1 00:00:05,920 --> 00:00:10,480 WELCOME BACK TO THE ASSAY 2 00:00:10,480 --> 00:00:12,280 WORKSHOP FOR HIGH-THROUGHPUT AND 3 00:00:12,280 --> 00:00:13,280 LEADER DISCOVERY. 4 00:00:13,280 --> 00:00:23,440 WE'RE EXCITED TO BE YOU SO, I 5 00:00:23,440 --> 00:00:25,000 KEEP REMINDING EVERYONE TO 6 00:00:25,000 --> 00:00:26,160 CONNECT WITH US SO THERE'S 7 00:00:26,160 --> 00:00:27,840 DIFFERENT WAYS YOU CAN CONNECT 8 00:00:27,840 --> 00:00:30,560 WITH US AND YOU CAN CONNECT 9 00:00:30,560 --> 00:00:35,560 THROUGH TWITTER AND USING THE 10 00:00:35,560 --> 00:00:36,600 HANDLE @ASSAY GUIDANCE. 11 00:00:36,600 --> 00:00:38,240 WE ARE ON LinkedIn SO YOU 12 00:00:38,240 --> 00:00:40,480 CAN JOIN OUR LinkedIn GROUP. 13 00:00:40,480 --> 00:00:45,040 WE POST OUR EVENTS ON NIH NCATS 14 00:00:45,040 --> 00:00:45,640 EVENTS WEBSITE AS WELL AS WE 15 00:00:45,640 --> 00:00:49,080 HAVE A WEBSITE THAT WE ARE 16 00:00:49,080 --> 00:00:51,000 ACTUALLY UPDATING SO IT WILL BE 17 00:00:51,000 --> 00:00:53,280 ONE STOP FOR YOU TO GET ALL OUR 18 00:00:53,280 --> 00:00:55,360 RESOURCES IN THE FUTURE. 19 00:00:55,360 --> 00:00:57,840 I JUST WANT TO ALSO REWIND YOU 20 00:00:57,840 --> 00:01:01,560 THAT WE HAVE AN AWESOME MANUAL, 21 00:01:01,560 --> 00:01:02,280 ELECTRONIC BOOK CALLED ASSAY 22 00:01:02,280 --> 00:01:05,480 GUIDANCE MANUAL THAT CARRIES 23 00:01:05,480 --> 00:01:07,920 MULTIPLE CHAPTERS ON ALL 24 00:01:07,920 --> 00:01:11,680 STAGES -- EARLY STAGES OF SMALL 25 00:01:11,680 --> 00:01:13,040 MOLECULE TRACK DISCOVERY AND WE 26 00:01:13,040 --> 00:01:14,400 WERE TALKING ABOUT IT ALL DAY 27 00:01:14,400 --> 00:01:15,440 YESTERDAY AND WE'LL TALK ABOUT 28 00:01:15,440 --> 00:01:16,040 IT TODAY. 29 00:01:16,040 --> 00:01:19,720 YOU CAN ACCESS THIS FOR FREE 30 00:01:19,720 --> 00:01:22,520 THROUGH NLM NCBI AND I'M SHOWING 31 00:01:22,520 --> 00:01:25,680 YOU THE LINK HERE AND AGAIN, 32 00:01:25,680 --> 00:01:27,000 WE'LL SHARE WITH YOU ALL THESE 33 00:01:27,000 --> 00:01:29,920 LINKS AND WE'LL SHARE MOST OF 34 00:01:29,920 --> 00:01:30,600 THE STRIDES AS WELL OF COURSE AT 35 00:01:30,600 --> 00:01:32,360 THE DISCRETION OF THE SPEAKERS 36 00:01:32,360 --> 00:01:37,760 AT THE END OF THE WORKSHOP. 37 00:01:37,760 --> 00:01:40,880 ONE MORE THING, WE ARE OFFERING 38 00:01:40,880 --> 00:01:42,000 DIGITAL BADGES IN COLLABORATION 39 00:01:42,000 --> 00:01:45,480 WITH THE EDUCATION BRANCH TO 40 00:01:45,480 --> 00:01:56,240 EARN A DIGITAL BADGE DESCRIBING 41 00:01:56,240 --> 00:01:57,920 HOW THE LESSONS LEARNED FROM 42 00:01:57,920 --> 00:02:00,440 THIS WORKSHOP CAN BE INTEGRATED 43 00:02:00,440 --> 00:02:01,960 AND IMPROVE YOUR RESEARCH. 44 00:02:01,960 --> 00:02:04,000 WE'LL FOLLOW YOU UP WITH 45 00:02:04,000 --> 00:02:06,080 REGARDING WITH YOU AFTER THE 46 00:02:06,080 --> 00:02:06,360 WORKSHOP. 47 00:02:06,360 --> 00:02:07,600 AS WELL. 48 00:02:07,600 --> 00:02:09,600 OK, SO, TODAY'S AGENDA, WE'RE 49 00:02:09,600 --> 00:02:12,000 REALLY EXCITED TO HAVE FANTASTIC 50 00:02:12,000 --> 00:02:17,000 TALKS TODAY AS WELL SO WE'LL 51 00:02:17,000 --> 00:02:22,240 HAVE TALKS DNA AND 3D MODELS, 52 00:02:22,240 --> 00:02:23,880 DATA ANALYSIS SO REALLY WE'LL 53 00:02:23,880 --> 00:02:28,200 COVER A LOT OF DIFFERENT SPACE 54 00:02:28,200 --> 00:02:30,560 HERE AND YOU WILL ACKNOWLEDGE 55 00:02:30,560 --> 00:02:33,440 HOW EXCITING AND INTERESTING OR 56 00:02:33,440 --> 00:02:34,720 NOT THIS TALKS ARE GOING TO BE 57 00:02:34,720 --> 00:02:37,800 AND PROVIDE A LOT OF KNOWLEDGE 58 00:02:37,800 --> 00:02:40,560 FOR YOU ALL AND HOPEFULLY HELP 59 00:02:40,560 --> 00:02:41,880 YOUR RESEARCH. 60 00:02:41,880 --> 00:02:43,200 SO, COUPLE OF THINGS WE'LL HAVE 61 00:02:43,200 --> 00:02:46,000 A BREAK AT 10:30 AND A BREAK IN 62 00:02:46,000 --> 00:02:47,480 THE AFTERNOON AT 3:00 AND WE'LL 63 00:02:47,480 --> 00:02:52,400 HAVE A LUNCH BREAK A 12:30 AND T 64 00:02:52,400 --> 00:02:53,680 5:00 WE'LL HAVE OPEN DISCUSSION 65 00:02:53,680 --> 00:02:54,040 SESSION. 66 00:02:54,040 --> 00:02:55,240 THESE ARE THE TIMES AVAILABLE 67 00:02:55,240 --> 00:02:57,480 FOR YOU TO INTERACT WITH OUR 68 00:02:57,480 --> 00:02:58,800 SPEAKERS TO TALK ABOUT YOUR 69 00:02:58,800 --> 00:03:01,640 INDIVIDUAL RESEARCH NEEDS AND 70 00:03:01,640 --> 00:03:02,680 SEEK HELP AS WELL. 71 00:03:02,680 --> 00:03:05,920 THEY'RE HERE TO TALK TO YOU SO, 72 00:03:05,920 --> 00:03:08,240 YOU KNOW, I RECOMMEND TAKING 73 00:03:08,240 --> 00:03:11,640 ADVANTAGE OF THAT OPPORTUNITY. 74 00:03:11,640 --> 00:03:13,400 SO, WITH THIS I'D LIKE TO 75 00:03:13,400 --> 00:03:18,440 INTRODUCE OUR FIRST SPEAKERS OF 76 00:03:18,440 --> 00:03:23,360 TODAY, SO Dr. TRAVIS 77 00:03:23,360 --> 00:03:25,400 MATTHEWSON AND Dr. LANE 78 00:03:25,400 --> 00:03:31,440 MILMIDED.TRAVIS WITHIN THE COMPD 79 00:03:31,440 --> 00:03:34,480 MANAGEMENT AND DISTRIBUTION AT 80 00:03:34,480 --> 00:03:36,120 PFIZER AND LANE IS A SENIOR 81 00:03:36,120 --> 00:03:38,360 SCIENTIST SUPPORTING LABORATORY 82 00:03:38,360 --> 00:03:40,520 AUTO MAKE AND LIQUID HANDLING 83 00:03:40,520 --> 00:03:41,920 WORKFLOWS AND INNSTATION FOR 84 00:03:41,920 --> 00:03:43,680 COMPOUND MANAGEMENT AND 85 00:03:43,680 --> 00:03:46,680 DISTRIBUTION AT PFIZER. 86 00:03:46,680 --> 00:03:48,560 THEIR TALK IS ENTITLED "BEST 87 00:03:48,560 --> 00:03:50,400 PRACTICES IN COMPOUND 88 00:03:50,400 --> 00:03:52,200 MANAGEMENT" AND I'M REALLY 89 00:03:52,200 --> 00:03:53,600 EXCITED TO HAVE THEM HERE TODAY. 90 00:03:53,600 --> 00:03:56,400 WELCOME TRAVIS AND LANE AND THE 91 00:03:56,400 --> 00:04:02,360 FLOOR IS YOURS. 92 00:04:02,360 --> 00:04:04,600 >>I DO WANT TO MENTION THAT 93 00:04:04,600 --> 00:04:06,800 UNFORTUNATELY, TRAVIS AND LANE 94 00:04:06,800 --> 00:04:07,920 THEY CONDITION MAKE IT TODAY SO 95 00:04:07,920 --> 00:04:09,640 THEY WILL BE PRESENTING 96 00:04:09,640 --> 00:04:13,960 VIRTUALLY. 97 00:04:13,960 --> 00:04:16,400 >>HELLO I HOPE YOU CAN HEAR ME. 98 00:04:16,400 --> 00:04:26,640 IT'S ECHOING. 99 00:04:50,720 --> 00:05:01,120 >>HANG ON JUST A SECOND. 100 00:05:08,720 --> 00:05:10,640 TRAVIS AND LANE CAN YOU HEAR ME? 101 00:05:10,640 --> 00:05:12,640 WE'RE HAVING A FEW TECHNICAL 102 00:05:12,640 --> 00:05:18,360 DIFFICULTIES, SORRY. 103 00:05:18,360 --> 00:05:19,240 I DIDN'T DO ANYTHING. 104 00:05:19,240 --> 00:05:19,960 IT WASN'T ME. 105 00:05:19,960 --> 00:05:20,400 [LAUGHTER] 106 00:05:20,400 --> 00:05:30,640 WHAT IS THAT? 107 00:05:32,600 --> 00:05:34,160 DID YOU HEAR THAT EVERYONE, IT'S 108 00:05:34,160 --> 00:05:34,400 NOT ME. 109 00:05:34,400 --> 00:05:34,800 [LAUGHTER] 110 00:05:34,800 --> 00:05:41,080 SOMETIMES IT IS ME. 111 00:05:41,080 --> 00:05:42,360 >>CAN YOU HEAR ME NOW? 112 00:05:42,360 --> 00:05:45,640 >>WE CAN HEAR YOU NOW BUT WE'RE 113 00:05:45,640 --> 00:05:47,400 WORKING ON THE VIDEO PART OF 114 00:05:47,400 --> 00:05:56,400 THIS TRANSACTION. 115 00:05:56,400 --> 00:05:58,440 THERE'S SOMETHING VERY PROFOUND 116 00:05:58,440 --> 00:06:03,800 ABOUT THIS IMAGE. 117 00:06:03,800 --> 00:06:05,600 THERE'S SOMETHING ABOUT MODERN 118 00:06:05,600 --> 00:06:15,880 TECHNOLOGY TODAY. 119 00:06:17,640 --> 00:06:19,360 WE SEE YOU AND WE HEAR YOU. 120 00:06:19,360 --> 00:06:20,760 CAN YOU SAY SOMETHING. 121 00:06:20,760 --> 00:06:20,960 GREAT. 122 00:06:20,960 --> 00:06:23,080 >>HELLO. 123 00:06:23,080 --> 00:06:27,840 GOOD MORNING. 124 00:06:27,840 --> 00:06:38,040 GOOD TO GO. 125 00:06:38,320 --> 00:06:40,120 CAN YOU HEAR ME AND SEE THE 126 00:06:40,120 --> 00:06:40,360 SLIDES. 127 00:06:40,360 --> 00:06:41,080 >>YES. 128 00:06:41,080 --> 00:06:43,560 >>OK. 129 00:06:43,560 --> 00:06:50,760 IF YOU GO INTO PRESENTATION 130 00:06:50,760 --> 00:06:56,000 MODE. 131 00:06:56,000 --> 00:06:56,600 IS THAT BETTER? 132 00:06:56,600 --> 00:06:57,520 >>NOT YET. 133 00:06:57,520 --> 00:07:02,360 >>FANTASTIC. 134 00:07:02,360 --> 00:07:02,600 NOT YET. 135 00:07:02,600 --> 00:07:13,120 NOT YET IN PRESENTATION MODE. 136 00:07:16,120 --> 00:07:18,600 YOU CAN HIT THE PRESENTATION 137 00:07:18,600 --> 00:07:20,680 ICON OR F5 WORKS IF YOU ARE ON A 138 00:07:20,680 --> 00:07:30,760 PC. 139 00:07:39,880 --> 00:07:42,320 >>ARE YOU USING A TWO-SCREEN 140 00:07:42,320 --> 00:07:42,640 SET UP? 141 00:07:42,640 --> 00:07:43,640 >>YEAH. 142 00:07:43,640 --> 00:07:48,560 LET ME JUST UNPLUG THIS. 143 00:07:48,560 --> 00:07:50,200 TECHNICAL DIFFICULTIES ALL 144 00:07:50,200 --> 00:07:50,840 AROUND. 145 00:07:50,840 --> 00:07:51,600 ARE WE THERE YET? 146 00:07:51,600 --> 00:07:52,560 IS IT WORKING RIGHT. 147 00:07:52,560 --> 00:07:57,040 >>NOT QUITE YET. 148 00:07:57,040 --> 00:07:58,480 HOW ABOUT THE ICON AT THE 149 00:07:58,480 --> 00:07:59,480 BOTTOM. 150 00:07:59,480 --> 00:08:07,520 YES, THAT ONE, EXACTLY. 151 00:08:07,520 --> 00:08:15,840 WE CAN JUST ROLL WITH THIS. 152 00:08:15,840 --> 00:08:22,440 YOU COULD ALSO TRY THE SLIDER, 153 00:08:22,440 --> 00:08:27,480 THERE, OH, NO. 154 00:08:27,480 --> 00:08:33,880 THANK YOU FOR YOUR PATIENCE, I 155 00:08:33,880 --> 00:08:35,280 APPRECIATE IT. 156 00:08:35,280 --> 00:08:42,600 TROY SHATRY SHARE AGAIN.THEN TRD 157 00:08:42,600 --> 00:08:46,360 LET'S SEE IF IT WORKS. 