:51:26,640 --> 05:51:29,400 NCI AND NUMBER OF N GLYCOSIDES 8048 05:51:29,400 --> 05:51:32,560 AND GENE ONTOLOGY ANALYSIS 8049 05:51:32,560 --> 05:51:35,560 REVIEWED A GLYCOPROCESS 8050 05:51:35,560 --> 05:51:37,760 INCLUDING PLATELET DEGRANULATION 8051 05:51:37,760 --> 05:51:39,280 CELL ADHESION AND INFLAMMATORY 8052 05:51:39,280 --> 05:51:41,120 RESPONSE RELATED TO THE 8053 05:51:41,120 --> 05:51:43,520 MOLECULAR DEGENERATION. SO THEN 8054 05:51:43,520 --> 05:51:46,960 WE HAVE LOOK AT SOME OF THESE 8055 05:51:46,960 --> 05:51:50,240 GLYCOPEPTIDES AT NCI AND AD AND 8056 05:51:50,240 --> 05:51:52,960 THIS REALLY SHOWED NOT ONLY 8057 05:51:52,960 --> 05:51:55,440 REALLY THE SIGNIFICANT CHANGES 8058 05:51:55,440 --> 05:51:58,160 OCCURRING AT THE GLYCOSIDE, N 8059 05:51:58,160 --> 05:51:59,000 GLYCOPEPTIDES, NOT AT PROTEIN 8060 05:51:59,000 --> 05:52:01,440 LEVEL. SINCE I'M RUNNING SHORT 8061 05:52:01,440 --> 05:52:03,200 ON TIME I'M GOING TO CLOSE AND 8062 05:52:03,200 --> 05:52:08,000 TO POINT OUT THAT WE DIVIDE 8063 05:52:08,000 --> 05:52:09,000 DISEASE STAGE SPECIFIC IN 8064 05:52:09,000 --> 05:52:12,760 GLYCOPEPTIDES. SO TO WRAP UP I 8065 05:52:12,760 --> 05:52:15,800 HAVE SHOWN YOU BEDEVELOPED SET 8066 05:52:15,800 --> 05:52:17,520 OF GENERIC LABELING REAGENT THAT 8067 05:52:17,520 --> 05:52:19,080 ALLOWS US TO PERFORM HIGH 8068 05:52:19,080 --> 05:52:20,960 THROUGH PUT GLYCAN 8069 05:52:20,960 --> 05:52:22,640 QUANTIFICATION AND GLYCOPEPTIDE 8070 05:52:22,640 --> 05:52:25,600 QUANTIFYCATION. AND SO NUMBER 8071 05:52:25,600 --> 05:52:27,280 OF PUBLICATIONS ACKNOWLEDGING 8072 05:52:27,280 --> 05:52:30,160 THE UO 1 GRANT ALL TOGETHER, WE 8073 05:52:30,160 --> 05:52:32,560 IN THE PAST THREE YEARS 8074 05:52:32,560 --> 05:52:35,480 PUBLISHED 30 PUBLICATIONS AND 8075 05:52:35,480 --> 05:52:38,440 NUMBER OF THESE TALKS AND 8076 05:52:38,440 --> 05:52:42,040 SESSIONS IN PARTICULAR, THIS 8077 05:52:42,040 --> 05:52:43,920 UPCOMING JUNE WE HAVE THE NORTH 8078 05:52:43,920 --> 05:52:45,320 AMERICAN MASS SPEC SUMMER 8079 05:52:45,320 --> 05:52:47,560 SCHOOL, TALK MORE ABOUT OUR 8080 05:52:47,560 --> 05:52:50,400 TECHNOLOGY AND GREAT VARIETY OF 8081 05:52:50,400 --> 05:52:51,280 DIFFERENT COLLABORATIONINGS. 8082 05:52:51,280 --> 05:52:53,120 WITH THAT I WANT TO THANK YOU 8083 05:52:53,120 --> 05:52:57,600 ALL FOR KIND ATTENTION AND ALSO 8084 05:52:57,600 --> 05:52:59,280 AGAIN THANK YOU TO GLYCOCOMMON 8085 05:52:59,280 --> 05:53:00,480 FUNDS AND ALL OF YOU FOR YOUR 8086 05:53:00,480 --> 05:53:01,320 KIND ATTENTION. THANK YOU. 8087 05:53:01,320 --> 05:53:09,800 [APPLAUSE] 8088 05:53:09,800 --> 05:53:11,120 >> OUR NEXT SPEAKER IS 8089 05:53:11,120 --> 05:53:14,160 (OFF MIC) 8090 05:53:14,160 --> 05:53:18,440 LORI YANG. 8091 05:53:18,440 --> 05:53:35,760 (OFF MIC) 8092 05:53:35,760 --> 05:53:40,520 >> CAN EVERYONE HEAR ME? GREAT. 8093 05:53:40,520 --> 05:53:43,880 SO I WANT TO REITERATE OUR DEEP 8094 05:53:43,880 --> 05:53:45,720 GRATITUDE TOWARDS THE NIH AND 8095 05:53:45,720 --> 05:53:47,600 COMMON FUND OFFICIALS FOR ALL 8096 05:53:47,600 --> 05:53:49,280 THEIR SUPPORT THROUGHOUT THE 8097 05:53:49,280 --> 05:53:52,040 YEARS AND FOR MAKING THIS FINAL 8098 05:53:52,040 --> 05:53:54,920 MEETING HAPPEN AND IN PERSON. 8099 05:53:54,920 --> 05:53:57,320 FIRST LET ME TELL YOU A LITTLE 8100 05:53:57,320 --> 05:54:03,480 BIT ABOUT OUR COMPANY. WE ARE A 8101 05:54:03,480 --> 05:54:06,160 FOR PROFIT COMPANY. WE ARE AN 8102 05:54:06,160 --> 05:54:08,560 ACADEMIC SPIN OFF OF THE LAB OF 8103 05:54:08,560 --> 05:54:11,880 PROFESSOR ROB WOODS CO-FOUNDER 8104 05:54:11,880 --> 05:54:13,960 AND PRESIDENT OF LECTENZ BICO 8105 05:54:13,960 --> 05:54:15,520 YOU HEARD FROM EARLIER AND WILL 8106 05:54:15,520 --> 05:54:17,760 HEAR FROM AGAIN LATER. WE ARE 8107 05:54:17,760 --> 05:54:23,760 HEAD QUARTERED AT THE INNOVATION 8108 05:54:23,760 --> 05:54:25,400 GATEWAY INCUBATOR AT UNIVERSITY 8109 05:54:25,400 --> 05:54:26,880 OF GEORGIA. WE HAVE A SATELLITE 8110 05:54:26,880 --> 05:54:32,520 FACILITY IN SOUT SAN DIEGO 8111 05:54:32,520 --> 05:54:37,600 CALIFORNIA. SO WHAT IS 8112 05:54:37,600 --> 05:54:39,360 GLYCOSENSE? A SOLUBLE LECTIN 8113 05:54:39,360 --> 05:54:42,200 MICROARRAY BUT INSTEAD OF THE 8114 05:54:42,200 --> 05:54:44,280 LEPTONS BEING SPOTTED ON TO A 8115 05:54:44,280 --> 05:54:46,360 MICROSCOPE SLIDE, THEY ARE 8116 05:54:46,360 --> 05:54:48,520 CHEMICALLY CONJUGATED TO 8117 05:54:48,520 --> 05:54:53,200 MICROSPHERES AND SUSPENDED IN 8118 05:54:53,200 --> 05:54:56,560 SOLUTION. SO IT IS A MIX AND RUN 8119 05:54:56,560 --> 05:55:03,040 ASSAY.ASSAY. GLYCOSENSE BEADS ARE 8120 05:55:03,040 --> 05:55:04,480 MULTIPLEX MICROSPEARS WITH 8121 05:55:04,480 --> 05:55:06,560 DISCREET LEVELS OF RED 8122 05:55:06,560 --> 05:55:07,560 FLUORESCENCE INTENSITY AND EACH 8123 05:55:07,560 --> 05:55:09,760 IS CONJUGATED TO A DIFFERENT 8124 05:55:09,760 --> 05:55:13,800 GLYCAN BINDING PROTEIN. THE 8125 05:55:13,800 --> 05:55:17,000 BEADS ARE MIXED WITH FLUORESCENT 8126 05:55:17,000 --> 05:55:18,680 GLYCOPROTEIN ANOLYTE EITHER 8127 05:55:18,680 --> 05:55:22,760 DIRECTLY LABELED OR SUBSEQUENTLY 8128 05:55:22,760 --> 05:55:27,560 INCUBATED WITH SECONDARY 8129 05:55:27,560 --> 05:55:32,080 REAGENT. AS SHOWN HERE. AND THE 8130 05:55:32,080 --> 05:55:34,760 BEADS AND SAMPLE ARE THEN RUN ON 8131 05:55:34,760 --> 05:55:40,240 A FLOW SIGH TOM TEAR. HYDRO 8132 05:55:40,240 --> 05:55:41,800 DYNAMICALLY FOCUSES BEAD SUCH 8133 05:55:41,800 --> 05:55:44,560 THAT THEY RUN SINGLE FILE 8134 05:55:44,560 --> 05:55:47,400 THROUGH THIS ENLARGED FLOW CELL. 8135 05:55:47,400 --> 05:55:51,840 A LASER INTERROGATES EACH BEAD 8136 05:55:51,840 --> 05:55:53,080 BECAUSE IT IS JUST THE BEAD AND 8137 05:55:53,080 --> 05:55:56,360 NOT SURROUNDING SOLUTION, WASHES 8138 05:55:56,360 --> 05:55:59,040 ARE USUALLY UNNECESSARY. SO LET 8139 05:55:59,040 --> 05:56:01,360 ME TELL YOU A LITTLE BIT IN 8140 05:56:01,360 --> 05:56:04,320 DETAIL HOW WE GET FROM THE DATA 8141 05:56:04,320 --> 05:56:08,600 TO THE FINAL GLYCOPRINT. FIRST 8142 05:56:08,600 --> 05:56:11,680 LIGHT SCATTER IS MEASURED. FORD 8143 05:56:11,680 --> 05:56:13,440 AND SIDE SCANNER SORT OF RELATE 8144 05:56:13,440 --> 05:56:16,160 TO THE SIZE AND GRANULARITY OF A 8145 05:56:16,160 --> 05:56:17,680 CELL. BUT IN OUR CASE WE ARE 8146 05:56:17,680 --> 05:56:26,760 LOOKING AT BEADS. SO EACH DOT 8147 05:56:26,760 --> 05:56:28,560 REPRESENT AS BEAD. BECAUSE BEADS 8148 05:56:28,560 --> 05:56:32,120 ARE SO UNIFORM THEY CLUSTER 8149 05:56:32,120 --> 05:56:33,560 TIGHTLY AROUND THE CENTER. SO WE 8150 05:56:33,560 --> 05:56:39,280 DRAW A GATE TO ANALYZE SINGLE 8151 05:56:39,280 --> 05:56:40,920 BEAD AND NOT DUB BEADS OR 8152 05:56:40,920 --> 05:56:43,920 DEBRIS. THEN THE RED 8153 05:56:43,920 --> 05:56:46,960 FLUORESCENCE INTENSITY OF EACH 8154 05:56:46,960 --> 05:56:49,120 BEAD IS NEGATIVE. AS I SAID 8155 05:56:49,120 --> 05:56:52,520 BEFORE EACH BEAD SET HAS A 8156 05:56:52,520 --> 05:56:54,240 DEFINED AMOUNT OF RED DYE 8157 05:56:54,240 --> 05:56:55,760 INCORPORATED SHOWN SCHEMATICALLY 8158 