158 00:08:46,360 --> 00:08:46,920 NO, OK. 159 00:08:46,920 --> 00:08:47,640 ARE WE GOOD? 160 00:08:47,640 --> 00:08:49,640 >>WE'RE NOT BUT LET'S ROLL WITH 161 00:08:49,640 --> 00:08:52,920 THIS AND WE'LL GET A LITTLE 162 00:08:52,920 --> 00:08:57,560 SNEAK PREVIEW OF UPCOMING 163 00:08:57,560 --> 00:08:57,800 SLIDES. 164 00:08:57,800 --> 00:08:59,440 YOU ARE GETTING THE OTHER 165 00:08:59,440 --> 00:08:59,680 SLIDES. 166 00:08:59,680 --> 00:09:01,360 WE ONLY HAVE ONE SCREEN NOW. 167 00:09:01,360 --> 00:09:08,680 WE JUST REMOVED THE OTHER ONE. 168 00:09:08,680 --> 00:09:10,080 SORRY ABOUT THIS EVERYONE. 169 00:09:10,080 --> 00:09:16,920 SORRY WE COULDN'T BE THERE 170 00:09:16,920 --> 00:09:22,320 IN-PERSON. 171 00:09:22,320 --> 00:09:23,600 WE'LL JUST TROY WE'LL DO THE 172 00:09:23,600 --> 00:09:25,640 BEST WE CAN, SORRY ABOUT THAT. 173 00:09:25,640 --> 00:09:27,640 HI, MY NAME IS TRAVIS AND AS YOU 174 00:09:27,640 --> 00:09:29,880 HEARD I LEAD THE AUTOMATION DATA 175 00:09:29,880 --> 00:09:32,240 SUPPORT IN LIQUID FUNCTIONS FOR 176 00:09:32,240 --> 00:09:34,360 COM POUND MANAGEMENT AND I HAVE 177 00:09:34,360 --> 00:09:37,880 LANE MY AUTOMATION EXPERT AND 178 00:09:37,880 --> 00:09:39,320 WE'LL TALK ABOUT BEST PRACTICES 179 00:09:39,320 --> 00:09:41,080 FOR COMPOUND MANAGEMENT. 180 00:09:41,080 --> 00:09:42,360 SOME OF THE TOPICS WE'RE GOING 181 00:09:42,360 --> 00:09:45,440 TO COVER IN IN PRESENTATION ARE 182 00:09:45,440 --> 00:09:48,600 STORAGE CONDITIONS, COMPOUND QC 183 00:09:48,600 --> 00:09:54,480 AND JUST DATA FLOWS IN GENERAL. 184 00:09:54,480 --> 00:10:00,200 SO BACK IN 2004, WHEN MARK 185 00:10:00,200 --> 00:10:03,000 SUCKER BURG WAS COMING OUT WITH 186 00:10:03,000 --> 00:10:04,920 Facebook, THE BIGGEST KEN WAS 187 00:10:04,920 --> 00:10:06,480 THAT FREE STOP CYCLES WERE 188 00:10:06,480 --> 00:10:08,640 HAVING THE GREATEST IMPACT ON 189 00:10:08,640 --> 00:10:10,440 COMPOUND DEROGATION. 190 00:10:10,440 --> 00:10:12,440 WHAT WE FOUND INSTEAD WAS THAT A 191 00:10:12,440 --> 00:10:14,680 FRO CELL CYCLE AFTER 10 CYCLES, 192 00:10:14,680 --> 00:10:17,600 HAD VERY LITTLE IMPACT ON THE 193 00:10:17,600 --> 00:10:19,400 COMPOUNDS THEMSELVES AND 194 00:10:19,400 --> 00:10:21,720 INSTEAD, MORE IMPORTANTLY, WE 195 00:10:21,720 --> 00:10:26,480 FOUND THAT A REFRIGERATOR WAS 196 00:10:26,480 --> 00:10:27,680 THE FIRST CONDITION YOU COULD 197 00:10:27,680 --> 00:10:29,160 STORE YOUR COMPOUNDS AT. 198 00:10:29,160 --> 00:10:31,000 WE SAW THINGS WHERE BREAKING 199 00:10:31,000 --> 00:10:32,760 DOWN QUICKLY AND WE WERE GETTING 200 00:10:32,760 --> 00:10:34,320 HIGHER LEVELS OF DEGRADATION TO 201 00:10:34,320 --> 00:10:36,080 THE RECOMMENDATION AT THE TIME 202 00:10:36,080 --> 00:10:38,840 WAS KEEP THINGS AT ROOM 203 00:10:38,840 --> 00:10:40,720 TEMPERATURE FOR UP TO SIX MONTHS 204 00:10:40,720 --> 00:10:42,800 OR KEEP THEM FROZEN. 205 00:10:42,800 --> 00:10:45,400 AND IN 2019, WE DID WHAT WE 206 00:10:45,400 --> 00:10:47,560 CALLED OUR CANARY STUDY AND WHAT 207 00:10:47,560 --> 00:10:50,800 THAT WAS WAS A SELECT GROUP OF 208 00:10:50,800 --> 00:10:52,360 ABOUT 3,000 COMPOUNDS THAT WE 209 00:10:52,360 --> 00:10:53,680 BASICALLY, WE WEREN'T SO 210 00:10:53,680 --> 00:10:55,320 CONCERNED ABOUT FRO CELL CYCLES 211 00:10:55,320 --> 00:10:56,960 AT THE TIME WE WERE MORE 212 00:10:56,960 --> 00:10:58,160 CONCERNED ABOUT THE STORAGE 213 00:10:58,160 --> 00:11:08,000 CONDITIONS THEMSELVES AND FROM 214 00:11:08,000 --> 00:11:09,200 FROZEN, WE WENT TOUT A YEAR THE 215 00:11:09,200 --> 00:11:10,320 COMPOUNDS WERE PRETTY STABLE AND 216 00:11:10,320 --> 00:11:12,000 IT WAS CONSISTENT AND SO THAT IS 217 00:11:12,000 --> 00:11:15,160 WHERE WE HAD OUR RECOMMENDATIONS 218 00:11:15,160 --> 00:11:18,200 THAT IF YOU KEEP THEM AT ROOM 219 00:11:18,200 --> 00:11:19,240 TEMPERATURE, KEEP THEM IN THERE 220 00:11:19,240 --> 00:11:20,880 YOU HAVE UP TO A YEAR. 221 00:11:20,880 --> 00:11:22,680 WE USUALLY DON'T COPE THEM THOSE 222 00:11:22,680 --> 00:11:23,600 CONDITIONS FOR MORE THAN SIX 223 00:11:23,600 --> 00:11:26,440 MONTHS AND THEN THEY GO INTO 224 00:11:26,440 --> 00:11:26,760 FROZEN. 225 00:11:26,760 --> 00:11:31,080 THE REASON WE DID THIS IS 226 00:11:31,080 --> 00:11:33,240 BECAUSE -- YOU ARE STILL ON THE 227 00:11:33,240 --> 00:11:38,560 TITLE SLIDE HERE. 228 00:11:38,560 --> 00:11:48,920 I WAS ON SLIDE THREE. 229 00:11:50,880 --> 00:11:52,760 I'LL JUST SHARE MY ENTIRE 230 00:11:52,760 --> 00:11:53,320 SCREEN. 231 00:11:53,320 --> 00:12:00,520 YEAH, MAYBE. 232 00:12:00,520 --> 00:12:01,960 DO YOU SEE THE FILE? 233 00:12:01,960 --> 00:12:06,960 >>NOW WE SEE THE FILE INTEGRITY 234 00:12:06,960 --> 00:12:07,160 SLIDE. 235 00:12:07,160 --> 00:12:12,400 DO YOU -- OH YEAH! 236 00:12:12,400 --> 00:12:13,240 YEAH! 237 00:12:13,240 --> 00:12:15,880 >>NOW WE'LL FULL SCREEN AND IF 238 00:12:15,880 --> 00:12:22,720 YOU COULD JUST ROLL IT BACK A 239 00:12:22,720 --> 00:12:24,040 LITTLE BIT, MAYBE START -- HOW 240 00:12:24,040 --> 00:12:26,520 ABOUT START THERE AGAIN. 241 00:12:26,520 --> 00:12:29,040 SO BASICALLY YOU KNOW, JUST 242 00:12:29,040 --> 00:12:31,280 LETTING YOU KNOW, THESE ARE THE 243 00:12:31,280 --> 00:12:32,880 TOPICS WORE PLANNING ON 244 00:12:32,880 --> 00:12:35,160 COVERING, STORAGE, COMPOUND QC 245 00:12:35,160 --> 00:12:36,480 AND DATA FLOW THESE ARE ALL 246 00:12:36,480 --> 00:12:39,920 WORKED INTO OUR COMPOUND 247 00:12:39,920 --> 00:12:41,680 MANAGEMENT SYSTEMS. 248 00:12:41,680 --> 00:12:43,360 SO BACK TO THIS, SORRY WE 249 00:12:43,360 --> 00:12:45,240 WEREN'T ON THE SAME SLIDE. 250 00:12:45,240 --> 00:12:46,720 THE ONE WE LEFT WAS WHEN 251 00:12:46,720 --> 00:12:49,600 Facebook STARTED IN 2004 AND 252 00:12:49,600 --> 00:12:51,600 AS I WAS SAYING, AT THE TIME 253 00:12:51,600 --> 00:12:54,280 PEOPLE WERE VERY CONCERNED ABOUT 254 00:12:54,280 --> 00:12:55,840 FREE CELL CYCLES HAVING A 255 00:12:55,840 --> 00:12:58,520 SIGNIFICANT IMPACT ON COMPOUND 256 00:12:58,520 --> 00:12:59,000 DEGRADATION. 257 00:12:59,000 --> 00:13:00,280 WE FOUND FROM THIS DATA WAS THAT 258 00:13:00,280 --> 00:13:02,520 IT REALLY WAS THE STORAGE 259 00:13:02,520 --> 00:13:03,280 CONDITIONS THEM ELVES HAD A 260 00:13:03,280 --> 00:13:04,480 GREATER IMPACT THAN THE FREE 261 00:13:04,480 --> 00:13:07,200 CELL CYCLES AND WE DID 10 FREE 262 00:13:07,200 --> 00:13:10,440 CELL CYCLES, WE DID THIS FOR SIX 263 00:13:10,440 --> 00:13:11,640 MONTHS, AND THE RECOMMENDATION 264 00:13:11,640 --> 00:13:13,120 THAT CAME OUT OF THIS WAS IF YOU 265 00:13:13,120 --> 00:13:17,760 HAVE THEM AT ROOM TEMPERATURE, 266 00:13:17,760 --> 00:13:19,200 DO IT FOR NO MORE THAN SIX 267 00:13:19,200 --> 00:13:21,200 MONTHS AND TROY TO KEEP THEM 268 00:13:21,200 --> 00:13:23,200 FROZEN AND NEVER PUT ANYTHING IN 269 00:13:23,200 --> 00:13:26,440 A REFRIGERATOR AT 4C SINCE IT 270 00:13:26,440 --> 00:13:30,280 HAD THE GREATEST IMPACT ON THE 271 00:13:30,280 --> 00:13:31,720 COMPOUNDS THEMSELVES. 272 00:13:31,720 --> 00:13:33,120 IN 2019, AT THE TIME, MORE 273 00:13:33,120 --> 00:13:34,560 CONCERNED ABOUT JUST THE STORAGE 274 00:13:34,560 --> 00:13:37,080 CONDITIONS THEMSELVES, WE DID 275 00:13:37,080 --> 00:13:39,640 ANOTHER SPECIAL STUDY ON ABOUT 276 00:13:39,640 --> 00:13:41,240 3,000 COMPOUNDS AND DIVERSE SET 277 00:13:41,240 --> 00:13:43,120 OF COMPOUNDS AND THE REAL POINT 278 00:13:43,120 --> 00:13:45,760 OF THIS STUDY WAS BECAUSE OUR 279 00:13:45,760 --> 00:13:49,960 SYSTEM TEMPERATURE, HAD NO 280 00:13:49,960 --> 00:13:51,360 INDEGRADATION ISSUES AND OUR 281 00:13:51,360 --> 00:13:52,680 COMPOUNDS WERE STORED AT ROOM 282 00:13:52,680 --> 00:13:54,160 TEMPERATURE FOR GREATER THAN A 283 00:13:54,160 --> 00:13:58,880 YEAR ON OUR SYSTEM FOR ORDER 284 00:13:58,880 --> 00:13:59,200 FULFILLMENT. 285 00:13:59,200 --> 00:14:00,440 THIS GOES OUT TO A YEAR. 286 00:14:00,440 --> 00:14:02,200 WE DIDN'T SEE SIGNIFICANT 287 00:14:02,200 --> 00:14:03,080 DEGRADATION WITHIN A YEAR SO 288 00:14:03,080 --> 00:14:03,840 THAT WAS FINE. 289 00:14:03,840 --> 00:14:06,640 WE HAVE DATA THAT GOES TO -- UP 290 00:14:06,640 --> 00:14:08,320 TO TWO YEARS. 291 00:14:08,320 --> 00:14:12,680 AT LEAST WE'VE GONE BEYOND THIS. 292 00:14:12,680 --> 00:14:14,000 AGAIN, THE RECOMMENDATION IS 293 00:14:14,000 --> 00:14:16,040 DON'T PUT ANYTHING IN A 4° ROW 294 00:14:16,040 --> 00:14:17,200 FRIDGE RATER AND THAT'S PRETTY 295 00:14:17,200 --> 00:14:18,440 MUCH THE WORSE THING CAN YOU DO 296 00:14:18,440 --> 00:14:20,960 TO YOUR COMPOUNDS BUT OBVIOUSLY, 297 00:14:20,960 --> 00:14:22,520 THE BEST THING YOU CAN DO IS 298 00:14:22,520 --> 00:14:27,000 COPE KEEPTHEM FROZEN. 299 00:14:27,000 --> 00:14:28,360 IF THEY HAVE TO BE AT ROOM 300 00:14:28,360 --> 00:14:32,880 TEMPERATURE, TRY TO KEEP THEM IN 301 00:14:32,880 --> 00:14:34,640 THEIR CONDITIONS. 302 00:14:34,640 --> 00:14:36,640 WE ALSO DID AT THE TIME, SO WE 303 00:14:36,640 --> 00:14:40,640 STARTED LOOKING AT THE NEW 304 00:14:40,640 --> 00:14:41,920 ACOUSTICS TUBE TECHNOLOGY AND IN 305 00:14:41,920 --> 00:14:43,000 THESE TUBES WE WANTED TO KNOW 306 00:14:43,000 --> 00:14:45,640 HOW FAST IS THE DMSO ABSORBING 307 00:14:45,640 --> 00:14:49,640 WATER AND WE KNOW DMSO IS HYDRO 308 00:14:49,640 --> 00:14:51,040 SCOPIC AND THE ONE ON THE LEFT, 309 00:14:51,040 --> 00:14:52,960 THAT WAS OVER THE COURSE A 310 00:14:52,960 --> 00:14:55,600 REGULAR BUSINESS DAY SO YOU CAN 311 00:14:55,600 --> 00:14:57,240 SEE EVEN IN JUST A REGULAR 312 00:14:57,240 --> 00:14:58,360 EIGHT-HOUR DAY YOU WOULD GET 313 00:14:58,360 --> 00:15:01,800 CLOSE TO EQUILIBRIUM IF THEY'RE 314 00:15:01,800 --> 00:15:02,960 EXPOSED IN THE LAB CONDITIONS 315 00:15:02,960 --> 00:15:11,120 FOR THAT TIME. 316 00:15:11,120 --> 00:15:13,000 THE ONE ON THE RIGHT WE STRETCH 317 00:15:13,000 --> 00:15:14,280 OUT BECAUSE THAT DRAMATIC LINE 318 00:15:14,280 --> 00:15:16,320 IN THE LEFT WAS OVERNIGHT SO 319 00:15:16,320 --> 00:15:17,520 WHAT YOU CAN SEE THERE IS THAT 320 00:15:17,520 --> 00:15:28,000 AS YOU APPROACH EQUILIBRIUM. 321 00:15:28,560 --> 00:15:32,800 THE REAL POINT, EVEN IN THESE 322 00:15:32,800 --> 00:15:34,000 ACOUSTIC TUBES WITH A SMALL 323 00:15:34,000 --> 00:15:37,040 CENSUS AREA, 100% DSMO IS 324 00:15:37,040 --> 00:15:40,240 ABSORBING WATER AT A VERY RAPID 325 00:15:40,240 --> 00:15:40,840 RATE. 326 00:15:40,840 --> 00:15:45,360 SO JUST SOME GENERAL THOUGHTS 327 00:15:45,360 --> 00:15:48,280 FOR BEST PRACTICES IF YOU ARE AT 328 00:15:48,280 --> 00:15:50,280 ROOM TEMPERATURE THREE MONTHS 329 00:15:50,280 --> 00:15:52,200 DON'T PUT ANYTHING AT 4C WE GO 330 00:15:52,200 --> 00:15:53,920 UP TO SIX MONTHS AT ROOM 331 00:15:53,920 --> 00:15:55,680 TEMPERATURE AND THAT IS BEEN 332 00:15:55,680 --> 00:16:00,600 PRETTY GOOD FOR OUR COMPOUNDS. 333 00:16:00,600 --> 00:16:06,880 KEEP THEM DRIVEWAY THEM DRY. 334 00:16:06,880 --> 00:16:08,800 IF LONG-TERM STORAGE MAKE SURE 335 00:16:08,800 --> 00:16:10,720 THEY'RE SEALED AND FROZEN -20. 336 00:16:10,720 --> 00:16:11,920 NEVER, NEVER AT 4°. 337 00:16:11,920 --> 00:16:13,600 ALL THE IMAGES ON THE RIGHT SIDE 338 00:16:13,600 --> 00:16:15,840 THERE, BASICALLY REFLECT WHAT 339 00:16:15,840 --> 00:16:19,320 HAPPENS WITH DMSO, TRY TO ALWAYS 340 00:16:19,320 --> 00:16:21,920 DEEP YOUR DILUTION AT DMSO FOR 341 00:16:21,920 --> 00:16:28,480 100% AND GO A KEY US AND YOU 342 00:16:28,480 --> 00:16:31,240 NEED TO USE IT WITHIN 24 HOUR 343 00:16:31,240 --> 00:16:34,960 AND LEAVE THOSE HANGING AROUND 344 00:16:34,960 --> 00:16:37,760 AQUEOUS FOR LONGER THAN THAT. 345 00:16:37,760 --> 00:16:44,840 TO REITERATE SOME CONSIDERATIONS 346 00:16:44,840 --> 00:16:54,960 HERE. 347 00:16:56,520 --> 00:16:58,680 WE HAD QC FAIL AND THIS IS THE 348 00:16:58,680 --> 00:16:59,640 CASE ACROSS THE BOARD. 349 00:16:59,640 --> 00:17:01,320 YOU WILL NEVER HAVE A PERFECT 350 00:17:01,320 --> 00:17:01,520 FILE. 351 00:17:01,520 --> 00:17:03,960 WE EXPECT TO ROUND 10% TO 15% 352 00:17:03,960 --> 00:17:06,200 THAT HAS SOME SORT OF ISSUE. 353 00:17:06,200 --> 00:17:07,760 SO JUST BE VERY FAMILIAR WITH 354 00:17:07,760 --> 00:17:10,040 THAT THAT THAT'S WHAT YOU ARE 355 00:17:10,040 --> 00:17:10,880 DEALING WITH. 356 00:17:10,880 --> 00:17:12,440 YOU SHOULD ALWAYS TRY TO WHERE 357 00:17:12,440 --> 00:17:13,720 YOUR QUALITY OF YOUR COMPOUNDS 358 00:17:13,720 --> 00:17:15,840 AND THE STEWARDSHIP ON YOUR 359 00:17:15,840 --> 00:17:17,280 COMPOUNDS AND IF YOU HAVE ANY 360 00:17:17,280 --> 00:17:18,720 KIND OF COM POUND QUALITY ISSUES 361 00:17:18,720 --> 00:17:22,240 THAT COULD INTERFERE WITH THE 362 00:17:22,240 --> 00:17:24,800 QUALITY OF THE COMPASS FOR ASSAY 363 00:17:24,800 --> 00:17:26,120 IT'S GOOD TO KNOW AND UTILIZE 364 00:17:26,120 --> 00:17:27,680 YOUR RESOURCES SO YOU REALLY 365 00:17:27,680 --> 00:17:29,560 UNDERSTAND WHAT YOU ARE WORKING 366 00:17:29,560 --> 00:17:30,040 WITH. 367 00:17:30,040 --> 00:17:32,680 MAKE SURE YOU ARE ALWAYS VERY 368 00:17:32,680 --> 00:17:32,920 PROACTIVE. 369 00:17:32,920 --> 00:17:34,120 YOU UNDERSTAND THAT THE ASSAY 370 00:17:34,120 --> 00:17:35,720 AND ANY SENSITIVITY TO ANY 371 00:17:35,720 --> 00:17:38,720 COMPOUND QUALITY ISSUES IS 372 00:17:38,720 --> 00:17:39,120 IMPORTANT. 373 00:17:39,120 --> 00:17:41,120 IF YOU HAVE TO DO A RISK 374 00:17:41,120 --> 00:17:42,320 ASSESSMENT AS FAR AS THE 375 00:17:42,320 --> 00:17:44,520 QUALITY, INTEGRITY AND MAYBE YOU 376 00:17:44,520 --> 00:17:46,520 NEED TO DO A DEEPER 377 00:17:46,520 --> 00:17:47,200 CHARACTERIZATION OF THE 378 00:17:47,200 --> 00:17:48,280 COMPOUNDS SO SEE WHAT THEY'RE 379 00:17:48,280 --> 00:17:52,000 ACTUALLY DOING, WHAT WE 380 00:17:52,000 --> 00:17:53,440 BASICALLY ARE FROM DRY TO 381 00:17:53,440 --> 00:17:55,840 WHATEVER CONCENTRATION YOU ARE 382 00:17:55,840 --> 00:17:57,240 STORING AT LIQUID, THAT'S WERE 383 00:17:57,240 --> 00:17:59,400 THE BIGGEST ERROR TIMES COME IN 384 00:17:59,400 --> 00:18:03,080 SO THEY'RE NOMINAL 385 00:18:03,080 --> 00:18:03,400 CONCENTRATION. 386 00:18:03,400 --> 00:18:06,360 IT'S PLUS OR MINUS. 387 00:18:06,360 --> 00:18:09,320 JUST TO GO INTO OUR QC PROCESS, 388 00:18:09,320 --> 00:18:12,080 SO WE HAVE OUR AUTOMATED QC 389 00:18:12,080 --> 00:18:13,560 PROCESS. 390 00:18:13,560 --> 00:18:14,720 FOR OUR NEELY SYNTHESIZED 391 00:18:14,720 --> 00:18:16,040 COMPOUNDS WE HAVE OUR PHRASES, 392 00:18:16,040 --> 00:18:18,880 IF YOU SCREEN IT WE'LL QC IT. 393 00:18:18,880 --> 00:18:21,200 WE HAVE A 30-DAY WINDOW IN WHICH 394 00:18:21,200 --> 00:18:23,240 A COMPOUND IF IT GETS ORDERED IT 395 00:18:23,240 --> 00:18:26,720 WILL GO TO QC UNLESS THERE'S A 396 00:18:26,720 --> 00:18:27,520 QC VALUE PRESENT. 397 00:18:27,520 --> 00:18:29,440 WE USE THIS CRITERIA AND WE HAVE 398 00:18:29,440 --> 00:18:31,840 A PASS-FAIL SUSPECT RESULT TO 399 00:18:31,840 --> 00:18:33,080 EACH OF THE COMPOUNDS AND WE 400 00:18:33,080 --> 00:18:35,480 ALSO CHECK TO MAKE SURE THAT THE 401 00:18:35,480 --> 00:18:37,280 MASS WAS FOUND AND THAT THE 402 00:18:37,280 --> 00:18:39,120 PURITY IS GREATER THAN 80%. 403 00:18:39,120 --> 00:18:41,680 WE HAVE THIS CATCH-ALL CATEGORY 404 00:18:41,680 --> 00:18:42,840 CALLED SUSPECT WHICH CAUSES 405 00:18:42,840 --> 00:18:44,640 CONFUSION AND BASICALLY THAT IS 406 00:18:44,640 --> 00:18:45,240 EXACTLY THAT. 407 00:18:45,240 --> 00:18:47,480 IT'S A CATCH-ALL. 408 00:18:47,480 --> 00:18:49,320 THE METHOD FAILED. 409 00:18:49,320 --> 00:18:50,240 THE INSTRUMENT FAILED. 410 00:18:50,240 --> 00:18:51,080 IT COULD BE ANYTHING. 411 00:18:51,080 --> 00:18:52,840 I TROY TO TELL PEOPLE IF YOU SEE 412 00:18:52,840 --> 00:18:55,000 A SUSPECT, ROW SEND IT FOR QC OR 413 00:18:55,000 --> 00:18:56,880 IT JUST DOESN'T WORK IN OUR 414 00:18:56,880 --> 00:18:59,440 METHOD AND YOU WON'T GET A QC 415 00:18:59,440 --> 00:19:00,600 AND SOMETHING ELSE HAS TO BE 416 00:19:00,600 --> 00:19:00,840 DONE. 417 00:19:00,840 --> 00:19:03,200 IT HAS THE CONNOTATION OF IT'S 418 00:19:03,200 --> 00:19:04,240 SUSPECT THERE'S SOMETHING WRONG. 419 00:19:04,240 --> 00:19:06,040 IT COULD BE FINE. 420 00:19:06,040 --> 00:19:09,200 AND THEN, I WANTED TO JUST POINT 421 00:19:09,200 --> 00:19:12,480 OUT, IT'S UV215 HAS TO BE 80% OR 422 00:19:12,480 --> 00:19:12,720 GREATER. 423 00:19:12,720 --> 00:19:13,920 THIS IS A HARD LINE. 424 00:19:13,920 --> 00:19:15,200 IT'S GOT TO UNDERSTAND THAT DATA 425 00:19:15,200 --> 00:19:16,640 BECAUSE IT'S VERY POSSIBLE TO 426 00:19:16,640 --> 00:19:19,520 GET A COMPOUND THAT WILL BE 79 427 00:19:19,520 --> 00:19:20,960 AND A HALF, THE FIRST TIME YOU 428 00:19:20,960 --> 00:19:23,360 RUN IT AND 80 THE NEXT TIME AND 429 00:19:23,360 --> 00:19:24,640 YES, THE FIRST ONE WILL BE 430 00:19:24,640 --> 00:19:27,440 LABELED A FAIL AND THE NEXT 431 00:19:27,440 --> 00:19:28,200 RESULT WILL BE LABELED A PASS. 432 00:19:28,200 --> 00:19:29,480 IT'S IMPORTANT TO UNDERSTAND HOW 433 00:19:29,480 --> 00:19:31,760 THAT DATA WORKS AND HOW TO APPLY 434 00:19:31,760 --> 00:19:35,520 THAT TO THE COMPOUNDS THAT WE'RE 435 00:19:35,520 --> 00:19:36,320 LOOKING AT. 436 00:19:36,320 --> 00:19:39,160 THERE'S AN EXAMPLE, SO, ON THE 437 00:19:39,160 --> 00:19:40,600 TOP, IT'S PROBABLY HARD TO SEE 438 00:19:40,600 --> 00:19:43,280 BUT BASICALLY WE HAVE A SYSTEM 439 00:19:43,280 --> 00:19:45,240 SET UP, WHERE WHEN WE DELIVER 440 00:19:45,240 --> 00:19:47,160 PLATES TO OUR CUSTOMERS, OUR 441 00:19:47,160 --> 00:19:48,120 SCREENERS, THEY HAVE THE TOOL 442 00:19:48,120 --> 00:19:52,000 THEY CAN CLICK ON AND SCAN THE 443 00:19:52,000 --> 00:19:53,880 BAR CODE AND IT WILL GIVE THEM A 444 00:19:53,880 --> 00:19:55,000 PICTURE OF THEIR PLATE AND ANY 445 00:19:55,000 --> 00:19:57,720 OF THE MOST RECENT QC DATA ON 446 00:19:57,720 --> 00:19:58,160 THE TOP. 447 00:19:58,160 --> 00:20:00,560 YOU CAN SEE YOUR PASS, FAILS AND 448 00:20:00,560 --> 00:20:01,080 SUSPECTS. 449 00:20:01,080 --> 00:20:02,280 IF THERE'S NO VALUE THERE, THE 450 00:20:02,280 --> 00:20:03,960 RESULT HASN'T BEEN PUBLISHED OR 451 00:20:03,960 --> 00:20:05,600 RUN YET SO MAYBE THEY NEED TO 452 00:20:05,600 --> 00:20:09,760 GET IT ROW RUN AND THEN IN THE 453 00:20:09,760 --> 00:20:11,640 BOTTOM, WE HAVE PRECIPITATE 454 00:20:11,640 --> 00:20:12,440 DATA. 455 00:20:12,440 --> 00:20:14,240 WE USE THIS TWO BORDER WE HAVE 456 00:20:14,240 --> 00:20:17,080 AND IT TAKES A PICTURE OF THE 457 00:20:17,080 --> 00:20:22,160 COMPOUNDS SOL R IN TUBES AND IT 458 00:20:22,160 --> 00:20:24,520 DETERMINES IF IT THIS IS THERE'S 459 00:20:24,520 --> 00:20:27,760 MY PRECIPITATE IN THE TUBES. 460 00:20:27,760 --> 00:20:29,560 WE DON'T PUBLISH A NO 461 00:20:29,560 --> 00:20:29,920 INTENTIONALLY. 