05:56:55,760 --> 05:56:59,320 HERE AS A DIFFERENT LEVEL OF RED 8159 05:56:59,320 --> 05:57:05,640 COLOR. AND IT IS FIVE CP. SO BP 8160 05:57:05,640 --> 05:57:09,160 IS GATED, SO THAT THE GREEN 8161 05:57:09,160 --> 05:57:11,080 FLUORESCENCE OF EACH RED 8162 05:57:11,080 --> 05:57:11,760 FLUORESCENT BEAD CAN BE 8163 05:57:11,760 --> 05:57:16,360 MEASURED. AND WE SEE THAT ONLY 8164 05:57:16,360 --> 05:57:19,160 THE GAL AND GLCNAC DETECTING 8165 05:57:19,160 --> 05:57:22,760 BEADS HAVE A HIGH LEVEL OF GREEN 8166 05:57:22,760 --> 05:57:24,760 FLUORESCENCE IN THE X AXIS. AND 8167 05:57:24,760 --> 05:57:30,040 THAT'S BECAUSE THEY HAVE PROTEIN 8168 05:57:30,040 --> 05:57:31,520 BOUND ENLARGED ON THE RIGHT IT 8169 05:57:31,520 --> 05:57:33,000 HAS ONLY TERMINAL GAL AND 8170 05:57:33,000 --> 05:57:39,480 GLCNAC. TO SHOW ALL THE DATA ALL 8171 05:57:39,480 --> 05:57:43,160 TOGETHER HERE IS ANOTHER 8172 05:57:43,160 --> 05:57:47,120 BIVARIANT DOT PLOT, AGAIN, EACH 8173 05:57:47,120 --> 05:57:48,840 DOT REPRESENTS THE BEAD SO THE 8174 05:57:48,840 --> 05:57:50,760 BEADS ARE DISTINGUISHED BY THEIR 8175 05:57:50,760 --> 05:57:54,360 POSITION ALONG THE X AXIS WHILE 8176 05:57:54,360 --> 05:57:56,360 ANY GREEN FLUORESCENT 8177 05:57:56,360 --> 05:57:58,520 GLYCOPROTEIN BAND IS OBSERVED ON 8178 05:57:58,520 --> 05:58:04,640 THE Y AXIS AT THIS TIME AND ONLY 8179 05:58:04,640 --> 05:58:09,560 THE GAL AND GLCNAC HAVE 8180 05:58:09,560 --> 05:58:10,680 DETECTING BEADS AND NOT THE 8181 05:58:10,680 --> 05:58:12,160 OTHERS. SO THIS COB CONVERTED 8182 05:58:12,160 --> 05:58:15,240 INTO -- CONVERTED TO A SIMPLE 8183 05:58:15,240 --> 05:58:16,400 GLYCOPRINT, BAR GRAPH OF THE 8184 05:58:16,400 --> 05:58:18,960 GREEN FLUORESCENCE INTENSITY OF 8185 05:58:18,960 --> 05:58:22,040 EACH GLYCAN DETECTING BEAD. JUST 8186 05:58:22,040 --> 05:58:24,800 TO NOTE, THE RATIO IS 8187 05:58:24,800 --> 05:58:25,920 PURPOSEFULLY NOT ONE TO ONE 8188 05:58:25,920 --> 05:58:30,120 BECAUSE THIS IS A NOT AN 8189 05:58:30,120 --> 05:58:32,960 ABSOLUTELY QUANTITATIVE ASSAY, 8190 05:58:32,960 --> 05:58:36,360 EACH GLYCAN DETECTING REAGENT 8191 05:58:36,360 --> 05:58:40,600 HAS DIFFERENT SPECIFICITIES FOR 8192 05:58:40,600 --> 05:58:42,360 GLYCAN SO THIS ACTION IS BEST 8193 05:58:42,360 --> 05:58:45,040 FOR LOOKING AT CHANGES. ANOTHER 8194 05:58:45,040 --> 05:58:48,480 NOTE, IN REALITY THAT WAS A 8195 05:58:48,480 --> 05:58:52,360 SIMPLE UNIFORMLY GLYCOSYLATED 8196 05:58:52,360 --> 05:58:55,160 GLYCOPROTEIN. BUT IN ACTUALITY 8197 05:58:55,160 --> 05:58:56,560 GLYCOPROTEINS HAVE MULTIPLE 8198 05:58:56,560 --> 05:58:59,200 SITES OF GLYCOSYLATION AND 8199 05:58:59,200 --> 05:59:00,560 DIFFERENT GLYCOFORMS AT EACH 8200 05:59:00,560 --> 05:59:03,800 SITE. SO WHAT WE ARE LOOKING AT 8201 05:59:03,800 --> 05:59:05,960 IS A REPRESENTATION OF THE 8202 05:59:05,960 --> 05:59:08,400 MIXTURE OF GLYCOFORMS AND SENSE 8203 05:59:08,400 --> 05:59:10,600 N GLYCANS SHARE COMMON CORE WE 8204 05:59:10,600 --> 05:59:14,760 ARE ONLY INTERESTED MANY THE 8205 05:59:14,760 --> 05:59:20,960 CAPCAP. WE CHOSE FCSs PRESS TO 8206 05:59:20,960 --> 05:59:22,400 ANALYZE THE DATA BECAUSE THEY 8207 05:59:22,400 --> 05:59:23,560 OFFER A FREE VERSION FOR WHICH 8208 05:59:23,560 --> 05:59:25,560 WE CREATED THEM PLACE WITH 8209 05:59:25,560 --> 05:59:29,520 INTEGRATED SOPs SO THAT USERS 8210 05:59:29,520 --> 05:59:30,760 COULD UPLOAD THEIR DATA FILES 8211 05:59:30,760 --> 05:59:32,840 AND FOLLOW THE STEP BY STEP 8212 05:59:32,840 --> 05:59:36,320 INSTRUCTIONS FOR ANALYSIS BUT 8213 05:59:36,320 --> 05:59:37,760 USERS HAVEN'T REQUESTED OR 8214 05:59:37,760 --> 05:59:38,920 NEEDED THIS BECAUSE IT IS EASY 8215 05:59:38,920 --> 05:59:40,640 ENOUGH TO TAKE THE DATA OFF THE 8216 05:59:40,640 --> 05:59:45,720 INSTRUMENT. YOU CROSS VALIDATED 8217 05:59:45,720 --> 05:59:48,440 ASSAY ON OTHER SIGH TOM TEARS 8218 05:59:48,440 --> 05:59:52,280 USING CONTROLLED GLYCOPROTEINS 8219 05:59:52,280 --> 05:59:56,040 FLUORESCFLUORESCENT -- ALL THESE 8220 05:59:56,040 --> 05:59:58,600 INSTRUMENTS GAVE THE SAME DATA 8221 05:59:58,600 --> 06:00:01,240 EXCEPT THE NEXT WHICH IS AN 8222 06:00:01,240 --> 06:00:03,800 IMAGING CELL COUNTER. WE CROSS 8223 06:00:03,800 --> 06:00:04,960 COLLABORATED WITH COLLABORATORS 8224 06:00:04,960 --> 06:00:08,400 BY SENDING THEM KITS AND 8225 06:00:08,400 --> 06:00:09,160 FORTUNATELY THE CONTROL 8226 06:00:09,160 --> 06:00:11,560 EXPERIMENTS WORKED ON THEIR 8227 06:00:11,560 --> 06:00:15,080 INSTRUMENTS IN THEIR HANDS. AND 8228 06:00:15,080 --> 06:00:17,680 SO MAKE SURE WHAT REALLY CROSS 8229 06:00:17,680 --> 06:00:23,840 VALIDATED BY OTHER METHODS, TOP 8230 06:00:23,840 --> 06:00:26,560 ROW OR OUR GLYCOPRINTS. WE TOOK 8231 06:00:26,560 --> 06:00:30,320 FLUORESCENTLY LABELED FETERIN, A 8232 06:00:30,320 --> 06:00:35,080 HIGH LEVEL SIALIC ALPHA 26 AND 8233 06:00:35,080 --> 06:00:38,520 ALPHA 23 WITH SIALIDASE AND WE 8234 06:00:38,520 --> 06:00:41,560 SAW THAT THE SIALIDASE SIALIC 8235 06:00:41,560 --> 06:00:47,560 ACID LEVELS DECREASED WHILE THE 8236 06:00:47,560 --> 06:00:49,760 GALACTOSE LEVELS INCREASED AND 8237 06:00:49,760 --> 06:00:53,640 UPON ADDITION OF GAY LACTOSEISIT 8238 06:00:53,640 --> 06:00:56,320 THE GALACTOSE SIGNALS DECREASED 8239 06:00:56,320 --> 06:00:57,360 WHILE GLCNAC INCREASED FROM 8240 06:00:57,360 --> 06:01:00,760 BEING EXPOSED. CONSISTENT WITH 2 8241 06:01:00,760 --> 06:01:05,160 AB PROFILING IN MASS SPEC, THOSE 8242 06:01:05,160 --> 06:01:07,960 ARE THE GLYCOFORMS THAT ARE SEEN 8243 06:01:07,960 --> 06:01:11,840 IN ALL THREE POINTS. SO HERE 8244 06:01:11,840 --> 06:01:15,120 JUST A FEW APPLICATIONS OF 8245 06:01:15,120 --> 06:01:16,960 GLYCOSENSE YOU CAN DO SIMPLE 8246 06:01:16,960 --> 06:01:19,760 GLYCOPROTEIN ANALYSIS AND 8247 06:01:19,760 --> 06:01:21,280 GENERATE THESE GLYCOPRINTS, YOU 8248 06:01:21,280 --> 06:01:23,480 CAN MONITOR GLYCOPROTEIN 8249 06:01:23,480 --> 06:01:25,640 PRODUCTION IN PROCESS. AND YOU 8250 06:01:25,640 --> 06:01:29,120 CAN MONITOR GLYCOENGINEERING OF 8251 06:01:29,120 --> 06:01:31,160 GLYCOPROTEINS THAT IS 8252 06:01:31,160 --> 06:01:32,960 GLYCOSIDASE AND GLYCOSYL 8253 06:01:32,960 --> 06:01:34,960 TRANSFERASE REACTION. YOU CAN 8254 06:01:34,960 --> 06:01:37,080 ALSO LOOK AT THE GLYCOSYLATION 8255 06:01:37,080 --> 06:01:42,560 OF NATIVE ANTIBODIES. SO WE 8256 06:01:42,560 --> 06:01:45,520 HAVE USED FLUORESCENTLY LABELED 8257 06:01:45,520 --> 06:01:48,080 GLYCOPROTEINS FOR SIMPLICITY TO 8258 06:01:48,080 --> 06:01:51,600 HELP DEVELOP METHODS, ESPECIALLY 8259 06:01:51,600 --> 06:01:53,280 FETUIN BUT THIS WORKS WITH 8260 06:01:53,280 --> 06:01:55,200 INDIRECT DETECTION AS WELL. IF 8261 06:01:55,200 --> 06:02:00,320 WE TAKE UNLABELED FETUIN AND 8262 06:02:00,320 --> 06:02:04,520 INCUBATE WITH RB AND FOLLOWED BY 8263 06:02:04,520 --> 06:02:05,560 LABELED ANTI-FETUIN FAB THAT 8264 06:02:05,560 --> 06:02:07,560 FRAGMENT BECOMES ANTIBODIES 8265 06:02:07,560 --> 06:02:09,280 THEMSELVESES ARE GLYCOSYLATE WED 8266 06:02:09,280 --> 06:02:15,800 SEE THEY GIVE SIMILAR 8267 06:02:15,800 --> 06:02:17,680 GLYCOPRINTS. WE