462 00:20:29,920 --> 00:20:31,360 WE ONLY PUBLISH A YES AND IT'S 463 00:20:31,360 --> 00:20:35,800 NOT A YES IT'S PRECIPITATE YOU 464 00:20:35,800 --> 00:20:38,200 SHOULD BE QUESTIONING IF YOU SEE 465 00:20:38,200 --> 00:20:39,000 A YES. 466 00:20:39,000 --> 00:20:40,440 WE HAVE A PROCEDURE WHERE IF 467 00:20:40,440 --> 00:20:41,840 SOMEONE REALLY WANTS 468 00:20:41,840 --> 00:20:44,320 CONFIRMATION, WE CAN ACTUALLY GO 469 00:20:44,320 --> 00:20:46,080 PULL THE SOURCE TUBE AND LOOK AT 470 00:20:46,080 --> 00:20:49,200 IT AND MICK A JUDGMENT CALL. 471 00:20:49,200 --> 00:20:50,680 THIS GIVES EVERYBODY RAPID 472 00:20:50,680 --> 00:20:54,000 ACCESS TO ALL THE DATA THAT THEY 473 00:20:54,000 --> 00:20:56,720 CAN JUST MARRY UP TO WHAT THEY 474 00:20:56,720 --> 00:20:58,160 HAVE AND HAVE THIS INFORMATION 475 00:20:58,160 --> 00:21:05,280 PREPARED WHEN THEY DO THE 476 00:21:05,280 --> 00:21:05,760 SCREENING. 477 00:21:05,760 --> 00:21:08,200 SO WHEN THINKING ABOUT KIND OF 478 00:21:08,200 --> 00:21:10,840 TRIAGE AND COMPOUND QC ONE THING 479 00:21:10,840 --> 00:21:12,600 IS IMPORTANT TO BE AWARE THERE 480 00:21:12,600 --> 00:21:14,920 ARE BATCH-TO-BATCH DIFFERENCES 481 00:21:14,920 --> 00:21:16,240 SO WE HAVE SEVERAL BATCHES OF 482 00:21:16,240 --> 00:21:17,640 OUR COMPOUNDS AND OFTEN TIMES 483 00:21:17,640 --> 00:21:20,400 ORDERS COME IN FROM A PARENT 484 00:21:20,400 --> 00:21:21,880 STRUCTURE AND NO ONE CARES ABOUT 485 00:21:21,880 --> 00:21:23,120 WHAT BATCHES AT THE INITIAL TIME 486 00:21:23,120 --> 00:21:24,960 FOR A HIGH SCREEN PER SE BUT 487 00:21:24,960 --> 00:21:26,720 IT'S IMPORTANT TO KNOW THERE ARE 488 00:21:26,720 --> 00:21:30,000 OTHER BATCHES SO IF YOU ARE 489 00:21:30,000 --> 00:21:31,160 GETTING SOME STRANGE DATA, 490 00:21:31,160 --> 00:21:32,800 THERE'S ALWAYS AN OPPORTUNITY TO 491 00:21:32,800 --> 00:21:35,440 MAYBE GO FIND AN ALTERNATE BATCH 492 00:21:35,440 --> 00:21:38,360 AND ALWAYS CHECK COMPOUND QC, IF 493 00:21:38,360 --> 00:21:41,240 THERE ISN'T A RESENT QC DATA 494 00:21:41,240 --> 00:21:43,040 VALUE IT'S POSSIBLE TO SEND IT 495 00:21:43,040 --> 00:21:45,800 TO YOUR QC SCREEN AND GET AN 496 00:21:45,800 --> 00:21:46,840 UPDATED VALUE ON THE COMPOUNDS 497 00:21:46,840 --> 00:21:48,000 YOU ARE RUNNING SO YOU CAN TAKE 498 00:21:48,000 --> 00:21:49,640 A LOOK AT THAT AND IT'S 499 00:21:49,640 --> 00:21:51,840 IMPORTANT TO PRETTY MUCH CHECK 500 00:21:51,840 --> 00:21:53,720 EVERY OPTION YOU HAVE BEFORE YOU 501 00:21:53,720 --> 00:21:55,760 TRY TO COMMIT SYNTHETIC CHEM IS 502 00:21:55,760 --> 00:21:57,160 TREMENDOUS RESOURCES TO HAVE IT 503 00:21:57,160 --> 00:21:57,680 THE SIZE. 504 00:21:57,680 --> 00:21:58,760 THIS IS VERY IMPORTANT. 505 00:21:58,760 --> 00:22:03,520 I MEAN, I KNOW IT'S THE LAST 506 00:22:03,520 --> 00:22:05,440 RESORT FOR US WE LOOK AT WHAT IS 507 00:22:05,440 --> 00:22:06,960 AVAILABLE AND DUE AND WHAT OTHER 508 00:22:06,960 --> 00:22:13,040 SOURCES OF THE COMPOUND ARE IN 509 00:22:13,040 --> 00:22:13,400 THERE. 510 00:22:13,400 --> 00:22:14,960 SO PULLING THIS TOGETHER AND 511 00:22:14,960 --> 00:22:16,840 APPLYING TO YOUR HTS SCREENING 512 00:22:16,840 --> 00:22:18,480 FUNNEL AND IT'S VERY STANDARD. 513 00:22:18,480 --> 00:22:20,840 YOU RUN YOUR PRIMARY HTS AND 514 00:22:20,840 --> 00:22:23,680 KEEP YOUR STOCKS AT 100% DMSO 515 00:22:23,680 --> 00:22:26,400 AND DON'T GO AQUEOUS AND RUN 516 00:22:26,400 --> 00:22:27,680 THEM WITHIN THAT DAY IT'S 517 00:22:27,680 --> 00:22:29,480 IMPORTANT AND WHEN YOU GET YOUR 518 00:22:29,480 --> 00:22:31,760 HITS BACK, IF YOU, FOR WHATEVER 519 00:22:31,760 --> 00:22:33,840 REASON, YOU HAVE AN UNEXPECTED 520 00:22:33,840 --> 00:22:39,640 LILYLOW RATE, RETEST BUT MAYBE U 521 00:22:39,640 --> 00:22:42,080 SHOULD LOOK FOR YOUR ANALYTICAL 522 00:22:42,080 --> 00:22:43,800 DATA AND USE THAT AS PART OF 523 00:22:43,800 --> 00:22:47,200 YOUR TRIAGE TO SEE IF THE QC 524 00:22:47,200 --> 00:22:48,680 DATA WAS BAD OR THERE'S 525 00:22:48,680 --> 00:22:50,360 SOMETHING AN ALTERNATE BATCH YOU 526 00:22:50,360 --> 00:22:53,280 NEED TO LOOK AT AND THEN, AS YOU 527 00:22:53,280 --> 00:22:55,320 MOVE INTO YOUR FOLLOW-UP AND 528 00:22:55,320 --> 00:22:58,160 YOUR IC50s, AGAIN, SAME THING, 529 00:22:58,160 --> 00:23:00,600 DON'T PREPARE YOUR IC50s 530 00:23:00,600 --> 00:23:03,560 AQUEOUS KEEP EVERYTHING AT 100% 531 00:23:03,560 --> 00:23:05,560 DMSO AND GO INTO YOUR ASSAY 532 00:23:05,560 --> 00:23:07,320 PLATE BEFORE YOU DELAWSUIT 533 00:23:07,320 --> 00:23:08,720 AQUEOUS AND RUN THEM THE SAME 534 00:23:08,720 --> 00:23:09,200 DAY. 535 00:23:09,200 --> 00:23:11,200 AGAIN, IT'S IMPORTANT TO TAKE A 536 00:23:11,200 --> 00:23:13,360 LOOK AT ANY ANALYTICAL 537 00:23:13,360 --> 00:23:14,080 CHARACTERIZATION DATA THAT YOU 538 00:23:14,080 --> 00:23:15,280 HAVE AND ININCLUDED THAT AS PART 539 00:23:15,280 --> 00:23:18,160 OF YOUR FILTERING FOR YOUR HITS 540 00:23:18,160 --> 00:23:21,000 AND THEN VERIFY EVERYTHING 541 00:23:21,000 --> 00:23:25,880 BEFORE GOING BACK TO RESENT 542 00:23:25,880 --> 00:23:26,360 ASSISTS. 543 00:23:26,360 --> 00:23:29,800 SO AGAIN, BEST PRACTICES, 544 00:23:29,800 --> 00:23:32,200 ESPECIALLY FOR -- NEVER USE OLD 545 00:23:32,200 --> 00:23:33,680 DMSO UNDER LAB CONDITIONS YOU 546 00:23:33,680 --> 00:23:36,760 WILL GET THE EQUILIBRIUM 547 00:23:36,760 --> 00:23:37,240 QUICKLY. 548 00:23:37,240 --> 00:23:39,600 DON'T LEAVE THE CAP OFF THE DMSO 549 00:23:39,600 --> 00:23:41,320 DO LEAVE IT OUT AND DON'T TRUST 550 00:23:41,320 --> 00:23:43,520 IT SITTING ON THE DECK ALL DAY. 551 00:23:43,520 --> 00:23:45,240 MAKE SURE YOU SWAP THOSE OUT. 552 00:23:45,240 --> 00:23:49,000 KEEPING IT FRESH. 553 00:23:49,000 --> 00:23:50,560 ONE OF OUR RECOMMENDATIONS WHEN 554 00:23:50,560 --> 00:23:53,320 WE GET DRY, WE RECOMMEND NOT 555 00:23:53,320 --> 00:23:56,200 ORDERING LESS THAN TWO MIGS OF A 556 00:23:56,200 --> 00:23:58,960 SAMPLE AT A TIME BECAUSE THE 557 00:23:58,960 --> 00:24:00,840 VARIATION IN CALIBRATION. 558 00:24:00,840 --> 00:24:02,400 IT'S WHERE THE BIGGEST ERROR 559 00:24:02,400 --> 00:24:10,760 COMES IN IF THEY ASK FOR ONE 560 00:24:10,760 --> 00:24:13,320 MIG, YOU CAN HAVE VARIATION AND 561 00:24:13,320 --> 00:24:15,040 YOU WILL BE OFF. 562 00:24:15,040 --> 00:24:16,520 IT'S ALSO IMPORTANT TO CHECK 563 00:24:16,520 --> 00:24:18,080 YOUR LIQUID HANDLING EQUIPMENT 564 00:24:18,080 --> 00:24:19,400 AND MAKE SURE THAT YOU ARE 565 00:24:19,400 --> 00:24:20,760 FOLLOWING ALL THE PROPER 566 00:24:20,760 --> 00:24:22,680 MAINTENANCE, REGULAR 567 00:24:22,680 --> 00:24:24,680 MAINTENANCE, GETTING PMs DONE 568 00:24:24,680 --> 00:24:27,680 AND MAKING SURE THEY'RE 569 00:24:27,680 --> 00:24:29,240 CALIBRATED AND YOU QC THEM AS 570 00:24:29,240 --> 00:24:29,800 WELL. 571 00:24:29,800 --> 00:24:31,080 SO, YOU HEARD ME TALK BEFORE 572 00:24:31,080 --> 00:24:33,240 ABOUT THE ACOUSTIC TUBES. 573 00:24:33,240 --> 00:24:35,080 THIS IS THE CUTTING EDGE 574 00:24:35,080 --> 00:24:36,160 TECHNOLOGY FOR COMPOUND 575 00:24:36,160 --> 00:24:37,400 MANAGEMENT STORAGE. 576 00:24:37,400 --> 00:24:39,480 WE HAVE THE MIDDLE, YOU CAN SEE 577 00:24:39,480 --> 00:24:41,880 THIS BIG PICTURE THAT'S A LARGE 578 00:24:41,880 --> 00:24:43,000 SAMPLE STORE WE HAVE THAT WE 579 00:24:43,000 --> 00:24:45,960 HOLD ALL OF OUR TUBES AND IN 580 00:24:45,960 --> 00:24:48,360 THERE ARE THESE VERY SMALL 581 00:24:48,360 --> 00:24:48,720 ACOUSTIC TUBES. 582 00:24:48,720 --> 00:24:49,760 THEY'RE VERY INTERESTING, THEY 583 00:24:49,760 --> 00:24:53,160 ALL HAVE INDIVIDUALLY SEALS WITH 584 00:24:53,160 --> 00:24:53,760 SCREW CAP. 585 00:24:53,760 --> 00:24:55,960 YOU WILL SEE THESE FOUR TINY BAR 586 00:24:55,960 --> 00:24:57,600 CODES ON THE BOTTOM AND THIS IS 587 00:24:57,600 --> 00:24:58,440 CLEVER WHAT THEY DID. 588 00:24:58,440 --> 00:25:00,600 THEY FOUND THAT THE BAR CODES 589 00:25:00,600 --> 00:25:03,280 ITSELF ON THE BOTTOM INDER 590 00:25:03,280 --> 00:25:04,960 FEARED WITH THE ACOUSTIC 591 00:25:04,960 --> 00:25:05,400 TECHNOLOGY. 592 00:25:05,400 --> 00:25:06,360 THERE'S A DIAGRAM OF HOW IT 593 00:25:06,360 --> 00:25:07,320 WORKS OVER HERE. 594 00:25:07,320 --> 00:25:10,400 IT ACTUALLY INTERFERED WITH THE 595 00:25:10,400 --> 00:25:12,280 ACOUSTIC SO THEY SPLIT IT NO 596 00:25:12,280 --> 00:25:13,760 FOUR. 597 00:25:13,760 --> 00:25:14,480 WHY FOUR? 598 00:25:14,480 --> 00:25:16,680 IT'S DOUBLE REDUNDANT. 599 00:25:16,680 --> 00:25:19,080 IT'S TWO HALF OF THE BAR CUED 600 00:25:19,080 --> 00:25:20,120 SPLIT IN COUPE LA INDICATE SO 601 00:25:20,120 --> 00:25:22,480 EVEN IF DAMAGE HAPPENS TO TWO OF 602 00:25:22,480 --> 00:25:24,160 THESE, AS LONG AS THEY'RE NOT ON 603 00:25:24,160 --> 00:25:26,720 THE DIAGONAL IT CAN READ THE BAR 604 00:25:26,720 --> 00:25:28,640 CODE WITHOUT ISSUES. 605 00:25:28,640 --> 00:25:31,880 THESE ARE ALL INTER 96 RACK. 606 00:25:31,880 --> 00:25:35,120 SO WE CAN PUT SIX OF THESE RACKS 607 00:25:35,120 --> 00:25:35,400 ON A TRAY. 608 00:25:35,400 --> 00:25:37,800 THEY GET LOADED AND RE FORMATTED 609 00:25:37,800 --> 00:25:39,840 INTO A D TRAY AND IT'S JUST TO 610 00:25:39,840 --> 00:25:41,480 INCREASE THE STORAGE INSIDE THE 611 00:25:41,480 --> 00:25:41,720 STORAGE. 