USE GLYCOMONITOR 8268 06:02:17,680 --> 06:02:19,360 GLYCOPRODUCTION PROTEIN IN 8269 06:02:19,360 --> 06:02:22,600 PROCESS, IN THIS CASE ROBO 1 8270 06:02:22,600 --> 06:02:24,840 GLYCOPROTEIN WHICH HAS A SINGLE 8271 06:02:24,840 --> 06:02:28,520 SITE N GLYCOSYLATION AND IS 8272 06:02:28,520 --> 06:02:30,840 CONVENIENTLY GFP TAGGED SO THAT 8273 06:02:30,840 --> 06:02:32,240 THE FLUORESCENT READ OUT IS 8274 06:02:32,240 --> 06:02:38,400 BUILT RIGHT IN. WE INCUBATED 8275 06:02:38,400 --> 06:02:39,760 GLYCOSENSE BEADS WITH CELL 8276 06:02:39,760 --> 06:02:42,560 CALLTURE MAINTENANCE, WE DIDN'T 8277 06:02:42,560 --> 06:02:44,520 PRE-PURIFY OR DO WASHINGS OR 8278 06:02:44,520 --> 06:02:45,880 RELEASE GLYCANS OR ANYTHING LIKE 8279 06:02:45,880 --> 06:02:48,720 THAT. ON THE LEFT THEY ARE ROBO 8280 06:02:48,720 --> 06:02:51,880 1 GLYCOPRINTS EACH DAY OVER 8281 06:02:51,880 --> 06:02:52,920 SEVEN DAYS OF PRODUCTION AND TO 8282 06:02:52,920 --> 06:02:58,960 LOOK AT IT ANOTHER WAY, ON THE 8283 06:02:58,960 --> 06:03:04,560 RIGHT IS A SIMPLE LINE GRAPH OF 8284 06:03:04,560 --> 06:03:05,600 EACH GLYCOFEATURE MONITORED OVER 8285 06:03:05,600 --> 06:03:08,920 IN A TIME COURSE. AND NORMALIZE 8286 06:03:08,920 --> 06:03:12,160 TO TOTAL PROTEIN AMOUNT. THANKS 8287 06:03:12,160 --> 06:03:18,600 TO KELLY MOREMEN FOR THE ROBO 1 8288 06:03:18,600 --> 06:03:21,600 AND PARASTOO AZADI FOR THE MASS 8289 06:03:21,600 --> 06:03:23,960 SPEC DATA. CONSISTENT WITH MASS 8290 06:03:23,960 --> 06:03:25,600 SPEC THERE IS A HIGH PERCENTAGE 8291 06:03:25,600 --> 06:03:32,400 OF TERMINALLY GLYCOSYLATED AND 8292 06:03:32,400 --> 06:03:34,360 FEWS TO LATED GLYCOPERFORMS AND 8293 06:03:34,360 --> 06:03:38,240 LOW PERCENTAGE OF U -- WE USE 8294 06:03:38,240 --> 06:03:40,440 GLYCOSENSE TO MONITOR 8295 06:03:40,440 --> 06:03:44,320 GLYCOENGINEERING. AND THIS IS 8296 06:03:44,320 --> 06:03:44,880 FETEUIN ANOTHER TIME COURSE 8297 06:03:44,880 --> 06:03:46,760 TREAT WITH SIALIDASE FOR ONE 8298 06:03:46,760 --> 06:03:49,040 HOUR FOLLOWED BY GALACTOSIGH 8299 06:03:49,040 --> 06:03:51,800 DAYS FOR TWO HOURS. YOU SEE THE 8300 06:03:51,800 --> 06:03:54,400 BEGINNING AND END POINTS OF THE 8301 06:03:54,400 --> 06:03:56,840 REACTION BUT HERE WE ARE 8302 06:03:56,840 --> 06:03:58,760 ACTUALLY FOLLOWING THE REACTION 8303 06:03:58,760 --> 06:04:02,840 AT 15 MINUTE INTERVALS SO YOU 8304 06:04:02,840 --> 06:04:06,040 CAN SEE THE SIALIC ACID SINGLE 8305 06:04:06,040 --> 06:04:07,320 PURPLE LINE DECREASE WITH 8306 06:04:07,320 --> 06:04:10,040 ADDITION OF SIALIDASE WHILE 8307 06:04:10,040 --> 06:04:11,960 GALACTOSE SIGNAL INCREASES. AND 8308 06:04:11,960 --> 06:04:15,840 THE YELLOW LINE AND UPON 8309 06:04:15,840 --> 06:04:18,720 ADDITION OF GALACTOSIDASE THE 8310 06:04:18,720 --> 06:04:23,560 GAL SIGNAL DECREASES WHILE 8311 06:04:23,560 --> 06:04:28,360 SIMULTANEOUSLY THE GLIK MA'AM 8312 06:04:28,360 --> 06:04:30,920 SIGNAL ON THE OPPOSITE LINE. SO 8313 06:04:30,920 --> 06:04:32,920 WE PUT GAL BACK ON OR SIALIC 8314 06:04:32,920 --> 06:04:34,920 ACID BACK ON SO ON THE LEFT SIDE 8315 06:04:34,920 --> 06:04:36,360 THE BEADS ARE SHOWN SIDE BY SIDE 8316 06:04:36,360 --> 06:04:38,640 TO SHOW THE DIFFERENCE BETWEEN 8317 06:04:38,640 --> 06:04:42,920 THE ENZYME AND UNTREATED AND 8318 06:04:42,920 --> 06:04:50,400 TREATED GLYCOPROTEIN. SO 8319 06:04:50,400 --> 06:04:51,080 AGALACTOASIALFET ORBSIN BLUE BAR 8320 06:04:51,080 --> 06:04:57,880 HAS HIGH LEVEL OF GLCNAC BUT 8321 06:04:57,880 --> 06:05:00,680 SIALYLTRANSFERASE THE GAL GOES 8322 06:05:00,680 --> 06:05:05,760 UP AND GOES DOWN TREATED BY GAL. 8323 06:05:05,760 --> 06:05:11,840 WE USED ALPHA 26 OR ALPHA 23 AND 8324 06:05:11,840 --> 06:05:15,360 WE SEE UPON ADDITION WITH THE 26 8325 06:05:15,360 --> 06:05:17,840 THAT THE 26 SIGNAL GOES UP, THE 8326 06:05:17,840 --> 06:05:20,560 ORANGE BAR, NOT THE 23 AND THE 8327 06:05:20,560 --> 06:05:24,840 GAL SIGNAL GOES DOWN. SIMILARLY 8328 06:05:24,840 --> 06:05:27,520 UPON TREATMENT WITH 23 8329 06:05:27,520 --> 06:05:28,840 SIALYLTRANSFERASE THE 2, 3 GRAY 8330 06:05:28,840 --> 06:05:31,400 BAR GOES UP, NOT 26 AND THE GAL 8331 06:05:31,400 --> 06:05:35,840 GOES DOWN. SO AGAIN NOT 8332 06:05:35,840 --> 06:05:38,040 QUANTITATIVE, NOT A ONE TO ONE 8333 06:05:38,040 --> 06:05:40,560 RELATIONSHIP. WE ALSO LOOKED AT 8334 06:05:40,560 --> 06:05:42,720 NATIVE ANTIBODY WHICH IS IS 8335 06:05:42,720 --> 06:05:43,520 DIFFICULT BECAUSE THEY DON'T 8336 06:05:43,520 --> 06:05:44,880 LOOK LIKE THIS, THEY LOOK LIKE 8337 06:05:44,880 --> 06:05:47,560 THIS. THE GLYCANS ARE SOMETIMES 8338 06:05:47,560 --> 06:05:48,640 INACCESSIBLE AND DIFFERENT 8339 06:05:48,640 --> 06:05:50,720 ANTIBODIES SUB CLASSES REACT 8340 06:05:50,720 --> 06:05:51,600 DIFFERENTLY IN OUR PHASE WE HAVE 8341 06:05:51,600 --> 06:05:55,840 TO TEST ON A CASE-BY-CASE BASIS. 8342 06:05:55,840 --> 06:05:57,200 SO IN THIS EXAMPLE HERE IS 8343 06:05:57,200 --> 06:05:59,960 PROTEIN G PURIFIED HUMAN IGG AND 8344 06:05:59,960 --> 06:06:01,400 TO MAKE SURE WHAT WE ARE SEEING 8345 06:06:01,400 --> 06:06:04,560 IS REAL, WE TOOK THE DIRECTLY 8346 06:06:04,560 --> 06:06:08,600 LABELED IGG IN BLUE, TREAT WITH 8347 06:06:08,600 --> 06:06:11,800 SIALIDASE, THE SIALIC ACID 8348 06:06:11,800 --> 06:06:14,360 SIGNAL GOES DOWN AND GAL SIGNAL 8349 06:06:14,360 --> 06:06:17,560 GOES UP AND UPON TREATMENT WITH 8350 06:06:17,560 --> 06:06:19,160 GALACTOSIGH DAYS THE GAL GOES 8351 06:06:19,160 --> 06:06:23,840 DOWN AND GLCNAC GOES BACK UP. 8352 06:06:23,840 --> 06:06:25,520 THANKS TO -- FOR THE SAMPLES AND 8353 06:06:25,520 --> 06:06:26,960 SHE RECEIVED AN ADMINISTRATIVE 8354 06:06:26,960 --> 06:06:28,240 SUPPLEMENT TO DO HIGH THROUGH 8355 06:06:28,240 --> 06:06:31,760 PUT ANALYSIS OF SIALYLATION OF 8356 06:06:31,760 --> 06:06:34,240 IGG FROM HUMAN AND MOUSE MODEL. 8357 06:06:34,240 --> 06:06:38,880 SO IN SUMMARY, IT IS AND IN 8358 06:06:38,880 --> 06:06:43,160 PROCESS GLYCO ANALYSIS METHOD 8359 06:06:43,160 --> 06:06:46,480 THAT CHARACTERIZES IT IS 8360 06:06:46,480 --> 06:06:48,600 MULTIPLEX RUN ALL THE BEADS AT 8361 06:06:48,600 --> 06:06:50,840 ONCE, MIX AND MATCH CHOOSE LEVEL 8362 06:06:50,840 --> 06:06:52,520 FEATURES YOU WANT TO DETECT, NO 8363 06:06:52,520 --> 06:06:54,280 NEED FOR RELEASE OF GLYCANS FROM 8364 06:06:54,280 --> 06:06:56,200 WASHING, VERY EASY TO USE. DON'T 8365 06:06:56,200 --> 06:06:58,560 NEED HIGH AMOUNT OF TRAINING, 8366 06:06:58,560 --> 06:07:00,200 SPECIAL EQUIPMENT OR SOFTWARE 8367 06:07:00,200 --> 06:07:05,440 JUST BASIC FLOW SIGH CYTOMETER. 8368 06:07:05,440 --> 06:07:08,480 AND FAST, CAN BE MONITORED IN 8369 06:07:08,480 --> 06:07:10,200 NEAR REAL TIME AFTER SHORT 8370 06:07:10,200 --> 06:07:11,880 BINDING STEP. WE DID LAUNCH THIS 8371 06:07:11,880 --> 06:07:15,400 PRODUCT IN IN JUNE OF 2020 AS 8372 06:07:15,400 --> 06:07:17,360 THE DIFFICULT TIME BUT IT 8373 06:07:17,360 --> 06:07:20,320 INCLUDES THE FOLLOWING BEADS, 8374 06:07:20,320 --> 06:07:22,880 BACKGROUND OF 23 GAL GLCNAC 8375 06:07:22,880 --> 06:07:24,880 DETECTING BEADS AND TUBE OF 8376 06:07:24,880 --> 06:07:26,880 GATING BEADS, THE FIVE 8377 06:07:26,880 --> 06:07:29,040 UNCONJUGATED BEADS. GLYCOPROTEIN 8378 06:07:29,040 --> 06:07:31,920 CONTROLS. AND THIS IS AVAILABLE 8379 06:07:31,920 --> 06:07:35,360 ON OUR WEB SHOP ALSO ON FISHER 8380 06:07:35,360 --> 06:07:36,280 SCIENTIFIC, YOU HAVE TO SEARCH 8381 06:07:36,280 --> 06:07:38,040 FOR THE PRODUCT AND FOR ANY UGA 8382 06:07:38,040 --> 06:07:39,640 PEOPLE LISTENING WE ARE ALL OUR 8383 06:07:39,640 --> 06:07:44,000 PRODUCTS ARE ON UGA MART AND FOR 8384 06:07:44,000 --> 06:07:46,240 INSTITUTIONS THAT USE THE JAGGER 8385 06:07:46,240 --> 06:07:46,920 PLATFORM ALL OUR PRODUCTS ARE ON 8386 06:07:46,920 --> 06:07:50,760 THERE. FINALLY I WOULD LIKE TO 8387 06:07:50,760 --> 06:07:53,520 THANK EVERYBODY PAST AND PRESENT 8388 06:07:53,520 --> 06:07:56,560 ON THE LECTENZ BIO TEAM, MATT 8389 06:07:56,560 --> 06:07:58,400 WHO DEVELOPED AND PERSONOR 8390 06:07:58,400 --> 06:07:59,920 COMMERCIALIZED THE KIT HE'S 8391 06:07:59,920 --> 06:08:03,880 MOVED ON BUT MELINA HAS 8392 06:08:03,880 --> 06:08:04,760 ADMIRALLY TAKEN UP PRODUCTION 8393 06:08:04,760 --> 06:08:07,400 AND OUR FUNDING SOURCES, 8394 06:08:07,400 --> 06:08:09,560 ESPECIALLY COMMON FUND TO 8395 06:08:09,560 --> 06:08:11,760 DEVELOP THIS PLATFORM AND 8396 06:08:11,760 --> 06:08:13,200 COMPANION SBIR TO DEVELOP THE 8397 06:08:13,200 --> 06:08:14,800 GLYCOENGINEERING APPLICATION. 8398 06:08:14,800 --> 06:08:16,880 ALL OF OUR BETA TESTERS 8399 06:08:16,880 --> 06:08:18,880 COLLABORATORS CUSTOMERS, CAN SEE 8400 06:08:18,880 --> 06:08:22,080 OUR PRODUCTS ON THE WEBSITE AND 8401 06:08:22,080 --> 06:08:23,360 FOR COLLABORATIONS OR QUESTIONS 8402 06:08:23,360 --> 06:08:25,160 PLEASE CONTACT ME DIRECTLY OR 8403 06:08:25,160 --> 06:08:27,760 VISIT US AT OUR POSTERS OUTSIDE. 8404 06:08:27,760 --> 06:08:28,720 THANKS VERY MUCH FOR YOUR 8405 06:08:28,720 --> 06:08:29,360 ATTENTION. 8406 06:08:29,360 --> 06:08:54,920 [APPLAUSE] 8407 06:08:54,920 --> 06:08:59,640 THANK YOU, PAM FOR LEADING SO 8408 06:08:59,640 --> 06:09:01,080 WELL AND ORGANIZING THIS MEETING 8409 06:09:01,080 --> 06:09:04,480 AND BY THE WAY HOW WAS DINNER 8410 06:09:04,480 --> 06:09:10,280 LAST NIGHT? WHAT ARE LECTENZ, 8411 06:09:10,280 --> 06:09:12,280 NORMAL CLASS OF TARGETING 8412 06:09:12,280 --> 06:09:14,160 REAGENTS THAT ARE LECTIN LIKE 8413 06:09:14,160 --> 06:09:16,120 AND ENZYME DERIVED. MADE FROM 8414 06:09:16,120 --> 06:09:17,000 ENZYME BECAUSE THEY ARE MADE 8415 06:09:17,000 --> 06:09:20,880 FROM ENZYME THEY ARE DIFFERENT 8416 06:09:20,880 --> 06:09:24,160 FROM NATURAL LECTENZ IN MANY 8417 06:09:24,160 --> 06:09:25,480 WAYS AND THEY HAVE TUNEABLE 8418 06:09:25,480 --> 06:09:28,240 AFFINITIES IN BINDING KINETICS. 8419 06:09:28,240 --> 06:09:31,840 AND THOSE ARE ESSENTIAL FOR 8420 06:09:31,840 --> 06:09:36,800 AFFINITY OF PUBLICATION, ON THE 8421 06:09:36,800 --> 06:09:38,720 ENZYME PROPERTY AND THEY ARE 8422 06:09:38,720 --> 06:09:41,160 MONOMERIC AND THEY ARE 8423 06:09:41,160 --> 06:09:45,200 MONOMERIC, MINIMIZE THE -- 8424 06:09:45,200 --> 06:09:46,880 THROUGH CELLTURE CELLS, THAT'S 8425 06:09:46,880 --> 06:09:51,080 GOOD FOR STAINING OF LIVE CELLS. 8426 06:09:51,080 --> 06:09:52,320 THEY ARE COMMONLY EXPRESSED IN 8427 06:09:52,320 --> 06:09:56,560 E. COLI SO PRODUCE LARGE AMOUNT 8428 06:09:56,560 --> 06:10:02,680 AND MINIMIZING -- DURATION AND 8429 06:10:02,680 --> 06:10:05,160 MOST IMPORTANTLY IN THEORY, 8430 06:10:05,160 --> 06:10:07,720 EVERY SINGLE CARBOHYDRATE 8431 06:10:07,720 --> 06:10:09,360 PROCESS CAN BE CONVERTED INTO 8432 06:10:09,360 --> 06:10:14,760 LECTENZ. HOW THEY ARE MADE. 8433 06:10:14,760 --> 06:10:16,480 SIMPLY THEY ARE COMPUTATIONAL 8434 06:10:16,480 --> 06:10:18,640 GUIDED DIRECTLY EVOLUTION. THEY 8435 06:10:18,640 --> 06:10:20,760 ARE MADE AND START WITH CAUSE OF 8436 06:10:20,760 --> 06:10:22,200 ENZYME WITH KNOWN STRUCTURE, WE 8437 06:10:22,200 --> 06:10:25,440 ALL KNOW THAT, AND USE THE 8438 06:10:25,440 --> 06:10:27,040 COMPUTATIONAL MODEL TO IDENTIFY 8439 06:10:27,040 --> 06:10:29,760 SITES TO MUTATE AND WE USE THOSE 8440 06:10:29,760 --> 06:10:35,800 DATA MODERN DATA TO DESIGN 8441 06:10:35,800 --> 06:10:37,760 COMMON TORRIAL LIBRARY AND 8442 06:10:37,760 --> 06:10:42,600 EXPRESS -- COMBINATORIAL LIBRARY 8443 06:10:42,600 --> 06:10:49,920 AND OTHER RUNGS OF MY OPINION 8444 06:10:49,920 --> 06:10:53,160 AND WE IDENTIFY THOSE HIGH 8445 06:10:53,160 --> 06:10:55,440 BINDERS, EXPRESS, WE TAKE THOSE 8446 06:10:55,440 --> 06:10:58,480 HIGH BINDERS, GROUND THEM IN E. 8447 06:10:58,480 --> 06:11:00,640 COLI AND HAVE THE PROTEIN 8448 06:11:00,640 --> 06:11:05,440 VALIDATED FOR BINDING AFFINITY 8449 06:11:05,440 --> 06:11:06,480 AND BINDING SPECIFICITY. OF 8450 06:11:06,480 --> 06:11:10,560 COURSE WE WANT TO PUT THOSE 8451 06:11:10,560 --> 06:11:11,560 TOIUS BY BETA TESTING 8452 06:11:11,560 --> 06:11:15,640 APPLICATIONS. MODERATE IS KEY 8453 06:11:15,640 --> 06:11:16,960 TO LECTENZ DEVELOPMENT. THIS IS 8454 06:11:16,960 --> 06:11:24,000 JUST ONE OF EXAMPLE, COMPLEX 8455 06:11:24,000 --> 06:11:25,520 CRYSTAL STRUCTURE WITH LIGAND 8456 06:11:25,520 --> 06:11:30,480 AND IF NOT THERE IS -- SOME OF 8457 06:11:30,480 --> 06:11:35,960 THEM WILL BE RUN THREE OR FOUR 8458 06:11:35,960 --> 06:11:38,960 WEEKSWEEKS. COME UP WITH THREE GROUPS 8459 06:11:38,960 --> 06:11:43,760 OF RESIDUES AND RED ONE IS THE 8460 06:11:43,760 --> 06:11:46,240 ONE THAT FAVORABLE RESIDUE THOSE 8461 06:11:46,240 --> 06:11:48,160 ARE INVOLVED IN ENZYME ACTIVITY 8462 06:11:48,160 --> 06:11:49,600 WE DON'T NEED SO WE WANT TO 8463 06:11:49,600 --> 06:11:51,560 MUTATE THOSE. AND ANOTHER GROUP 8464 06:11:51,560 --> 06:11:54,160 IS THE BLUE ONE WE IDENTIFY, 8465 06:11:54,160 --> 06:11:58,560 THOSE ARE CALLED TEPID RESIDUES. 8466 06:11:58,560 --> 06:12:02,480 SO CLOSE TO LIGAND BUT UNDER 8467 06:12:02,480 --> 06:12:05,600 5-ANGSTROM OR LESS. BUT THEY 8468 06:12:05,600 --> 06:12:06,960 BARELY CONTRIBUTE ENERGY TO THE 8469 06:12:06,960 --> 06:12:10,280 BINDING. SO THOSE ARE THE ONE WE 8470 06:12:10,280 --> 06:12:13,720 MUTATE ONE BY ONE EACH O OTHER 8471 06:12:13,720 --> 06:12:14,760 ASSAYS AND EVERYTHING COMBINED 8472 06:12:14,760 --> 06:12:20,040 TO MAKE LIBRARY. THIS IS JUST 8473 06:12:20,040 --> 06:12:24,160 ONE EXAMPLE OF WE START WITH 8474 06:12:24,160 --> 06:12:28,280 ENGINEER SIALIC ACID LECTINS BY 8475 06:12:28,280 --> 06:12:31,200 ENDURING FROM METHYL SAL DAYS B, 8476 06:12:31,200 --> 06:12:32,480 CAME ONE RUNGS AFTER GOING 8477 06:12:32,480 --> 06:12:34,520 THROUGH PROCESS AND WE CAME UP 8478 06:12:34,520 --> 06:12:39,000 WITH TWO LECTINS ONE WE CALL 8479 06:12:39,000 --> 06:12:43,640 TWO, THREE SPECIFIC, OTHER IS 8480 06:12:43,640 --> 06:12:46,920 PAIN SPECIFICKER 5 GC AND 8481 06:12:46,920 --> 06:12:48,320 LINKAGE INDEPENDENT MANNER, IT 8482 06:12:48,320 --> 06:12:53,040 WILL BIND TO OTHER 2, 3, HOUR 23 8483 06:12:53,040 --> 06:12:57,520 AND HOUR 28. THIS SHOWS HOW WE 8484 06:12:57,520 --> 06:12:59,800 MADE THE BINDING CONSTANT, THIS 8485 06:12:59,800 --> 06:13:02,160 IS BY MEASUREMENT AND EVERYTHING 8486 06:13:02,160 --> 06:13:05,480 IS ONE MICROMOLAR OR SMALLER, 8487 06:13:05,480 --> 06:13:09,720 BINDING CONSTANT AND TARGET OF 8488 06:13:09,720 --> 06:13:17,600 LEPTONS. THIS IS WE WANT TO SEE 8489 06:13:17,600 --> 06:13:20,600 HOW SPECIFIC REAGENTS ARE, ONE 8490 06:13:20,600 --> 06:13:30,200 BINDING ARRAY, IN THE COLUMN IN 8491 06:13:30,200 --> 06:13:31,360 THE LAB IN UNIVERSITY AND GOING 8492 06:13:31,360 --> 06:13:34,440 TO SHOW THE DATA FROM HER LAB 8493 06:13:34,440 --> 06:13:40,080 BUT WE ALSO HAD GET SURVEYS FROM 8494 06:13:40,080 --> 06:13:42,080 NCFG AND Z BIOTECH AND THEY 8495 06:13:42,080 --> 06:13:43,280 PRODUCE HAVE SIMILAR RESULTS. 