612 00:25:41,720 --> 00:25:43,720 BY DOING THAT IN THE SAME SPACE 613 00:25:43,720 --> 00:25:47,000 THAT WE HAVE SIX RACKS, WE'RE 614 00:25:47,000 --> 00:25:49,480 FITTING 1,000 AND 8 TUBES IN THE 615 00:25:49,480 --> 00:25:50,760 HC RACK SO THIS PICTURE IS OUR 616 00:25:50,760 --> 00:25:54,000 NEW SYSTEM THAT WE USE WITH THE 617 00:25:54,000 --> 00:25:56,040 NEW ECHOES, SUCCESS IT THAT CAN 618 00:25:56,040 --> 00:26:00,440 USE THESE AS A SOURCE OR A 619 00:26:00,440 --> 00:26:04,320 COMPOUND AND THE TUBES AND IT 620 00:26:04,320 --> 00:26:06,120 KEEPS ALL SAMPLES FROZEN AND NOT 621 00:26:06,120 --> 00:26:09,400 IN USE AND ONLY EXPOS SAMPLES TO 622 00:26:09,400 --> 00:26:10,600 SIGN AND ROOM TEMPERATURE 623 00:26:10,600 --> 00:26:11,440 CONDITIONS WHEN THEY'RE BEING 624 00:26:11,440 --> 00:26:13,440 USED TO FULFILL AN ORDER AND THE 625 00:26:13,440 --> 00:26:16,200 ECHO SO THEY GO IN THE ECHO IT 626 00:26:16,200 --> 00:26:19,640 DETECTS THE BOTTOM AND EJECTS 627 00:26:19,640 --> 00:26:21,200 THE 2.5 DROPS INTO THE TARGET 628 00:26:21,200 --> 00:26:22,160 PLATES. 629 00:26:22,160 --> 00:26:23,400 COMPARED TA OUR OLD SYSTEM WE 630 00:26:23,400 --> 00:26:25,600 HAD TO DO OUR CANARY STUDY 631 00:26:25,600 --> 00:26:27,280 BECAUSE OUR PLATES BASICALLY SIT 632 00:26:27,280 --> 00:26:30,680 AT ROOM TEMPERATURE OUT ON RACKS 633 00:26:30,680 --> 00:26:31,960 AND AGAIN, IT'S BECAUSE THEY'RE 634 00:26:31,960 --> 00:26:33,480 IN PLATES IF YOU ORDER ONE 635 00:26:33,480 --> 00:26:35,920 SAMPLE FROM A PLATE, IT HAS TO 636 00:26:35,920 --> 00:26:36,400 BE UNSEALED. 637 00:26:36,400 --> 00:26:38,360 THE WHOLE PLATE OF COMPOUNDS IS 638 00:26:38,360 --> 00:26:39,640 EXPOSED WHILE THAT ONE COMPOUND 639 00:26:39,640 --> 00:26:41,400 IS BEING TRANSFERRED SO THIS 640 00:26:41,400 --> 00:26:43,400 SOLVES THAT PROBLEM AND THOSE 641 00:26:43,400 --> 00:26:46,120 COMPOUNDS WILL STAY FROZEN AND 642 00:26:46,120 --> 00:26:48,400 UNTOUCHED WHEN COMPOUND ORDER 643 00:26:48,400 --> 00:26:54,720 FULFILLMENT IS BEING DONE. 644 00:26:54,720 --> 00:26:59,120 YOU WILL HEAR US EXPLAIN THE 645 00:26:59,120 --> 00:26:59,440 WORKFLOWS. 646 00:26:59,440 --> 00:27:03,320 IN ARP IT'S THE COMPOUND IN 100% 647 00:27:03,320 --> 00:27:05,440 DMSO A SINGLE POINT OR ZERO 648 00:27:05,440 --> 00:27:06,920 DILUTION AND WHEN IT GETS INTO 649 00:27:06,920 --> 00:27:08,720 THE HANDS OF THE SCREENER THAT'S 650 00:27:08,720 --> 00:27:14,600 WHERE TH REAGENTS ARE ADD AND IT 651 00:27:14,600 --> 00:27:15,800 GOES INTO THE ASSAY. 652 00:27:15,800 --> 00:27:18,040 AN NARP IS FOR ANYTHING IT COULD 653 00:27:18,040 --> 00:27:21,080 BE SORT OF A PLATE THAT GETS 654 00:27:21,080 --> 00:27:22,200 TRANSFERRED INTO A DIFFERENT 655 00:27:22,200 --> 00:27:23,960 PLATE THAT IS THEN 656 00:27:23,960 --> 00:27:24,560 TRACTOR-TRAILER ASSAY READY 657 00:27:24,560 --> 00:27:28,040 FINAL PLAY OR MAYBE IT GETS THE 658 00:27:28,040 --> 00:27:29,360 BUFFER AND EVERYTHING ADDED 659 00:27:29,360 --> 00:27:31,200 BEFORE IT'S TRANSFERRED AND WITH 660 00:27:31,200 --> 00:27:35,800 THAT I'LL HAPPENED HAND IT OVER. 661 00:27:35,800 --> 00:27:37,840 >>IN OUR COMPOUND MANAGEMENT 662 00:27:37,840 --> 00:27:42,440 WORK FLOW WE START OUT IN A 663 00:27:42,440 --> 00:27:43,360 1.4-MILLILITER TUBE THAT IS 664 00:27:43,360 --> 00:27:46,720 STORED IN A HAMILTON VERSE OWE 665 00:27:46,720 --> 00:27:49,080 STORE IN COMPOUND MANAGEMENT 666 00:27:49,080 --> 00:27:50,640 ORGANIZATIONS THERE MAY BE 667 00:27:50,640 --> 00:27:52,200 DIFFERENT TYPES OF EQUIPMENT 668 00:27:52,200 --> 00:27:54,760 THAT HAVE THE ORGANIZATION FINDS 669 00:27:54,760 --> 00:27:56,880 FITS WELL UNDER THEIR WORK FLOW. 670 00:27:56,880 --> 00:27:59,040 WE FOUND THAT WE'VE BEEN ABLE TO 671 00:27:59,040 --> 00:28:00,320 STANDARDIZED WITH THE HAMILTON 672 00:28:00,320 --> 00:28:03,480 STAR AS THE TIP-BASED PLATFORM 673 00:28:03,480 --> 00:28:05,400 AND THE ECHO PRODUCTS FOR ACCUSE 674 00:28:05,400 --> 00:28:09,120 TICK DISPENSING INTO ASSAY READY 675 00:28:09,120 --> 00:28:12,520 PLATES. 676 00:28:12,520 --> 00:28:15,240 TYPICALLY FOR NARP PROCESS IT 677 00:28:15,240 --> 00:28:17,720 WOULD BE A STEP FOR ANY ASSAY 678 00:28:17,720 --> 00:28:18,560 READY PLATE PRODUCTION STEP, YOU 679 00:28:18,560 --> 00:28:20,240 CAN SEE THAT IT ALWAYS WOULD 680 00:28:20,240 --> 00:28:23,680 START WITH A TIP-BASED STEP ONE 681 00:28:23,680 --> 00:28:28,040 STARTING OUT FROM THE 1.4ML TO. 682 00:28:28,040 --> 00:28:32,320 WHEN WE MOVED TO THE ACOUSTIC 683 00:28:32,320 --> 00:28:34,440 TUBE IT WOULD REMOVE THE STEP 684 00:28:34,440 --> 00:28:38,040 FROM OUR ASSAY PRODUCTION 685 00:28:38,040 --> 00:28:38,440 WORKFLOWS. 686 00:28:38,440 --> 00:28:40,520 THERE'S ALWAYS A TIME WHEN USING 687 00:28:40,520 --> 00:28:44,880 A TIP-BASED TO DID A TRANSFER 688 00:28:44,880 --> 00:28:47,480 MAKES MORE SENSOR LESS THAN AN 689 00:28:47,480 --> 00:28:48,920 ACOUSTIC TRANSFER WE CALL THIS 690 00:28:48,920 --> 00:28:50,560 THE SOUND CHOICE. 691 00:28:50,560 --> 00:28:51,880 SO, ONE FACTS OR TO CONSIDER IS 692 00:28:51,880 --> 00:28:53,880 THE TIME IT TAKES TO DISPENSE SO 693 00:28:53,880 --> 00:28:56,840 WITH AN ECHO LIQUID HANDLER, 694 00:28:56,840 --> 00:29:00,480 DISPENSE NORTH 2.5 NANO LITER 695 00:29:00,480 --> 00:29:02,160 INCREMENTS THE HIGHER VOLUME YOU 696 00:29:02,160 --> 00:29:03,680 DISPENSE INCREASES THE TIME SO 697 00:29:03,680 --> 00:29:08,000 THE GRAPH ON THE TOP SHOWS THE 698 00:29:08,000 --> 00:29:12,480 DIFFERENCE IN TIME FOR ACCUSE 699 00:29:12,480 --> 00:29:15,320 TICK IN THE GREEN DOTS AND FOR 700 00:29:15,320 --> 00:29:19,880 TYPICAL BY PETTER THE TIME TO 701 00:29:19,880 --> 00:29:20,920 DISTANCE LIQUIDS IS CONSTANT. 702 00:29:20,920 --> 00:29:26,360 THE TARGET VOLUME ALSO EFFECTS 703 00:29:26,360 --> 00:29:29,200 THE CV THAT YOU CAN EXPERIENCE 704 00:29:29,200 --> 00:29:31,680 SO THE LOWER GRAPH SHOWS THAT 705 00:29:31,680 --> 00:29:36,840 WITH A TIP-BASED DIS SENSE YOU U 706 00:29:36,840 --> 00:29:38,600 CAN LOWER YOUR CV YOU WOULD SEE 707 00:29:38,600 --> 00:29:40,280 IN THOSE DISPENSES AT ABOUT ONE 708 00:29:40,280 --> 00:29:42,360 MIKE RE LITER YOU CAN SEE THOSE 709 00:29:42,360 --> 00:29:44,880 CROSS WHERE THE ACOUSTIC 710 00:29:44,880 --> 00:29:46,960 DISPENSER IS A CONSTANT AROUND 711 00:29:46,960 --> 00:29:51,400 3% WITH SOME VARIATIONS BUT THE 712 00:29:51,400 --> 00:29:53,400 TIP-BASED DISPENSER WILL DROP 713 00:29:53,400 --> 00:29:55,720 BELOW THAT IF YOU INCREASE THAT 714 00:29:55,720 --> 00:29:56,080 VOLUME. 715 00:29:56,080 --> 00:29:58,040 AND THE CONTAINERS AS WELL ARE 716 00:29:58,040 --> 00:30:00,560 LIMITATIONS SO YOU ARE ALWAYS 717 00:30:00,560 --> 00:30:03,800 REQUIRED TO USE ECHO QUALIFIED 718 00:30:03,800 --> 00:30:06,120 CONTAINERS WHICH HAVE SPECIFIC 719 00:30:06,120 --> 00:30:06,600 DEAD VOLUMES. 720 00:30:06,600 --> 00:30:08,680 SO, DEPENDING ON THE DENSITY OF 721 00:30:08,680 --> 00:30:10,280 YOUR CONTAINER, YOU CAN HAVE AS 722 00:30:10,280 --> 00:30:13,000 LOW AS A ONE MICRO LEADER DEAD 723 00:30:13,000 --> 00:30:15,000 VOLUME FOR A CONTAINER FOR AN 724 00:30:15,000 --> 00:30:17,240 ACOUSTIC DISPENSER BUT IT CAN 725 00:30:17,240 --> 00:30:19,680 INCREASE FROM THERE UP TO 15 726 00:30:19,680 --> 00:30:24,880 MICRO LITERS FOR THE ACOUSTIC 727 00:30:24,880 --> 00:30:25,240 TUBES. 728 00:30:25,240 --> 00:30:29,400 ONE THING ENABLED BY ACOUSTIC 729 00:30:29,400 --> 00:30:32,800 DISPENSING IS TYPICAL SERIAL DIE 730 00:30:32,800 --> 00:30:34,720 SOLUTION TRANSFERRING FROM ONE 731 00:30:34,720 --> 00:30:36,600 WELL OR CONTAINER TO THE NEXT 732 00:30:36,600 --> 00:30:39,840 AND A CONSTANT VOLUME WHEREAS 733 00:30:39,840 --> 00:30:42,120 DIRECTION DILUTION ALLOWS YOU TO 734 00:30:42,120 --> 00:30:43,280 DISTANCE THE ACTUAL AMOUNT OF 735 00:30:43,280 --> 00:30:45,800 VOLUME NEEDED TO ACHIEVE THE 736 00:30:45,800 --> 00:30:46,880 DESIRED CONCENTRATION IN A WELL 737 00:30:46,880 --> 00:30:49,880 AND USING A VARIABLE AMOUNT OF 738 00:30:49,880 --> 00:30:57,640 BUFFER TO BRING THAT UP TO THE 739 00:30:57,640 --> 00:30:58,360 DESIRED COMPOUND. 740 00:30:58,360 --> 00:31:04,600 THE BIGGEST IMPACT HERE IS WE 741 00:31:04,600 --> 00:31:08,640 DOUGH CRDECREASE THE ERROR. 742 00:31:08,640 --> 00:31:09,960 THERE ARE LIMITATIONS. 743 00:31:09,960 --> 00:31:11,840 THE TWO AND A HALF NANO LITER 744 00:31:11,840 --> 00:31:14,320 DROP SIZE DOES CAUSE A POTENTIAL 745 00:31:14,320 --> 00:31:18,240 DEVIATION FROM YOUR TARGET 746 00:31:18,240 --> 00:31:20,400 VOLUME, WHICH DOES CREATE A 747 00:31:20,400 --> 00:31:22,280 DEVIATION FROM YOUR 748 00:31:22,280 --> 00:31:25,280 CONCENTRATION OVER YOUR DILUTION 749 00:31:25,280 --> 00:31:25,480 RANGE. 750 00:31:25,480 --> 00:31:30,320 WE ARE ALSO LIMITED AND OUR 751 00:31:30,320 --> 00:31:31,320 DILUTION RANGE IN HOW LOW WE CAN 752 00:31:31,320 --> 00:31:34,120 ACHIEVE USING A SINGLE INTER 753 00:31:34,120 --> 00:31:34,840 MEDIATE PLATE. 754 00:31:34,840 --> 00:31:36,960 SOME PROCESSES MAY USE A SECOND 755 00:31:36,960 --> 00:31:39,120 PLATE BUT WE OPTED OUT OF THAT 756 00:31:39,120 --> 00:31:41,640 FOR SIMPLICITY IN OUR PROCESSES 757 00:31:41,640 --> 00:31:44,120 AND LIMIT THE LOWEST 758 00:31:44,120 --> 00:31:46,600 CONCENTRATION THAT WE CAN 759 00:31:46,600 --> 00:31:50,880 ACHIEVE FOR OUR DOWNSTREAM 760 00:31:50,880 --> 00:31:51,160 PROCESSES. 