8496 06:13:43,280 --> 06:13:46,600 WE CAN SEE THAT, SPECIFICALLY 8497 06:13:46,600 --> 06:13:50,600 BIND TO OTHER 23, 26, 28 AND 8498 06:13:50,600 --> 06:13:53,920 IDENTIFY A 5 AC SO TWO DAY PAN 8499 06:13:53,920 --> 06:13:56,480 SPECIFIC. ONE THIS RUNS ONE DOWN 8500 06:13:56,480 --> 06:13:58,600 SIDE OF THIS REAGENT IS THAT IT 8501 06:13:58,600 --> 06:14:03,960 IS NOT REALLY (INAUDIBLE) 8502 06:14:03,960 --> 06:14:05,440 SENSITIVE O -- WE HAD TO DO 8503 06:14:05,440 --> 06:14:07,960 BINDING DETECTION IN THE VERY 8504 06:14:07,960 --> 06:14:09,080 LOW SAM, INCREASE SALT SIGNAL 8505 06:14:09,080 --> 06:14:15,160 WOULD BE REDUCED. SO WE WENT TO 8506 06:14:15,160 --> 06:14:17,120 DO ANOTHER RUNS OF MUTAGENESIS 8507 06:14:17,120 --> 06:14:19,160 AND THIS COME UP WITH VERSION 2, 8508 06:14:19,160 --> 06:14:21,880 SO THIS IS VERSION 2 OF THESE 8509 06:14:21,880 --> 06:14:25,400 PAN SPECIFIC NOW SIGNAL HAS BEEN 8510 06:14:25,400 --> 06:14:27,280 INCREASED ABOUT 200 TIMES. LESS 8511 06:14:27,280 --> 06:14:30,760 SENSITIVITY. SO IT IS ON THE -- 8512 06:14:30,760 --> 06:14:32,160 THESE SHOW YOU THE SPECIFICITY 8513 06:14:32,160 --> 06:14:40,360 IS THE SAME, STILL SPECIFIC. 23 8514 06:14:40,360 --> 06:14:42,920 SPECIFIC. HOWEVER WHEN THESE 8515 06:14:42,920 --> 06:14:44,480 LEVELS AMONG OTHER THINGS. 8516 06:14:44,480 --> 06:14:48,080 COUPLE OF EXAMPLES WITH WESTERN 8517 06:14:48,080 --> 06:14:50,360 BLOT, VERY SPECIFIC 23, HOUR 23 8518 06:14:50,360 --> 06:14:53,960 AND PAN SPECIFIC AND THESE ALPHA 8519 06:14:53,960 --> 06:14:58,560 26 IS NOT LECTENZ, IT IS 8520 06:14:58,560 --> 06:15:03,680 ENGINEERED FROM MUCH ROOM 8521 06:15:03,680 --> 06:15:08,200 LEPTON. COMPLETE SPECIFICITY. 8522 06:15:08,200 --> 06:15:12,960 THIS IS BRIGHT ONE IHC AND DATA 8523 06:15:12,960 --> 06:15:14,640 ALPHA 23 SPECIFIC BINDS MORE 8524 06:15:14,640 --> 06:15:18,280 QUITE SEEMS TO FOCUS ON THESE 8525 06:15:18,280 --> 06:15:20,800 VARIOUS CELLS AND THESE OTHER 8526 06:15:20,800 --> 06:15:25,360 TWO IS CONFINED MOSTLY TO THIS 8527 06:15:25,360 --> 06:15:30,880 LAMINA PROPRIA. AND IF WE TREAT 8528 06:15:30,880 --> 06:15:35,560 WITH NEUROLAMIDASE THE SIGNAL IS 8529 06:15:35,560 --> 06:15:37,080 GONE FORMAL 23 AND ALPHA 26 IF 8530 06:15:37,080 --> 06:15:39,800 YOU STAIN WITH ECA AFTER ALPHA 8531 06:15:39,800 --> 06:15:41,440 23 SIGNAL INCREASE, BECAUSE YOU 8532 06:15:41,440 --> 06:15:44,280 GET RID OF SIALIC ACID AND THE 8533 06:15:44,280 --> 06:15:46,720 -- GET EXPOSED SO YOU SEE MUCH 8534 06:15:46,720 --> 06:15:50,120 MOVE SIGNAL ESPECIALLY IN THE 8535 06:15:50,120 --> 06:15:54,200 AREAS YOU SEE SIGNAL ECA, OVER 8536 06:15:54,200 --> 06:16:01,280 THE UNTREATED. SIGNAL THERE. 8537 06:16:01,280 --> 06:16:08,760 THIS IS RESULT FROM COMMON 8538 06:16:08,760 --> 06:16:10,680 CENTER. AND ON THIS PICTURE YOU 8539 06:16:10,680 --> 06:16:21,840 -- THOSE ARE SKIN CELLS AND 8540 06:16:21,840 --> 06:16:22,760 SIALYLTRANSFERASE, IT IS GONE. 8541 06:16:22,760 --> 06:16:26,840 SO IN THE CASE OF STOICHIOMETRY 8542 06:16:26,840 --> 06:16:28,960 IT IS CLEAR WE TREAT WITH 8543 06:16:28,960 --> 06:16:31,160 NEURAMINIDASE, A SIGNAL IS GONE. 8544 06:16:31,160 --> 06:16:33,560 IT BECOMES RED ONE MEANS THERE 8545 06:16:33,560 --> 06:16:35,880 IS NO BINDING. THIS IS ON THE 8546 06:16:35,880 --> 06:16:39,280 LIVE CELLS CULTURE CELLS. OKAY. 8547 06:16:39,280 --> 06:16:43,040 THE APPLICATION CAN ALSO USE 8548 06:16:43,040 --> 06:16:45,360 AFFINITY PURIFICATION AND THIS 8549 06:16:45,360 --> 06:16:48,480 IS ONE WE MIX EQUAL AMOUNT OF 8550 06:16:48,480 --> 06:16:52,440 HOUR 23 AND ALPHA 26 8551 06:16:52,440 --> 06:16:53,560 GLYCOPROTEINS AND THE WESTERN 8552 06:16:53,560 --> 06:16:56,120 BLOT SHOW YOU THERE IS A CLEAR 8553 06:16:56,120 --> 06:17:02,880 SEPARATION HERE OF THESE TWO. SO 8554 06:17:02,880 --> 06:17:06,400 ALPHA 23, THIS WILL BE ALPHA 2, 8555 06:17:06,400 --> 06:17:08,760 6, THIS IS ALPHA 2, 3 AND 8556 06:17:08,760 --> 06:17:10,080 WESTERN BLOT SHOWS RESULT IN 8557 06:17:10,080 --> 06:17:19,680 HERE. WE ALSO USE OUR REAGENTS. 8558 06:17:19,680 --> 06:17:21,960 THIS IS ANOTHER EXAMPLE WE USE 8559 06:17:21,960 --> 06:17:28,400 NOVEL ONE EXPRESS HK 293 CELLS 8560 06:17:28,400 --> 06:17:29,920 PROVIDED BY (INAUDIBLE) YOU CAN 8561 06:17:29,920 --> 06:17:32,560 SEE THAT WE CAN HAVE THE PROTEIN 8562 06:17:32,560 --> 06:17:35,280 PURIFIED, COME OUT WITH ABOUT 8563 06:17:35,280 --> 06:17:41,120 20% OF TOTAL AMOUNT OF EXPRESS 8564 06:17:41,120 --> 06:17:43,840 AND WESTERN BLOT SHOWS THAT IN 8565 06:17:43,840 --> 06:17:47,560 THE FLOW THROUGH IN THE FLOW 8566 06:17:47,560 --> 06:17:50,040 THROUGH YOU SEE ILLUSION WITH 8567 06:17:50,040 --> 06:17:52,240 ALPHA 2, 3 SPECIFIC PROBE IS 8568 06:17:52,240 --> 06:17:55,240 USED IT IS USED YOU SEE MOST OF 8569 06:17:55,240 --> 06:17:57,840 THEM SIGNAL COMES BECOMES AN 8570 06:17:57,840 --> 06:18:02,360 ILLUSION A LITTLE BIT MANY THE IN THE 8571 06:18:02,360 --> 06:18:06,160 INPUT IF YOU USE ALPHA 26 PROBE 8572 06:18:06,160 --> 06:18:09,040 YOU SEE FLOW THREE MOSTLY IS 8573 06:18:09,040 --> 06:18:12,120 ALPHA 2, 6 BUT ALSO DETECT ALPHA 8574 06:18:12,120 --> 06:18:16,160 2 ON THE ILLUSION AS WELL THIS 8575 06:18:16,160 --> 06:18:19,120 IS BECAUSE FRACTION CONTAINS 8576 06:18:19,120 --> 06:18:21,160 THOSE ALPHA 2, 3 AND ALPHA 2, 6 8577 06:18:21,160 --> 06:18:26,360 IN SINGLE. (INAUDIBLE) LAB DID 8578 06:18:26,360 --> 06:18:27,760 SERVICE FOR US AND YOU CAN SEE 8579 06:18:27,760 --> 06:18:29,760 THAT THERE IS A CLEAR ENRICHMENT 8580 06:18:29,760 --> 06:18:33,960 OF SIALYLATION ENRICHMENT. 8581 06:18:33,960 --> 06:18:35,640 BECAUSE WE HAD TO MAKE ENZYME 8582 06:18:35,640 --> 06:18:39,400 FROM THERE, WE HAD TO START WITH 8583 06:18:39,400 --> 06:18:44,560 ENZYME TO MAKE THE LECTENZ SO 8584 06:18:44,560 --> 06:18:45,760 EXPRESS IN WAY HIGH AUTHORITY 8585 06:18:45,760 --> 06:18:48,000 WITH LOW COST SO WE DECIDED TO 8586 06:18:48,000 --> 06:18:49,360 MAKE THOSE LEVELS OF 8587 06:18:49,360 --> 06:18:50,720 GLYCOSCIENCE COMMUNITY AND THAT 8588 06:18:50,720 --> 06:18:54,760 HE ARE ON OUR WEBSITE. THANK 8589 06:18:54,760 --> 06:18:58,800 YOU TO THE GROUP. THIS IS OUR 8590 06:18:58,800 --> 06:19:00,720 GROUP, I WANT TO THANK THE PAST 8591 06:19:00,720 --> 06:19:02,160 MEMBERS AND OUR COLLABORATOR AND 8592 06:19:02,160 --> 06:19:03,800 SERVICE PROVIDERS. THANK YOU 8593 06:19:03,800 --> 06:19:04,240 VERY MUCH. 8594 06:19:04,240 --> 06:19:12,520 [APPLAUSE] 8595 06:19:12,520 --> 06:19:13,480 >>STUDENT: SOFT (INAUDIBLE) THE 8596 06:19:13,480 --> 06:19:35,640 LAST TALK. 8597 06:19:35,640 --> 06:19:38,560 >> 3-D STRUCTURE. WHAT YOU SEE 8598 06:19:38,560 --> 06:19:41,560 IS AN MD SIMULATION OF CAPSID 8599 06:19:41,560 --> 06:19:43,640 POLYSACCHARIDE FRAGMENT AND WHAT 8600 06:19:43,640 --> 06:19:45,480 IT REALLY POINTS OUT AL HOE WE 8601 06:19:45,480 --> 06:19:48,160 AS GLYCOBIOLOGISTS SAY 8602 06:19:48,160 --> 06:19:48,760 CARBOHYDRATE ARE FLEXIBLE IT IS 8603 06:19:48,760 --> 06:19:50,560 NOT ALWAYS CLEAR THAT WE ARE 8604 06:19:50,560 --> 06:19:52,360 TRULY HAVE THAT IN OUR HEADS. 8605 06:19:52,360 --> 06:19:53,840 BECAUSE WE WILL SAY AND I GET 8606 06:19:53,840 --> 06:19:55,320 ASKED THIS ALL THE TIME, WHAT IS 8607 06:19:55,320 --> 06:19:58,280 THE SHAPE OF MY SUGAR? AND I 8608 06:19:58,280 --> 06:20:00,360 WILL SAY IT IS FLEXIBLE, I KNOW 8609 06:20:00,360 --> 06:20:03,400 WHAT IS ITS SHAPE? SO YOU HAVE 8610 06:20:03,400 --> 06:20:08,560 TO THINK OF SHAPE AS AN ENSEMBLE 8611 06:20:08,560 --> 06:20:09,480 OF PROBABLE SHAPES. THE MOST 8612 06:20:09,480 --> 06:20:10,720 COMMON SHAPE AND OTHER SHAPES. 8613 06:20:10,720 --> 06:20:15,280 ONE OF THE CHALLENGES WE FACE IS 8614 06:20:15,280 --> 06:20:16,160 HOW DO YOU REPRESENT IN A VIDEO 8615 06:20:16,160 --> 06:20:17,520 IT IS GREAT BUT YOU DON'T 8616 06:20:17,520 --> 06:20:20,160 PUBLISH A VIDEO. YOU NEED SOME 8617 06:20:20,160 --> 06:20:22,800 OTHER WAY. WHAT YOU SEE RIGHT 8618 06:20:22,800 --> 06:20:25,480 SIDE IS A MOSS PLAT PLOT BECAUSE 8619 06:20:25,480 --> 06:20:27,120 SUGARS ON SURFACE LOOK LIKE 8620 06:20:27,120 --> 06:20:28,880 MOSS, THIS IS AN OVERLAY OF 8621 06:20:28,880 --> 06:20:31,320 MULTIPLE SHAPES FROM AN MD 8622 06:20:31,320 --> 06:20:32,960 SIMULATION. SO JUST A WAY TO 8623 06:20:32,960 --> 06:20:34,800 REPRESENT THINGS BUT THIS IS A 8624 06:20:34,800 --> 06:20:36,200 REALM OF SPECIALIST AND OUR 8625 06:20:36,200 --> 06:20:40,160 WHOLE GOAL OF THIS TOOL WE HAVE 8626 06:20:40,160 --> 06:20:41,160 DEVELOPED AND ARE CONTINUING TO 8627 06:20:41,160 --> 06:20:42,600 DEVELOP IS TO GET IN THE HANDS 8628 06:20:42,600 --> 06:20:48,200 OF THE NON-SPECIALISTS. SO HERE 8629 06:20:48,200 --> 06:20:50,720 IS OUR WEBSITE I POINTED OUT 8630 06:20:50,720 --> 06:20:53,160 EARLIER PDB SEARCH, WHAT I WILL 8631 06:20:53,160 --> 06:20:54,840 TALK TO YOU ABOUT NOW IS HOW DO 8632 06:20:54,840 --> 06:21:01,760 YOU MODEL THE SHAPES OF SUGAR. 8633 06:21:01,760 --> 06:21:03,480 THREE WAYS TO DO THIS ONE IS THE 8634 06:21:03,480 --> 06:21:06,000 POINT AND CLICK THAT WE SHOWED 8635 06:21:06,000 --> 06:21:08,160 YOU GRAPHICAL USER ENTERPACE AND 8636 06:21:08,160 --> 06:21:10,120 THAT IS THE WAY I MOST COMMONLY 8637 06:21:10,120 --> 06:21:11,560 DO IT BUT THERE ARE OTHERS AND 8638 06:21:11,560 --> 06:21:15,360 ONE WAS EARLY ADOPTED BY CFG 8639 06:21:15,360 --> 06:21:17,240 WHICH IS PASSING THROUGH THE URL 8640 06:21:17,240 --> 06:21:20,000 STREAM SO WHEN YOU RUN HTTP BLAH 8641 06:21:20,000 --> 06:21:21,920 BLAH BLAH SLASH DA, DA, DA. 8642 06:21:21,920 --> 06:21:23,680 THERE IS A WAY THE DO THAT AND 8643 06:21:23,680 --> 06:21:25,320 YOU CAN ENCODE IT AND IT WILL 8644 06:21:25,320 --> 06:21:26,960 SENSE SEQUENCE OF SUGAR TO OUR 8645 06:21:26,960 --> 06:21:30,320 SITE AND BUILD IT. THERE IS A 8646 06:21:30,320 --> 06:21:33,960 MORE SOPHISTICATED WAY TO ENCODE 8647 06:21:33,960 --> 06:21:36,160 MORE INFORMATION THAN API. THIS 8648 06:21:36,160 --> 06:21:41,680 IS ADOPTED BY GLYCAN THAT ALLOWS 8649 06:21:41,680 --> 06:21:44,320 REMOTE WEB SITES OR DATABASES TO 8650 06:21:44,320 --> 06:21:46,880 GENERATE 3-D STRUCTURE 8651 06:21:46,880 --> 06:21:51,160 AUTONOMOUSLY. IN TERMS OF 8652 06:21:51,160 --> 06:21:53,480 USAGE, THE TOOLS HAVE BEEN VERY 8653 06:21:53,480 --> 06:21:58,280 WIDELY USED. IF WE LOOK BACK SAY 8654 06:21:58,280 --> 06:22:02,480 2014 WE HAD MAYBE 20 PAPERS 8655 06:22:02,480 --> 06:22:07,360 SITING EITHER TERM GLYCAN WEB OR 8656 06:22:07,360 --> 06:22:09,760 TERM GLYCAN.ORG. NOW WE ARE ON 8657 06:22:09,760 --> 06:22:12,000 PROJECTED TO HIT ABOUT 100 8658 06:22:12,000 --> 06:22:14,880 PAPERS. OF THAT COUPLE ARE MINE. 8659 06:22:14,880 --> 06:22:17,240 I HAVEN'T FILTERED THIS OUT BUT 8660 06:22:17,240 --> 06:22:20,960 CERTAINLY NOT THAT MANY. YOU 8661 06:22:20,960 --> 06:22:26,080 HAVE SEEN THAT ABOUT THE SAME 8662 06:22:26,080 --> 06:22:27,600 AMOUNT MENTION GLEE CAM WEB OR 8663 06:22:27,600 --> 06:22:30,680 GLEE CAM.ORG. THE ONES SAYING 8664 06:22:30,680 --> 06:22:33,240 WEB ALSO AGREE CAM.ORG. BUT THEY 8665 06:22:33,240 --> 06:22:35,920 MAY OR MAY NOT SAY THE WORDS 8666 06:22:35,920 --> 06:22:38,160 GLYCAM WEB SO PROBABLY MORE THAN 8667 06:22:38,160 --> 06:22:39,640 A HUP PAPERS A YEAR COMING OUT 8668 06:22:39,640 --> 06:22:41,360 NOW. WHAT IS MUCH MORE 8669 06:22:41,360 --> 06:22:44,080 INTERESTING THOUGH IS ARE THE 8670 06:22:44,080 --> 06:22:47,480 SAY THE USERS OF THIS GOOI, IF 8671 06:22:47,480 --> 06:22:51,040 WE LOOK AT VISITORS WE ARE UP 8672 06:22:51,040 --> 06:22:53,760 AROUND 50,000 A YEAR. WHAT 8673 06:22:53,760 --> 06:22:57,160 CALLED UNIQUE VISITS. OR 8674 06:22:57,160 --> 06:22:59,520 VISITORS. ABOUT TWICE THAT ARE 8675 06:22:59,520 --> 06:23:00,920 UNIQUE VISITS. ABOUT A HUNDRED 8676 06:23:00,920 --> 06:23:03,440 THOUSAND A YEAR YEAR. A VISIT TWICE 8677 06:23:03,440 --> 06:23:04,680 AS MANY PER VISITOR MEANS 8678 06:23:04,680 --> 06:23:07,120 VISITOR IS DOING TWO THINGS. 8679 06:23:07,120 --> 06:23:08,200 THAT IS ONE WAY TO THINK ABOUT 8680 06:23:08,200 --> 06:23:13,880 IT. ON AVERAGE. ANOTHER METRIC 8681 06:23:13,880 --> 06:23:15,600 YOU CAN LOOK AT HOW MANY 3-D 8682 06:23:15,600 --> 06:23:16,840 STRUCTURE FILES ARE DOWNLOADED 8683 06:23:16,840 --> 06:23:18,760 EVERY YEAR. SO THEY HAVE GONE TO 8684 06:23:18,760 --> 06:23:20,680 OUR SITE NOT JUST TO LOOK ATE AT 8685 06:23:20,680 --> 06:23:22,480 IT, YOU CAN GO TO SITE AND LOOK 8686 06:23:22,480 --> 06:23:23,960 AND MODEL AND SAY THAT IS COOL 8687 06:23:23,960 --> 06:23:26,160 BUT YOU CAN DOWNLOAD PDB FILE 8688 06:23:26,160 --> 06:23:30,280 AND PLAY WITH IT WITH CHIMERA OR 8689 06:23:30,280 --> 06:23:31,600 WHATEVER YOU LIKE. NOW WE ARE UP 8690 06:23:31,600 --> 06:23:33,680 TO JUST OVER ABOUT 14,000 A 8691 06:23:33,680 --> 06:23:37,760 YEAR. THAT'S A SIGNIFICANT 8692 06:23:37,760 --> 06:23:38,880 INCREASE AND VERY SATISFYING 8693 06:23:38,880 --> 06:23:40,400 NUMBER TO SEE PEOPLE ACTUALLY 8694 06:23:40,400 --> 06:23:42,560 USE, THESE ARE NOT STATISTICS 8695 06:23:42,560 --> 06:23:45,960 THAT INCLUDE RECENT API STUFF OR 8696 06:23:45,960 --> 06:23:47,400 URL THIS IS PEOPLE GOING 8697 06:23:47,400 --> 06:23:49,360 QUAINTLY AND CLICKING AT THE EBB 8698 06:23:49,360 --> 06:23:51,320 WEBSITE. IN TERMS OF BREADTH OF 8699 06:23:51,320 --> 06:23:53,520 USE, THIS IS A SUMMARY OF THE 8700 06:23:53,520 --> 06:23:55,960 PAST FIVE YEARS. LOOKS GREAT. 