761 00:31:51,160 --> 00:31:53,640 DILUTION FACTOR IS SOMETHING TO 762 00:31:53,640 --> 00:31:55,080 CONSIDER, 3.162 IS A HALF LOG 763 00:31:55,080 --> 00:31:56,960 AND TYPICALLY IT'S SOMETHING 764 00:31:56,960 --> 00:31:59,920 THAT IS USED FREQUENTLY IN 765 00:31:59,920 --> 00:32:02,320 SCREENING BUT WE FOUND ACTUALLY 766 00:32:02,320 --> 00:32:04,360 USING WHOLE NUMBERS DUE TO THE 767 00:32:04,360 --> 00:32:06,320 2.5 NANO LITER DROP SIZE IS MORE 768 00:32:06,320 --> 00:32:10,600 ADD VADVANTAGEOUS. 769 00:32:10,600 --> 00:32:12,760 THE VOLUME HERE IS A FACTOR AS 770 00:32:12,760 --> 00:32:15,240 WELL SO THE LARGER VOLUMES DO 771 00:32:15,240 --> 00:32:18,720 TAKE LONGER TO TRANSFER. 772 00:32:18,720 --> 00:32:20,400 SO IT'S SOMETHING TO CONSIDER 773 00:32:20,400 --> 00:32:22,440 WHEN WE WORK WITH OUR DOWN 774 00:32:22,440 --> 00:32:25,280 STREAM SCREENERS WHEN SETTING UP 775 00:32:25,280 --> 00:32:27,880 THEIR SUBMITALS IS THE NUMBER OF 776 00:32:27,880 --> 00:32:29,000 DILUTION POINTS AND DILUTION 777 00:32:29,000 --> 00:32:30,880 FACTORS SO AS I MENTIONED, YOU 778 00:32:30,880 --> 00:32:32,280 CAN SEE THAT THERE'S A 779 00:32:32,280 --> 00:32:34,920 DIFFERENCE HERE IN THE WHOLE 780 00:32:34,920 --> 00:32:36,600 NUMBERS ON THE LEFT AND RIGHT 781 00:32:36,600 --> 00:32:38,360 AND THE HALF LOG DILUTION YOU 782 00:32:38,360 --> 00:32:41,720 CAN SEE A MUCH BOUNCIER 783 00:32:41,720 --> 00:32:44,000 DEVIATION FROM THE TARGET 784 00:32:44,000 --> 00:32:44,480 CONCENTRATION. 785 00:32:44,480 --> 00:32:46,280 SO YOUR TAKE HOMES HERE ARE 786 00:32:46,280 --> 00:32:49,160 REALLY THAT ONE THING IS THAT A 787 00:32:49,160 --> 00:32:50,360 10-POINT DILUTION IS BETTER THAN 788 00:32:50,360 --> 00:32:53,120 11, ESPECIALLY IF YOU ARE AT A 789 00:32:53,120 --> 00:32:54,520 HIGHER DILUTION FACTOR AND YOUR 790 00:32:54,520 --> 00:32:58,040 WHOLE NUMBER GIVES YOU GENERALLY 791 00:32:58,040 --> 00:33:01,400 A CLOSER TO YOUR TARGET. 792 00:33:01,400 --> 00:33:04,360 WHEN YOU ARE LOOKING AT YOUR TOP 793 00:33:04,360 --> 00:33:06,080 CONCENTRATION AND THE VOLUME OF 794 00:33:06,080 --> 00:33:07,240 YOUR DELIVERABLE, THESE THINGS 795 00:33:07,240 --> 00:33:09,560 GO HAND AND HAND AS WELL AND 796 00:33:09,560 --> 00:33:12,080 WHETHER YOU ARE WORKING WITH A 797 00:33:12,080 --> 00:33:14,080 FIXED CONCENTRATION INVENTORY OR 798 00:33:14,080 --> 00:33:16,000 YOU ARE ABLE TO DILUTE YOUR 799 00:33:16,000 --> 00:33:18,040 SOURCE TO THE TOP CONCENTRATION, 800 00:33:18,040 --> 00:33:20,200 YOU CAN ALSO EFFECT YOUR ABILITY 801 00:33:20,200 --> 00:33:25,960 TO HIT THOSE LE LOWER 802 00:33:25,960 --> 00:33:30,440 CONCENTRATIONS IN YOUR DILUTION. 803 00:33:30,440 --> 00:33:34,480 YOU CAN INCREASE YOUR VOLUME TO 804 00:33:34,480 --> 00:33:35,840 ACCOMMODATE THAT AND TO GET TO 805 00:33:35,840 --> 00:33:38,680 THOSE LOWER CONCENTRATIONS AND 806 00:33:38,680 --> 00:33:41,880 FOR A THREEFOLD DILUTION, THE 807 00:33:41,880 --> 00:33:45,080 MULTIPLE OF 22.5 NANO LITER IS 808 00:33:45,080 --> 00:33:48,320 BEST AND 40 AND I DERIVED THAT 809 00:33:48,320 --> 00:33:50,160 FROM THE GUIDELINE CALCULATION 810 00:33:50,160 --> 00:33:56,200 UP AT THE TOP WHICH DICK TAKES A 811 00:33:56,200 --> 00:33:58,040 GOOD TARTING PLACE THAT A ASSAY 812 00:33:58,040 --> 00:34:01,440 READY PLATE MIGHT BE READY TO 813 00:34:01,440 --> 00:34:01,840 ACHIEVE. 814 00:34:01,840 --> 00:34:09,040 IT'S VERY IMPORTANT FOR US TO 815 00:34:09,040 --> 00:34:17,520 MAINTAIN WE USING SINGLE SHELF 816 00:34:17,520 --> 00:34:21,960 AND WE HAVE A HOME-BUILT SERIAL 817 00:34:21,960 --> 00:34:27,600 DILUTION THAT WE DO THAT IS 818 00:34:27,600 --> 00:34:29,280 EXCEL BASED AND VALUES AND CAN 819 00:34:29,280 --> 00:34:31,200 GIVE US THE HEALTH VALUE IN HOW 820 00:34:31,200 --> 00:34:34,040 WELL OUR SYSTEMS ARE MAKING 821 00:34:34,040 --> 00:34:37,240 SERIAL AND DIRECT DILUTION 822 00:34:37,240 --> 00:34:37,720 DELIVERABLES. 823 00:34:37,720 --> 00:34:40,080 YOU CAN SEE HERE, IN THIS GRAPH 824 00:34:40,080 --> 00:34:42,160 AND EXAMPLE OF WHERE WE 825 00:34:42,160 --> 00:34:44,160 IDENTIFIED A PROBLEM, SO YOU CAN 826 00:34:44,160 --> 00:34:46,280 SEE OVER TIME, WE'RE ABLE TO 827 00:34:46,280 --> 00:34:49,120 LOOK AT QC VALUES FROM OUR ECHOS 828 00:34:49,120 --> 00:34:53,120 AND YOU CAN SEE ON THE TOP, A 829 00:34:53,120 --> 00:34:54,800 NICE TREND OVER TIME WHERE WE'RE 830 00:34:54,800 --> 00:34:56,560 NOT SEEING A LOT OF VARIATION 831 00:34:56,560 --> 00:34:58,560 AND ON THE BOTTOM, AN EXAMPLE 832 00:34:58,560 --> 00:35:09,080 FROM A HAMILTON WHERE AND YOU 833 00:35:13,920 --> 00:35:16,880 CAN SEE THE VALUES BACK TO A 834 00:35:16,880 --> 00:35:17,960 HISTORICAL NORMAL VALUE AFTER 835 00:35:17,960 --> 00:35:19,880 THAT SERVICE WAS CREED. 836 00:35:19,880 --> 00:35:21,680 WITHIN COMPOUND MANAGEMENT, WE 837 00:35:21,680 --> 00:35:24,320 TRACK ALL OF OUR PROCESSES AND 838 00:35:24,320 --> 00:35:26,320 USING ONE AND TWO D BAR CODES 839 00:35:26,320 --> 00:35:28,200 THROUGH OUT EVERY STEP OF THE 840 00:35:28,200 --> 00:35:30,640 WAY AND WE USE LABORATORY 841 00:35:30,640 --> 00:35:34,200 INFORMATION MANAGEMENT SYSTEMS 842 00:35:34,200 --> 00:35:35,960 TO TRACK THOSE IN EVERY STEP OF 843 00:35:35,960 --> 00:35:38,280 THE PROCESS AND YOU CAN SEE AN 844 00:35:38,280 --> 00:35:40,280 EXAMPLE HERE OF A USER INTERFACE 845 00:35:40,280 --> 00:35:44,720 AND ONE OF OUR SYSTEMS THAT 846 00:35:44,720 --> 00:35:46,800 ALLOWS US TO ESSENTIALLY TURN 847 00:35:46,800 --> 00:35:49,080 EVERY STEP OF THE OF OUR PROCESS 848 00:35:49,080 --> 00:35:50,800 INTO A BUSINESSARD THAT CAN BE 849 00:35:50,800 --> 00:35:52,240 GIVEN BY ANY MEMBER OF THE LAB 850 00:35:52,240 --> 00:35:53,920 AND ONE THING WE FOUND WITH THIS 851 00:35:53,920 --> 00:35:55,920 IS REALLY REDUCES OUR TRAINING 852 00:35:55,920 --> 00:35:58,680 BURDEN AND WHEN WE HAVE STAFF 853 00:35:58,680 --> 00:36:00,800 TURNOVER AND WE'RE ABLE TO 854 00:36:00,800 --> 00:36:02,960 QUICKLY BRING OUR STAFF UP TO 855 00:36:02,960 --> 00:36:06,040 SPEED AND MAKE THEM ABLE TO 856 00:36:06,040 --> 00:36:16,200 PROCESS OUR DELIVERABLES. 857 00:36:16,200 --> 00:36:17,840 WE HAVE PROCESS ANOMALY THAT'S 858 00:36:17,840 --> 00:36:21,920 WE HAVE TO TROUBLE SHOOT SO 859 00:36:21,920 --> 00:36:25,320 OCCASIONALLY, A DOWNSTREAM 860 00:36:25,320 --> 00:36:26,560 CUSTOMER COMES AND SAYS THEY 861 00:36:26,560 --> 00:36:27,920 FOUND SOMETHING OUT OF ORDINARY 862 00:36:27,920 --> 00:36:33,680 AND WE USED TOOLS WITHIN OUR 863 00:36:33,680 --> 00:36:37,920 TOOLBOX TO ANALYZE THE PROCESS 864 00:36:37,920 --> 00:36:40,600 AND DETERMINE WHERE IT OCCURRED. 865 00:36:40,600 --> 00:36:43,480 ONE GREAT EXAMPLE IS THE ECHO 866 00:36:43,480 --> 00:36:45,880 TRANSFER LOB AND WE'LL USE OUR 867 00:36:45,880 --> 00:36:50,000 LIMB SYSTEM TO DETERMINE WHICH 868 00:36:50,000 --> 00:36:51,480 ECHO THE PROCESS OCCURRED ON AND 869 00:36:51,480 --> 00:36:54,040 WE CAN THEN GO LOCK AT THAT LOG 870 00:36:54,040 --> 00:36:59,640 TO POTENTIALLY DETERMINE A 871 00:36:59,640 --> 00:36:59,880 FAILURE. 872 00:36:59,880 --> 00:37:03,640 WE CAN ALSO CHECK QC AND 873 00:37:03,640 --> 00:37:04,840 DETERMINE THE PERFORMANCE OF 874 00:37:04,840 --> 00:37:06,680 THAT INSTRUMENT AT THAT TIME AS 875 00:37:06,680 --> 00:37:07,240 WELL. 876 00:37:07,240 --> 00:37:09,600 SO, JUST TO WRAP THINGS UP, 877 00:37:09,600 --> 00:37:11,200 SAMPLE STORAGE IS SOMETHING THAT 878 00:37:11,200 --> 00:37:12,960 IS VERY IMPORTANT AND IN 879 00:37:12,960 --> 00:37:16,160 COMPOUND MANAGEMENT AND TO 880 00:37:16,160 --> 00:37:17,960 PROLONG THE LIFE OF THE 881 00:37:17,960 --> 00:37:19,920 COLLECTION AND AS LONG AS 882 00:37:19,920 --> 00:37:22,640 POSSIBLE AND IN ORDER TO 883 00:37:22,640 --> 00:37:23,600 DETERMINE HOW LONG OUR 884 00:37:23,600 --> 00:37:26,000 COLLECTION CAN SURVIVE, SAMPLE 885 00:37:26,000 --> 00:37:28,960 QC IS SOMETHING THAT WE HAVE TO 886 00:37:28,960 --> 00:37:32,800 DO REGULARLY AS WELL AS 887 00:37:32,800 --> 00:37:34,920 MAINTAINING AND QCs OUR 888 00:37:34,920 --> 00:37:36,600 INNSTATION TO ENSURE 889 00:37:36,600 --> 00:37:38,520 "HIGH-QUALITY DELIVERABLES AND 890 00:37:38,520 --> 00:37:42,960 THE DATA HANDLING ALONG THE 891 00:37:42,960 --> 00:37:44,920 WHOLE PROCESS IN ORDER TO TRACK 892 00:37:44,920 --> 00:37:49,720 THE COMPOUNDS ALONG THE WAY. 893 00:37:49,720 --> 00:37:52,320 SO, I'D LIKE TO THANK SOME OTHER 894 00:37:52,320 --> 00:37:54,080 MEMBERS OF OUR TEAM, OUR 895 00:37:54,080 --> 00:37:55,440 COMPOUND MANAGEMENT AND 896 00:37:55,440 --> 00:37:56,440 DISTRIBUTION TEAM. 897 00:37:56,440 --> 00:37:58,320 I WON'T LIST ALL OF THEM BUT 898 00:37:58,320 --> 00:38:01,840 EVERYONE HAS CONTRIBUTED TO OUR 899 00:38:01,840 --> 00:38:03,400 EXPERTISE HERE AND OUR 900 00:38:03,400 --> 00:38:05,720 ADDITIONAL COLLABORATORS, ROSE, 901 00:38:05,720 --> 00:38:07,880 THE FORMER DIRECTOR OF OUR GROUP 902 00:38:07,880 --> 00:38:16,160 AND HOLLY McKEE, WHO WAS. 903 00:38:16,160 --> 00:38:17,960 SO WITH THAT WE'D LIKE TO INVITE 904 00:38:17,960 --> 00:38:21,160 QUESTIONS THAT YOU MIGHT HAVE. 905 00:38:21,160 --> 00:38:23,200 >>THANK YOU, VERY MUCH. 906 00:38:23,200 --> 00:38:25,520 TRAVIS AND LANE. 907 00:38:25,520 --> 00:38:31,840 [APPLAUSE] 908 00:38:31,840 --> 00:38:32,640 WE HAVE QUESTIONS. 