8701 06:23:55,960 --> 06:23:57,960 BUT IN FACT DOESN'T LOOK MUCH 8702 06:23:57,960 --> 06:24:01,640 DIFFERENT YEAR-TO-YEAR. ROUGHLY 8703 06:24:01,640 --> 06:24:02,560 HALFWAY SPLIT BETWEEN NORTH 8704 06:24:02,560 --> 06:24:04,280 AMERICA AND EUROPE WITH QUITE A 8705 06:24:04,280 --> 06:24:05,960 BIT OF SITES IN ASIA. IT IS 8706 06:24:05,960 --> 06:24:07,240 INTERESTING IF I HAD MORE TIME I 8707 06:24:07,240 --> 06:24:08,280 WOULD SHOW THIS YEAR-TO-YEAR AND 8708 06:24:08,280 --> 06:24:10,640 YOU CAN SEE HOW IT SHIFTS AS 8709 06:24:10,640 --> 06:24:12,000 DIFFERENT GLYCOSCIENTISTS 8710 06:24:12,000 --> 06:24:14,400 DEVELOP INTEREST IN DIFFERENT 8711 06:24:14,400 --> 06:24:17,080 AREASAREAS. SOUTH AMERICA IS DIVERSE 8712 06:24:17,080 --> 06:24:19,480 AN SCATTERED. IT IS FUN. BUT IT 8713 06:24:19,480 --> 06:24:21,200 SHOWS ONE OF THE VERY EXCITING 8714 06:24:21,200 --> 06:24:23,600 THINGS ABOUT AUTONOMOUS WEB 8715 06:24:23,600 --> 06:24:26,800 TOOLS IS THEY ARE GLOBAL, NO 8716 06:24:26,800 --> 06:24:30,840 DOUBT. THIS API I MENTIONED THIS 8717 06:24:30,840 --> 06:24:34,040 TOOK AN -- AND IS ONGOING, TOOK 8718 06:24:34,040 --> 06:24:35,160 HUGE AMOUNT OF EFFORT BECAUSE 8719 06:24:35,160 --> 06:24:36,680 THE WEBSITE WASN'T DEVELOPED 8720 06:24:36,680 --> 06:24:38,440 WITH THIS IN MIND AT FIRST 8721 06:24:38,440 --> 06:24:40,520 WHATSOEVER. THE WEBSITE IS OLD. 8722 06:24:40,520 --> 06:24:42,280 AND THE FROM INFRASTRUCTURE IN 8723 06:24:42,280 --> 06:24:44,360 IF WEB DEVELOPMENT AND CODING, 8724 06:24:44,360 --> 06:24:45,400 CHANGES ALL THE TIME. AND 8725 06:24:45,400 --> 06:24:47,840 ANYBODY WHO IS DOING WEB 8726 06:24:47,840 --> 06:24:51,240 DEVELOPMENT OR HOSTING WEB TOOLS 8727 06:24:51,240 --> 06:24:53,840 FACES CHALLENGES THIS WAY 8728 06:24:53,840 --> 06:24:55,560 BECAUSE EVEN IF YOU SAY MAKE ONE 8729 06:24:55,560 --> 06:24:57,360 WAY AND KEEP IT STATIC YOU 8730 06:24:57,360 --> 06:24:59,400 CAN'T. BECAUSE THINGS LIKE 8731 06:24:59,400 --> 06:25:00,680 SECURITY PATCHES REQUIRED AND 8732 06:25:00,680 --> 06:25:02,160 THAT MAY REQUIRE UPDATE OF THIS, 8733 06:25:02,160 --> 06:25:04,560 OR THAT. SO YOU GOT TO 8734 06:25:04,560 --> 06:25:05,760 CONSTANTLY KEEP ON TOP OF STUFF. 8735 06:25:05,760 --> 06:25:09,320 WHAT WE DECIDE TO DO IS CREATE 8736 06:25:09,320 --> 06:25:11,080 PROPRIETARY SOFTWARE CALLED GEMS 8737 06:25:11,080 --> 06:25:15,160 AND GLYCAN MOLECULAR LIBRARY, 8738 06:25:15,160 --> 06:25:16,600 THAT KEEPS SCIENCE SEPARATE FROM 8739 06:25:16,600 --> 06:25:18,520 WEBSITE SO WE CAN HIRE C PLUS 8740 06:25:18,520 --> 06:25:19,960 PLUS PROGRAMMERS OR PYTHON 8741 06:25:19,960 --> 06:25:22,160 PROGRAMMERS AND THEY DON'T HAVE 8742 06:25:22,160 --> 06:25:24,720 TO LEARN CARBOHYDRATE CHEMISTRY, 8743 06:25:24,720 --> 06:25:26,160 THEY CAN BE C PLUS PLUS 8744 06:25:26,160 --> 06:25:27,280 PROGRAMMERS. LOVE TO RECRUIT 8745 06:25:27,280 --> 06:25:28,960 MORE. ALL THESE I GET SHUTTLED 8746 06:25:28,960 --> 06:25:32,280 TO VARIOUS COMPUTERS, IN MY MIND 8747 06:25:32,280 --> 06:25:36,800 IT GOES TO PARTICULAR COMPUTER 8748 06:25:36,800 --> 06:25:38,560 AND NO, NO IT GOES TO SWARM AND 8749 06:25:38,560 --> 06:25:43,320 WE FARM IT OUT. OKAY NOT MY 8750 06:25:43,320 --> 06:25:46,960 WHEEL HOUSE. WE HAVE TWO GUI'S 8751 06:25:46,960 --> 06:25:48,560 NOW, THIS IS AN ODD THING THIS 8752 06:25:48,560 --> 06:25:50,480 IS THE OLDER WITHIN WE CALL IT 8753 06:25:50,480 --> 06:25:52,040 BUILDING LEFT TO RIGHT. BECAUSE 8754 06:25:52,040 --> 06:25:58,760 YOU START ON THE GGALPNAC OR THE 8755 06:25:58,760 --> 06:26:01,760 NON-REDUCING TERMINUS OF THE 8756 06:26:01,760 --> 06:26:05,840 GRABBING TOE PYRANOSE BETA 14 8757 06:26:05,840 --> 06:26:08,200 LINKED. GAL BETA FORWARD GLCNAC 8758 06:26:08,200 --> 06:26:09,680 THAT IS HOW YOU POINT CLICK AND 8759 06:26:09,680 --> 06:26:12,760 BUILD IT. THE CHALLENGE IS WHEN 8760 06:26:12,760 --> 06:26:15,280 YOU HAVE TO BIOTYNARY WHAT 8761 06:26:15,280 --> 06:26:16,760 SHOULD I START IT DOESN'T MATTER 8762 06:26:16,760 --> 06:26:18,040 BUT THEY DON'T KNOW THAT SO THE 8763 06:26:18,040 --> 06:26:19,600 PROGRAMMERS SAID TO ME WOULDN'T 8764 06:26:19,600 --> 06:26:20,840 IT MAKE MORE SENSE TO START 8765 06:26:20,840 --> 06:26:22,560 REDUCING TERMINUS BECAUSE THERE 8766 06:26:22,560 --> 06:26:24,160 IS ONLY ONE AND BUILD THE OTHER 8767 06:26:24,160 --> 06:26:28,760 WAY? IT MIGHT MAKE MORE SENSE SO 8768 06:26:28,760 --> 06:26:30,120 THAT IS WHAT WE DID. WE HAVE A 8769 06:26:30,120 --> 06:26:31,040 DIFFERENT BUILDER HERE YOU START 8770 06:26:31,040 --> 06:26:33,680 SAYING OKAY WE HAVE MAN ATTACHED 8771 06:26:33,680 --> 06:26:36,720 TO GAL AND THEN YOU SPECIFY 8772 06:26:36,720 --> 06:26:37,800 LINKAGE THEN ANOTHER SPECIFY 8773 06:26:37,800 --> 06:26:39,760 THEN RIGHT TO LEFT. THIS BREAKS 8774 06:26:39,760 --> 06:26:42,320 MY HEAD BECAUSE I'M AN OLD 8775 06:26:42,320 --> 06:26:45,000 SCHOOL CARBOHYDRATE CHEMIST LEFT 8776 06:26:45,000 --> 06:26:46,960 TO RIGHT. BE THIS IS HOW SAY B 8777 06:26:46,960 --> 06:26:48,640 BASED CHEMISTRY IS DONE. YOU GO 8778 06:26:48,640 --> 06:26:51,640 THE OTHER WAY. THERE ARE 8779 06:26:51,640 --> 06:26:53,360 ADVANTAGES. I WOULD LIKE 8780 06:26:53,360 --> 06:26:54,520 EVERYBODY HERE TO TAKE THE TIME 8781 06:26:54,520 --> 06:26:56,400 TO GO O TO OUR SITE AND TRY LEFT 8782 06:26:56,400 --> 06:26:58,240 TO RIGHT AND RIGHT TO LEFT 8783 06:26:58,240 --> 06:26:59,680 BUILDER AND SEND ME AN EMAIL. 8784 06:26:59,680 --> 06:27:03,680 SAY I HATE THIS, I LIKE THAT. WE 8785 06:27:03,680 --> 06:27:05,600 ARE NOT SUPPORTING BOTH. ONE IS 8786 06:27:05,600 --> 06:27:08,560 GOING TO GO THE WAY OF THE DODO. 8787 06:27:08,560 --> 06:27:13,440 I HOPE IT IS NOT MINE I WILL LET 8788 06:27:13,440 --> 06:27:17,520 COMMUNITY DECIDE ON THAT. OTHER 8789 06:27:17,520 --> 06:27:20,480 THINGS WE DO, I'M PERSONALLY 8790 06:27:20,480 --> 06:27:22,360 EXCITED ABOUT IS BUILDING MODELS 8791 06:27:22,360 --> 06:27:23,760 OF GLYCOPROTEINS. WE DO THIS ALL 8792 06:27:23,760 --> 06:27:24,920 THE TIME BUT IT IS CHALLENGING 8793 06:27:24,920 --> 06:27:28,680 TO DO. I WILL SHOW A COUPLE OF 8794 06:27:28,680 --> 06:27:29,920 ISSUES. FIRST SHOW YOU WHAT WE 8795 06:27:29,920 --> 06:27:34,120 WANT TO HAVE. USER SHOULD BE 8796 06:27:34,120 --> 06:27:40,520 ABLE TO UPLOAD PBB STRUCTURE, IT 8797 06:27:40,520 --> 06:27:42,760 ALSO CAN BE THEORETICAL 8798 06:27:42,760 --> 06:27:45,160 STRUCTURE, ALPHA FOLD, A TOTALLY 8799 06:27:45,160 --> 06:27:47,000 HYPOTHETICAL HOMOLOGY MODEL. I 8800 06:27:47,000 --> 06:27:48,520 KNOW THERE'S GLYCOSYLATION OF 8801 06:27:48,520 --> 06:27:50,080 SITES OR GO TO PROGRAM THAT SAYS 8802 06:27:50,080 --> 06:27:53,200 PREDICTED GLYCOSYLATION. PUT 8803 06:27:53,200 --> 06:27:54,680 THESE GLYCANS ON AT THESE SITES 8804 06:27:54,680 --> 06:27:57,960 SO WE DO THAT. AND IT THEN 8805 06:27:57,960 --> 06:27:59,360 SCREEN RATES STATIC IMAGE OF C 8806 06:27:59,360 --> 06:28:03,160 ON THE BOTTOM BUT AND GIVES BANG 8807 06:28:03,160 --> 06:28:03,800 AND THAT IS -- WE CAN DO THAT 8808 06:28:03,800 --> 06:28:07,520 NOW. WHAT IS NICE TO DO IS HAVE 8809 06:28:07,520 --> 06:28:10,440 THIS FED INTO AUTOMATED TOOL FOR 8810 06:28:10,440 --> 06:28:11,920 MD EMULATION SO YOU GET A SENSE 8811 06:28:11,920 --> 06:28:13,160 OF HOW MUCH MOTION IS GOING ON 8812 06:28:13,160 --> 06:28:15,960 HERE. THAT IS A COSTLY THING. WE 8813 06:28:15,960 --> 06:28:19,000 WILL SEE HOW THAT GOES. THIS IS 8814 06:28:19,000 --> 06:28:20,880 NOT AS PULLY IMPLEMENTED AT THE 8815 06:28:20,880 --> 06:28:22,480 API STAGE AS WE LIKE IT YET BUT 8816 06:28:22,480 --> 06:28:24,560 IT IS GETTING THERE. SO YOU 8817 06:28:24,560 --> 06:28:27,320 WOULD PASS TO THE TOOL HERE IS 8818 06:28:27,320 --> 06:28:28,560 THE PROTEIN HERE IS 8819 06:28:28,560 --> 06:28:30,560 GLYCOSYLATION SITES, HERE IS 8820 06:28:30,560 --> 06:28:31,760 GLYCANS I WANT AT EACH SITE. ONE 8821 06:28:31,760 --> 06:28:34,000 THING THAT COMES FROM THIS, IS 8822 06:28:34,000 --> 06:28:37,160 WHEN YOU TALK THE PEOPLE WHO DO 8823 06:28:37,160 --> 06:28:39,360 GLYCOPROTEOMICS WOW IT FORCES 8824 06:28:39,360 --> 06:28:41,800 THEM TO TELL ME WHAT GLYCAN THEY 8825 06:28:41,800 --> 06:28:46,520 WANT AT EACH SITE. THAT FIRMLY 8826 06:28:46,520 --> 06:28:47,680 REINFORCES THE CONCEPTS 8827 06:28:47,680 --> 06:28:51,000 GLYCOPROTEINS ARE MIXTURES OF 8828 06:28:51,000 --> 06:28:53,440 GLYCOFORMS. YOU CAN'T HAVE 8829 06:28:53,440 --> 06:28:54,840 MIXTURE OF GLYCANSES ON SITE AND 8830 06:28:54,840 --> 06:28:56,960 MODEL IT. YOU PICK ONE AND MODEL 8831 06:28:56,960 --> 06:29:00,360 IT. THERE ARE SOME CHALLENGES 8832 06:29:00,360 --> 06:29:02,640 HERE. WHEN WE INITIALLY BUILD A 8833 06:29:02,640 --> 06:29:03,680 STRUCTURE COMPUTER DOESN'T KNOW 8834 06:29:03,680 --> 06:29:05,680 HOW TO PUT THE GLYCAN. IT'S JUST 8835 06:29:05,680 --> 06:29:07,280 A PROTEIN AND WE SAY STICK 8836 06:29:07,280 --> 06:29:09,520 GLYCAN HERE. SO WE ARBITRARILY 8837 06:29:09,520 --> 06:29:11,600 ORIENT GLYCANS ON SURFACE AND IF 8838 06:29:11,600 --> 06:29:13,120 YOU LOOK CLOSELY AT THIS YOU 8839 06:29:13,120 --> 06:29:15,760 WILL SEE THAT SOME OF THE GLYCAN 8840 06:29:15,760 --> 06:29:18,360 STRUCTURE ARE PENETRATING 8841 06:29:18,360 --> 06:29:19,760 PROTEIN SURFACE. OR BANGING INTO 8842 06:29:19,760 --> 06:29:21,760 EACH OTHER. THESE ARE WHAT WE 8843 06:29:21,760 --> 06:29:23,360 CALL ARTIFICIAL COLLISIONS OR 8844 06:29:23,360 --> 06:29:24,720 OVERLAPS, OBVIOUSLY THEY HAVE TO 8845 06:29:24,720 --> 06:29:26,560 BE RESOLVED IT IS NOT A -- IT IS 8846 06:29:26,560 --> 06:29:28,760 NOT A REAL THING. SO WE GO 8847 06:29:28,760 --> 06:29:31,080 THROUGH SOME SORT OF ALGORITHM 8848 06:29:31,080 --> 06:29:32,440 WHERE WE TWIST THESE AROUND 8849 06:29:32,440 --> 06:29:34,920 AMONG ALLOWABLE CONFIRMATIONS OF 8850 06:29:34,920 --> 06:29:37,760 THE ASPARAGINE SIDE CHAIN OR 8851 06:29:37,760 --> 06:29:39,680 ALLOWABLE INTERNAL ANGLES UNTIL 8852 06:29:39,680 --> 06:29:41,040 WE FIND SOLUTION THAT IS 8853 06:29:41,040 --> 06:29:44,680 CONSISTENT WITH EXPERIMENTAL AND 8854 06:29:44,680 --> 06:29:47,960 ENERGY COAGULATION THEN WE CAN 8855 06:29:47,960 --> 06:29:49,120 START OUR SIMULATION. THIS IS 8856 06:29:49,120 --> 06:29:50,800 REALLY EASY ON SMALL THING LIKE 8857 06:29:50,800 --> 06:29:54,160 THIS BUT IF YOU GET TO SAY GP 8858 06:29:54,160 --> 06:29:56,160 120 WITH HUGE DENSITY OF GLYCANS 8859 06:29:56,160 --> 06:29:58,760 IT BECOMES A VERY INTERESTING 8860 06:29:58,760 --> 06:30:00,080 ALGORITHMIC CHALLENGE TO FIGURE 8861 06:30:00,080 --> 06:30:01,760 THE BEST WAY TO RESOLVE THESE 8862 06:30:01,760 --> 06:30:03,960 OVERLAPS. THIS IS A FUN 8863 06:30:03,960 --> 06:30:09,480 COMPUTATIONAL PROJECT. IN TERMS 8864 06:30:09,480 --> 06:30:11,280 OF MODELING, I LIKE TO SAY WE 8865 06:30:11,280 --> 06:30:14,200 HAD A SPIKE DURING COVID-19 8866 06:30:14,200 --> 06:30:15,160 BECAUSE EVERYBODY AND THEIR 8867 06:30:15,160 --> 06:30:17,320 COLLEAGUE WAS MODELING SPIKE 8868 06:30:17,320 --> 06:30:19,400 PROTEIN. A LOT WERE USING OUR 8869 06:30:19,400 --> 06:30:21,440 TOOL YOU CAN SEE BY MONTH IN 8870 06:30:21,440 --> 06:30:24,120 APRIL OF 2020 ALMOST A THOUSAND 8871 06:30:24,120 --> 06:30:26,760 DOWNLOADS OF MODELS OF SPIKE 8872 06:30:26,760 --> 06:30:29,160 WITH GLYCANS ON IT. FEW OF THOSE 8873 06:30:29,160 --> 06:30:31,400 WERE FROM OUR GROUP AS YOU CAN 8874 06:30:31,400 --> 06:30:33,960 SEE ON THE LEFT MOSS PLOT OF 8875 06:30:33,960 --> 06:30:35,120 STRUCTURE OF THE SPIKE PROTEIN 8876 06:30:35,120 --> 06:30:36,680 WHERE WE HAVE COLORED THE 8877 06:30:36,680 --> 06:30:38,240 PROTEIN IN RED ACCORDING TO 8878 06:30:38,240 --> 06:30:40,040 ANTIBODY ACCESSIBILITY AND THEN 8879 06:30:40,040 --> 06:30:42,480 THE GLYCANS AS THE MOSS 8880 06:30:42,480 --> 06:30:45,200 STRUCTURES. WE HAVE A LOT OF 8881 06:30:45,200 --> 06:30:47,080 COLLABORATORS. THIS HAS BEEN 8882 06:30:47,080 --> 06:30:48,840 WIDELY EXTERNALLY EVALUATED 8883 06:30:48,840 --> 06:30:51,080 CROSS VALIDATED, I ENCOURAGE 8884 06:30:51,080 --> 06:30:52,640 PEOPLE TO SEND PEOPLE INTO MY 8885 06:30:52,640 --> 06:30:54,080 GROUP TO GET TRAINING RATHER 8886 06:30:54,080 --> 06:30:55,680 THAN JUST A COLLABORATOR. I LIKE 8887 06:30:55,680 --> 06:30:57,760 THE IDEA OF TEACHING SOMEONE TO 8888 06:30:57,760 --> 06:30:59,480 FISH RATHER THAN GIVING THEM A 8889 06:30:59,480 --> 06:31:01,560 FISH. AND I THINK IT PROP GATES 8890 06:31:01,560 --> 06:31:05,960 TECHNOLOGY BETTER THAT WAY. THAT 8891 06:31:05,960 --> 06:31:07,600 IS IT FOR US. THIS IS A BIG 8892 06:31:07,600 --> 06:31:10,000 TEAM PROJECT. WE HAVE WEB DEV 8893 06:31:10,000 --> 06:31:11,280 AND PYTHON DEVELOPERS C PLUS 8894 06:31:11,280 --> 06:31:15,800 PLUS PROGRAMMERS, ALL OF THIS 8895 06:31:15,800 --> 06:31:17,360 SUPPORTED MOST GRATEFULLY FROM 8896 06:31:17,360 --> 06:31:20,960 THE NIH. SO I THANK YOU GUYS FOR 8897 06:31:20,960 --> 06:31:23,080 YOUR PATIENCE AND LOOK FORWARD 8898 06:31:23,080 --> 06:31:25,040 TO SEEING YOU AT DINNER. 8899 06:31:25,040 --> 06:31:31,520 [APPLAUSE] 8900 06:31:31,520 --> 06:31:33,160 >> THANK YOU. AN ANNOUNCEMENT. 8901 06:31:33,160 --> 06:31:35,280 >> I JUST HAVE ONE BRIEF 8902 06:31:35,280 --> 06:31:37,320 ANNOUNCEMENT SO ONE OF THE 8903 06:31:37,320 --> 06:31:40,040 THINGS THAT'S BEEN SO COOL ABOUT 8904 06:31:40,040 --> 06:31:42,160 THIS MEETING IS JUST SEEING ALL 8905 06:31:42,160 --> 06:31:43,160 THE BEAUTIFUL SCIENCE AND WHERE 8906 06:31:43,160 --> 06:31:45,200 THINGS ARE GOING AND IT IS NICE 8907 06:31:45,200 --> 06:31:48,040 WE HAVE ALWAYS BEEN TALKING 8908 06:31:48,040 --> 06:31:50,480 ABOUT OUTSIDE RECOGNITION SO I 8909 06:31:50,480 --> 06:31:52,560 WANT TO JUST MENTION SOME 8910 06:31:52,560 --> 06:31:56,880 OUTSIDE RECOGNITION FOR SOMEONE 8911 06:31:56,880 --> 06:31:59,240 IN OUR AUDIENCE TODAY. WE JUST 8912 06:31:59,240 --> 06:32:01,040 FOUND OUT OR SHE JUST FOUND OUT 8913 06:32:01,040 --> 06:32:03,760 AND IT IS ALSO ONLINE THAT LINDA 8914 06:32:03,760 --> 06:32:05,360 SHAY WILSON HAS BEEN ELECTED TO 8915 06:32:05,360 --> 06:32:06,840 THE NATIONAL ACADEMY OF 8916 06:32:06,840 --> 06:32:08,400 SCIENCES. SO I THINK -- 8917 06:32:08,400 --> 06:32:14,480 [APPLAUSE] 8918 06:32:14,480 --> 06:32:17,400 WE SHOULD ALL CONGRATULATE LINDA 8919 06:32:17,400 --> 06:32:18,840 AND MAYBE SOME OF YOU WHO ARE 8920 06:32:18,840 --> 06:32:21,840 READY FOR DINNER WE CAN TOAST TO 8921 06:32:21,840 --> 00:00:00,000 HER SUCCESS. CONGRATS LINDA