909 00:38:32,640 --> 00:38:34,160 I'M GOING TO TAKE THE MICROPHONE 910 00:38:34,160 --> 00:38:40,360 TO THE FIRST QUESTION. 911 00:38:40,360 --> 00:38:42,520 >>THANK YOU FOR THE NICE 912 00:38:42,520 --> 00:38:43,000 PRESENTATION. 913 00:38:43,000 --> 00:38:45,840 I'M JUST CURIOUS, DURING 914 00:38:45,840 --> 00:38:47,720 AUTOMATIC HANDLING OF THE 915 00:38:47,720 --> 00:38:49,720 COMPOUND, HOW DOES IT HANDLE THE 916 00:38:49,720 --> 00:38:54,880 STICKY COMPOUND TO TRANSFER THE 917 00:38:54,880 --> 00:38:56,600 EXACT AMOUNT AND ALSO IT'S 918 00:38:56,600 --> 00:38:58,720 DIFFICULT TO HANDLE THE VERY 919 00:38:58,720 --> 00:39:00,720 STICKY COMPOUND AND ANOTHER 920 00:39:00,720 --> 00:39:03,800 THING WHEN WE HAVE DIFFERENT 921 00:39:03,800 --> 00:39:05,240 COMPOUNDS, SAME COMPOUND DO WE 922 00:39:05,240 --> 00:39:08,200 KEEP THAT COMPOUND IN THE SAME 923 00:39:08,200 --> 00:39:11,280 PART OR IT'S A DIFFERENT BECAUSE 924 00:39:11,280 --> 00:39:16,400 IT COULD BE LOOKS THE SAME. 925 00:39:16,400 --> 00:39:21,400 THANK YOU. 926 00:39:21,400 --> 00:39:23,160 >>THIS IS A TRICKY ONE. 927 00:39:23,160 --> 00:39:25,360 I THINK THERE'S MANY SCHOOLS OF 928 00:39:25,360 --> 00:39:28,120 THOUGHT ON THAT AND OURS IS 929 00:39:28,120 --> 00:39:31,000 TYPICALLY THAT IF IT'S TOO 930 00:39:31,000 --> 00:39:33,480 STICKY TO DISSOLVE IT ALL IN 931 00:39:33,480 --> 00:39:35,320 DMSO THAT THAT IS SOMETHING THAT 932 00:39:35,320 --> 00:39:37,720 MIGHT BE CAUGHT IN OUR QC 933 00:39:37,720 --> 00:39:40,680 PROCESS AND THAT IS FLAGGED. 934 00:39:40,680 --> 00:39:43,480 TYPICALLY, I THINK WE FIND THAT 935 00:39:43,480 --> 00:39:45,840 THOSE COMPOUNDS JUST EXPERIENCE 936 00:39:45,840 --> 00:39:49,640 A LOSS OF CONCENTRATION AND SO 937 00:39:49,640 --> 00:39:51,800 WORE ABLE TO PROCESS THOSE 938 00:39:51,800 --> 00:39:55,320 THROUGH MOSTLY THROUGH OUR 939 00:39:55,320 --> 00:39:58,360 NORMAL PROCESSES AND THROUGH 940 00:39:58,360 --> 00:39:59,920 DOWNSTREAM ANALYSIS DETERMINED 941 00:39:59,920 --> 00:40:03,720 WHAT WE NEED TO. 942 00:40:03,720 --> 00:40:05,880 >>SO, AGAIN, TO SORT OF RELATES 943 00:40:05,880 --> 00:40:07,560 TO THE STICKY COMPOUNDS AS WELL 944 00:40:07,560 --> 00:40:08,040 RIGHT. 945 00:40:08,040 --> 00:40:11,120 SO IF YOU ARE EXPECTING SOME 946 00:40:11,120 --> 00:40:12,760 SORT OF VALUE IN YOUR ASSAY FROM 947 00:40:12,760 --> 00:40:14,200 THAT COMPOUND AND YOU ARE 948 00:40:14,200 --> 00:40:16,280 GETTING IT, THAT'S WHERE WE 949 00:40:16,280 --> 00:40:17,240 WOULD BACKTRACK THROUGH OUR 950 00:40:17,240 --> 00:40:17,480 PROCESS. 951 00:40:17,480 --> 00:40:20,080 A LOT OF TIMES I THINK, THE ECHO 952 00:40:20,080 --> 00:40:21,720 WOULD IT TRANSFER OR NOT 953 00:40:21,720 --> 00:40:22,520 TRANSFER? 954 00:40:22,520 --> 00:40:23,200 TYPICALLY IT WOULD BE -- 955 00:40:23,200 --> 00:40:28,600 >>THE ECHO WE MAY SEE AN 956 00:40:28,600 --> 00:40:29,560 INFRACTION AND WE'RE ABLE TO 957 00:40:29,560 --> 00:40:31,200 FLAG IT IF THE ECHO WILL 958 00:40:31,200 --> 00:40:32,960 DETERMINE THAT THERE'S SOME 959 00:40:32,960 --> 00:40:34,800 ANOMALY WITH THE LIQUID AND 960 00:40:34,800 --> 00:40:36,200 UNABLE TO DISPENSE IT. 961 00:40:36,200 --> 00:40:41,400 ONE THING OUR LIMP SYSTEM IS IT 962 00:40:41,400 --> 00:40:43,200 INTEG GATES THE PROFILE FROM THE 963 00:40:43,200 --> 00:40:45,000 ECHO AND THE ECHO REPORTS THAT 964 00:40:45,000 --> 00:40:47,280 IT WAS UNABLE TO MAKE A TRANSFER 965 00:40:47,280 --> 00:40:49,080 FOR PARTICULAR COMPOUNDS, WE DO 966 00:40:49,080 --> 00:40:51,080 CAPTURE THAT AND WE PUBLISH THAT 967 00:40:51,080 --> 00:40:53,600 BACK TO THE SYSTEMS SO THAT THE 968 00:40:53,600 --> 00:40:54,760 RECIPIENT OF THOSE COMPOUNDS 969 00:40:54,760 --> 00:40:56,920 KNOWS THAT THAT COMPOUND IS NOT 970 00:40:56,920 --> 00:40:58,120 ACTUALLY PRESENT IN THAT WELL 971 00:40:58,120 --> 00:40:59,520 WHERE THEY EXPECTED IT AND WE 972 00:40:59,520 --> 00:41:02,560 HAVE DIFFERENT WAYS OF HANDLING 973 00:41:02,560 --> 00:41:02,760 IT. 974 00:41:02,760 --> 00:41:05,080 AS FAR AS BATCH TO BATCH 975 00:41:05,080 --> 00:41:08,360 VARIATION, WE HAVE DIFFERENT 976 00:41:08,360 --> 00:41:11,880 WAYS OF HANDLING IT, JUST TO 977 00:41:11,880 --> 00:41:13,200 REFLECT BACK TO THAT OTHER SIDE 978 00:41:13,200 --> 00:41:17,600 THAT LANE SHOWED YOU WITH THE 979 00:41:17,600 --> 00:41:19,960 21.4 TUBE, THE UPSTREAM 980 00:41:19,960 --> 00:41:21,040 PURIFICATION PROCESS HAS 981 00:41:21,040 --> 00:41:22,280 BASICALLY STANDARDIZED ON THAT 982 00:41:22,280 --> 00:41:24,920 CONTAINER SO ALL OF OUR NEW 983 00:41:24,920 --> 00:41:26,440 COMPOUNDS COME IN AND THAT 984 00:41:26,440 --> 00:41:30,120 CONTAINER AND IF A SYNTHESIZED 985 00:41:30,120 --> 00:41:32,720 COMPOUND HAS TWO DIFFERENT 986 00:41:32,720 --> 00:41:34,120 BATCHES IN THOSE TUBES IN THAT 987 00:41:34,120 --> 00:41:37,080 SYSTEM WE CAN RUN ADDITIONAL 988 00:41:37,080 --> 00:41:38,760 ANALYSIS AND THEY SAY THIS IS A 989 00:41:38,760 --> 00:41:40,360 PROBLEM, AND THEY CAN ACTUALLY 990 00:41:40,360 --> 00:41:42,000 REQUEST TO FULL AND WE CAN PULL 991 00:41:42,000 --> 00:41:43,680 THAT TUBE AND RETURN IT TO THEM 992 00:41:43,680 --> 00:41:45,800 AND WE CAN THROW IT OUT SO WE DO 993 00:41:45,800 --> 00:41:46,840 HAVE A PROCESS IN PLACE FOR 994 00:41:46,840 --> 00:41:48,040 THAT, HOWEVER, WHEN THOSE 995 00:41:48,040 --> 00:41:49,440 COMPOUNDS HAVE GONE INTO OUR 996 00:41:49,440 --> 00:41:50,760 LONG-TERM STORAGE, THEY'RE NOW 997 00:41:50,760 --> 00:41:52,280 SPREAD ACROSS MANY CONTAINERS 998 00:41:52,280 --> 00:41:53,640 AND WE'VE HAD THIS ISSUE AS 999 00:41:53,640 --> 00:41:53,920 WELL. 1000 00:41:53,920 --> 00:41:58,400 WE'VE HAD A CASE WHERE CHEMIST 1001 00:41:58,400 --> 00:41:59,960 SAY THIS IS NOT GOOD CAN YOU 1002 00:41:59,960 --> 00:42:00,640 REMOVE IT? 1003 00:42:00,640 --> 00:42:02,080 WE CAN'T PHYSICALLY REMOVE IT 1004 00:42:02,080 --> 00:42:04,200 BECAUSE IT'S SPREAD ACROSS A 1005 00:42:04,200 --> 00:42:05,960 THOUSAND CONTAINERS, I'M 1006 00:42:05,960 --> 00:42:07,640 EXAGGERATING BUT WE HAVE A 1007 00:42:07,640 --> 00:42:09,560 MECHANISM TO DELETE IT 1008 00:42:09,560 --> 00:42:10,240 ELECTRONICALLY. 1009 00:42:10,240 --> 00:42:11,760 AS WE RECOMMEND TO OUR 1010 00:42:11,760 --> 00:42:13,560 SCREENERS, ALWAYS CHECK THE QC 1011 00:42:13,560 --> 00:42:14,120 DATA. 1012 00:42:14,120 --> 00:42:15,680 IT COULD BE THAT IT'S NOT 1013 00:42:15,680 --> 00:42:17,000 ELECTRONICALLY DELETED THAT 1014 00:42:17,000 --> 00:42:18,280 MATCH IS IN OUR FILE, THEY 1015 00:42:18,280 --> 00:42:20,640 RECEIVED IT AND THE QC DATA SAYS 1016 00:42:20,640 --> 00:42:21,720 FAIL, FAIL, FAIL. 1017 00:42:21,720 --> 00:42:23,520 ONE THING TO NOTE ALSO WITH THE 1018 00:42:23,520 --> 00:42:25,440 PURITY, WE USE THE PURITY CUT 1019 00:42:25,440 --> 00:42:27,200 OFF OF 80%. 1020 00:42:27,200 --> 00:42:28,480 THERE'S BEEN OTHER SIDE STUDIES 1021 00:42:28,480 --> 00:42:32,840 THAT HAVE SHOWN THAT BIOCHEMICAL 1022 00:42:32,840 --> 00:42:35,280 ASSAYS CAN GET RELY ABLE RESULTS 1023 00:42:35,280 --> 00:42:36,760 RATHER THE CELL BASED ASSAYS ARE 1024 00:42:36,760 --> 00:42:39,560 A LOT MORE SENSITIVE TO THAT AND 1025 00:42:39,560 --> 00:42:42,280 ALTERNATIVELY, OUR MASS SPECK 1026 00:42:42,280 --> 00:42:45,800 END POINT CAN TOLERATE A 50% 1027 00:42:45,800 --> 00:42:47,520 PURE COMP FOUND SO IT'S NOT JUST 1028 00:42:47,520 --> 00:42:48,960 READING THE PASS-FAIL SUSPECT 1029 00:42:48,960 --> 00:42:52,160 BUT GET THE REAL DATA AND KNOW 1030 00:42:52,160 --> 00:42:54,160 WHAT THAT PURIFICATION IS OF 1031 00:42:54,160 --> 00:42:55,400 THAT COMPOUND SO THERE'S A LOT 1032 00:42:55,400 --> 00:42:57,800 OF DIGGING YOU NEED TO DO BUT WE 1033 00:42:57,800 --> 00:42:59,400 HAVE A MECHANISM FOR IF THERE'S 1034 00:42:59,400 --> 00:43:01,080 DEFINITELY A BAD BATCH, WE HAVE 1035 00:43:01,080 --> 00:43:02,920 A WAY OF EITHER PHYSICALLY 1036 00:43:02,920 --> 00:43:04,560 REMOVING IT OR ELECTRONICALLY 1037 00:43:04,560 --> 00:43:08,880 REMOVING IT FROM OUR COLLECTION. 1038 00:43:08,880 --> 00:43:10,480 >>TWO QUICK QUESTIONS, ONE IS 1039 00:43:10,480 --> 00:43:12,520 WITH REGARD TO YOUR WEIGHING 1040 00:43:12,520 --> 00:43:17,800 GREATER THAN TWO MIGS, WHAT IS 1041 00:43:17,800 --> 00:43:21,800 THE NUMBER YOU WEIGH TOO? 1042 00:43:21,800 --> 00:43:23,720 WE DON'T HANDLE DRIVES IN OUR 1043 00:43:23,720 --> 00:43:25,600 WORK FLOW, WE ACTUALLY ONLY, 1044 00:43:25,600 --> 00:43:27,000 WELL WE DO PHYSICALLY HAVE A 1045 00:43:27,000 --> 00:43:29,560 BALANCE BUT WE TELL PEOPLE WE 1046 00:43:29,560 --> 00:43:29,880 DON'T. 1047 00:43:29,880 --> 00:43:33,600 ALL OF OUR STAFF IS BASICALLY 1048 00:43:33,600 --> 00:43:35,840 OUTSOURCES TO SIGMA AND THEY 1049 00:43:35,840 --> 00:43:37,040 HAVE, THEY DO HAVE BALANCE 1050 00:43:37,040 --> 00:43:41,280 THAT'S CAN GO DOWN TO ONE MIG 1051 00:43:41,280 --> 00:43:43,080 AND LOWER BUT WE'VE EXPERIENCED 1052 00:43:43,080 --> 00:43:45,080 ISSUES ON THAT AND THAT IS WE 1053 00:43:45,080 --> 00:43:48,720 STANDARDIZED ON THE TWO PIGS -- 1054 00:43:48,720 --> 00:43:54,600 >>IS IT 1.25MIGS OR? 1055 00:43:54,600 --> 00:44:02,040 >>IT'S TWO. 1056 00:44:02,040 --> 00:44:04,400 SO DUE WEIGH POWDER SO YOU CAN 1057 00:44:04,400 --> 00:44:06,080 WORK WITH OIL AS LONG AS YOU 1058 00:44:06,080 --> 00:44:07,280 KNOW THE WEIGHT? 1059 00:44:07,280 --> 00:44:09,920 >>WE DON'T REWEIGH. 1060 00:44:09,920 --> 00:44:13,160 >>CORRECT. 1061 00:44:13,160 --> 00:44:19,080 >>HAVE YOU RIDE TAKING THE 1062 00:44:19,080 --> 00:44:22,520 COMPOUNDS OUT AT ROOM 1063 00:44:22,520 --> 00:44:25,480 TEMPERATURE AND DO YOU SEE A 1064 00:44:25,480 --> 00:44:25,840 PROBLEM? 1065 00:44:25,840 --> 00:44:27,720 >>WE DIDN'T PUT IT IN HERE. 1066 00:44:27,720 --> 00:44:29,560 WE HAD THEM AT ROM TEMPERATURE 1067 00:44:29,560 --> 00:44:33,200 FOR TWO YEARS AND IT WAS AT 1068 00:44:33,200 --> 00:44:34,840 WHERE THE REAL PROBLEMS REALLY 1069 00:44:34,840 --> 00:44:36,640 STARTED TO SHOW UP MOST 1070 00:44:36,640 --> 00:44:38,520 SIGNIFICANTLY WAS 18 MONTHS AND 1071 00:44:38,520 --> 00:44:40,280 OUR RECOMMENDATION WAS TO 1072 00:44:40,280 --> 00:44:41,880 BASICALLY RE FRESH THEM AFTER A 1073 00:44:41,880 --> 00:44:43,520 YEAR SO OUR CURRENT SAMPLES THAT 1074 00:44:43,520 --> 00:44:45,240 WE HAVE SO WE DON'T GET INTO 1075 00:44:45,240 --> 00:44:46,680 THAT TROUBLE ZONE BUT WHEN WE 1076 00:44:46,680 --> 00:44:48,200 HAVE OUR SAMPLES AT ROOM 1077 00:44:48,200 --> 00:44:49,520 TEMPERATURE ON OUR CURRENT 1078 00:44:49,520 --> 00:44:51,440 SYSTEM, WE REFRESH THEM ONCE A 1079 00:44:51,440 --> 00:44:53,120 YEAR SO, THAT'S WHERE WE'RE AT 1080 00:44:53,120 --> 00:44:53,280 NOW. 1081 00:44:53,280 --> 00:44:54,640 >>THERE'S NO TWIST. 1082 00:44:54,640 --> 00:44:56,960 >>YEAH. 1083 00:44:56,960 --> 00:44:58,960 >>IT'S DEGRADATION. 1084 00:44:58,960 --> 00:45:01,040 >>AT ABOUT 18 MONTHS IT STARTS 1085 00:45:01,040 --> 00:45:02,440 TO BE MUCH MORE NOTICEABLE AND 1086 00:45:02,440 --> 00:45:06,440 AT TWO YEARS, WE ACTUALLY DID 1087 00:45:06,440 --> 00:45:11,160 ONE STUDY UP TO THROW YE THREE T 1088 00:45:11,160 --> 00:45:13,720 IT WAS WORST-CASE SCENARIO. 1089 00:45:13,720 --> 00:45:15,160 OUR RECOMMENDATION IS USE A YEAR 1090 00:45:15,160 --> 00:45:17,080 IF YOU HAVE TO PUSH IT OUT THEN 1091 00:45:17,080 --> 00:45:19,800 DON'T GO PASSED 18 MONTHS. 1092 00:45:19,800 --> 00:45:23,360 >>AND WHAT DO YOU THINK ABOUT 1093 00:45:23,360 --> 00:45:25,480 JUST SATURATING YOUR DMSO TO 1094 00:45:25,480 --> 00:45:27,600 BEGIN WITH SINCE IT SAT RATES SO 1095 00:45:27,600 --> 00:45:28,960 QUICKLY FOR EXPOSURE. 1096 00:45:28,960 --> 00:45:33,080 WHAT DO YOU THINK ABOUT THAT? 1097 00:45:33,080 --> 00:45:34,160 >>SO -- 1098 00:45:34,160 --> 00:45:34,600 [LAUGHTER] 1099 00:45:34,600 --> 00:45:37,200 IN ONE OF MY PAST POSITIONS AT 1100 00:45:37,200 --> 00:45:40,800 PFIZER, WE DID DO THAT. 1101 00:45:40,800 --> 00:45:42,680 FOR THAT WRACK FLOW, IT DID WORK 1102 00:45:42,680 --> 00:45:43,840 PRETTY WELL. 1103 00:45:43,840 --> 00:45:46,840 THE THING THERE IS YOU DO HAVE 1104 00:45:46,840 --> 00:45:48,600 THE DIFFERENT KINDS OF -- THE 1105 00:45:48,600 --> 00:45:51,080 WAY THAT DMSO PERFORMS SO YOU 1106 00:45:51,080 --> 00:45:53,920 WON'T FREEZE IT SO EQUILIBRIUM 1107 00:45:53,920 --> 00:45:57,120 EVEN AT NEGATIVE 140C YOU WILL 1108 00:45:57,120 --> 00:45:57,600 GET A GEL. 1109 00:45:57,600 --> 00:45:59,920 WE DON'T HAVE A LOT OF DATA ON 1110 00:45:59,920 --> 00:46:00,400 THAT. 1111 00:46:00,400 --> 00:46:01,920 ONE THING WE USED TO DO IS WE 1112 00:46:01,920 --> 00:46:07,520 USED TO ADD AN YOU HAVE TO PLAY 1113 00:46:07,520 --> 00:46:09,160 WITH IT AND BASED ON THE 1114 00:46:09,160 --> 00:46:11,280 RECOMMENDATION AND STUDY AND HOW 1115 00:46:11,280 --> 00:46:13,200 WE'VE AND I DON'T KNOW IF YOU'VE 1116 00:46:13,200 --> 00:46:15,000 SEEN CHRIS LA PIN SKI'S FUEL AND 1117 00:46:15,000 --> 00:46:17,720 ALL THE OTHER THINGS BUT PULLING 1118 00:46:17,720 --> 00:46:18,800 THINGS TOGETHER IT WAS TO KEEP 1119 00:46:18,800 --> 00:46:22,000 IT IN 100% DMSO FOR AS LONG AS 1120 00:46:22,000 --> 00:46:23,160 POSSIBLE AND THE KEEP THE WATER 1121 00:46:23,160 --> 00:46:23,920 OUT OF IT. 1122 00:46:23,920 --> 00:46:27,760 >>ONE THING I WOULD MENTION 1123 00:46:27,760 --> 00:46:29,200 ABOUT THAT IN PARTICULAR IS THE 1124 00:46:29,200 --> 00:46:32,640 ECHO LIMIT OF DETECTION IS 70% 1125 00:46:32,640 --> 00:46:35,600 AND SO IF YOU ARE AT EQUILIBRIUM 1126 00:46:35,600 --> 00:46:39,640 YOU WILL LIKELY EXPERIENCE MANY 1127 00:46:39,640 --> 00:46:42,600 MORE FAILURES AND UTILIZING 1128 00:46:42,600 --> 00:46:42,880 ACOUSTICS. 1129 00:46:42,880 --> 00:46:45,000 >>IN THE PAST WHEN IT WAS ALL 1130 00:46:45,000 --> 00:46:46,600 TIP BASED THAT WASN'T SUCH A BIG 1131 00:46:46,600 --> 00:46:49,840 CONCERN AND I THINK IT WAS NICE 1132 00:46:49,840 --> 00:46:51,040 TO HAVE EVERYTHING UNIFORM BUT 1133 00:46:51,040 --> 00:46:53,200 IN THE ACOUSTIC WORLD THAT'S 1134 00:46:53,200 --> 00:46:54,840 PROBABLY A DISADVANTAGE. 1135 00:46:54,840 --> 00:46:56,400 >>THAT'S A GOOD POINT. 1136 00:46:56,400 --> 00:46:59,480 JUST ONE LAST QUESTION, WHEN YOU 1137 00:46:59,480 --> 00:47:02,120 ARE DOING YOUR SERIAL SOLUTION 1138 00:47:02,120 --> 00:47:04,120 WITH THE ECHO DO YOU BACK FILL 1139 00:47:04,120 --> 00:47:06,840 WITH DSMO TO KEEP IT CONSTANT 1140 00:47:06,840 --> 00:47:10,040 WITH THE ECHO AS WELL? 1141 00:47:10,040 --> 00:47:11,960 >>IT DEPENDS ON THE SYSTEM 1142 00:47:11,960 --> 00:47:16,840 WE'RE USING AND THE VOLUME 1143 00:47:16,840 --> 00:47:19,000 REQUIRED FOR THE BACK FILL SO 1144 00:47:19,000 --> 00:47:20,200 DEPENDING ON THE DELIVERABLE 1145 00:47:20,200 --> 00:47:22,760 VOLUME IT MAY BE AN EX POLICE 1146 00:47:22,760 --> 00:47:29,480 AN EXPLICITDMSO PLATE THAT GET E 1147 00:47:29,480 --> 00:47:31,680 ECHO TO DO THOSE BACK FILLS OR 1148 00:47:31,680 --> 00:47:34,280 IF IT'S A HIGH HAVE IT MAY BE 1149 00:47:34,280 --> 00:47:38,600 DONE WITH MULTI-DROP COM BNL OR 1150 00:47:38,600 --> 00:47:43,840 OTHER SOME NANO LEADER DISPEN 1151 00:47:43,840 --> 00:47:44,120 DISPENSER. 1152 00:47:44,120 --> 00:47:48,760 >>I KNOW THEY HAVE BEEN THIS 1153 00:47:48,760 --> 00:47:50,240 CONCERN AROUND THE HIGHEST 1154 00:47:50,240 --> 00:47:53,400 CONCENTRATION OF THE DMSO TO USE 1155 00:47:53,400 --> 00:47:56,320 FOR CELL BASED ASSAYS AND DO YOU 1156 00:47:56,320 --> 00:47:58,840 HAVE ANY SUGGESTION ON THAT? 1157 00:47:58,840 --> 00:48:00,920 >>MOST PEOPLE DEFAULT TO NOT 1158 00:48:00,920 --> 00:48:04,480 GOING HIGHER THAN .5% DMSO IN 1159 00:48:04,480 --> 00:48:05,680 CELL BASED AND I KNOW SOME, 1160 00:48:05,680 --> 00:48:08,040 WE'VE SEEN DATA WHERE THEY CAN 1161 00:48:08,040 --> 00:48:11,720 TOLERATE UP TO 1.5% BUT WE 1162 00:48:11,720 --> 00:48:12,920 USUALLY RECOMMEND ALWAYS LESS 1163 00:48:12,920 --> 00:48:14,720 THAN 1% AND CERTAINLY THAT'S AN 1164 00:48:14,720 --> 00:48:17,120 EXPERIMENT THAT YOU CAN RUN 1165 00:48:17,120 --> 00:48:17,960 BECAUSE THERE'S DIFFERENCES 1166 00:48:17,960 --> 00:48:18,760 BETWEEN THE CELL LINE. 1167 00:48:18,760 --> 00:48:20,040 >>EXACTLY, YEAH. 1168 00:48:20,040 --> 00:48:21,800 >>I KNOW THE DEFAULT WITHOUT 1169 00:48:21,800 --> 00:48:23,080 DOING THAT EXPERIMENT IS TRYING 1170 00:48:23,080 --> 00:48:30,080 TO GET IT TO .5% FINAL IN THE 1171 00:48:30,080 --> 00:48:30,280 ASSAY. 1172 00:48:30,280 --> 00:48:32,120 >>THANK YOU. 1173 00:48:32,120 --> 00:48:37,840 [APPLAUSE] 1174 00:48:37,840 --> 00:48:39,360 >>THANK YOU, VERY MUCH, TRAVIS 1175 00:48:39,360 --> 00:48:41,040 AND LANE FOR A WONDERFUL 1176 00:48:41,040 --> 00:48:42,440 PRESENTATION AND FOR YOUR TIME 1177 00:48:42,440 --> 00:48:42,640 TODAY. 1178 00:48:42,640 --> 00:48:43,840 ALL RIGHT, WE ARE GOING TO MOVE 1179 00:48:43,840 --> 00:48:46,000 ONTO THE NEXT TALK. 1180 00:48:46,000 --> 00:48:49,160 OUR NEXT SPEAKER IS Dr. 1181 00:48:49,160 --> 00:48:49,680 MICHELLE ARKIN. 1182 00:48:49,680 --> 00:48:51,560 MICHELLE IS A PROFESSOR OF CHEM 1183 00:48:51,560 --> 00:48:53,520 TREE AT THE UNIVERSITY OF 1184 00:48:53,520 --> 00:48:58,000 CALIFORNIA SAN FRANCISCO AND 1185 00:48:58,000 --> 00:49:02,160 ADJUNCT PROFESSOR AT THE BUCK 1186 00:49:02,160 --> 00:49:04,000 INSTITUTE OF RESEARCH. 1187 00:49:04,000 --> 00:49:06,160 CHEMICAL OF PROTEIN INTERACTION 1188 00:49:06,160 --> 00:49:07,800 NETWORKS AND OTHER CHALLENGING 1189 00:49:07,800 --> 00:49:09,800 TARGETS IN DISEASES SUCH AS 1190 00:49:09,800 --> 00:49:11,960 ALZHEIMER'S AND CANCER. 1191 00:49:11,960 --> 00:49:15,080 MICHELLE ALSO CO DIRECTS THE 1192 00:49:15,080 --> 00:49:17,200 UCSF SMALL MOLECULE AND 1193 00:49:17,200 --> 00:49:18,720 EDITORIAL BOARD MEMBER OF THE 1194 00:49:18,720 --> 00:49:20,720 AGM AND MY PREVIOUS ADVISER SO 1195 00:49:20,720 --> 00:49:22,960 I'M REALLY EXCITED TO HAVE HER 1196 00:49:22,960 --> 00:49:24,080 WITH US HERE TODAY. 1197 00:49:24,080 --> 00:49:26,920 HER TALK IS ENTITLED BIO FISCAL 1198 00:49:26,920 --> 00:49:28,360 APPROACHES TO SMALL MOLECULE 1199 00:49:28,360 --> 00:49:30,400 DISCOVERY AND VALIDATION. 1200 00:49:30,400 --> 00:49:40,880 MICHELLE, THE FLOOR IS YOURS. 1201 00:49:44,440 --> 00:49:46,480 >>I LIKE THE DECISIVE BUTTON 1202 00:49:46,480 --> 00:49:46,680 PUSH. 1203 00:49:46,680 --> 00:49:50,160 LET'S SEE, I HAVE A FANCY -- 1204 00:49:50,160 --> 00:49:58,320 IT'S FANCY. 1205 00:49:58,320 --> 00:49:59,280 THERE WE GO. 1206 00:49:59,280 --> 00:50:00,720 I'M NOT SURE IF THIS IS GOING TO 1207 00:50:00,720 --> 00:50:04,600 WORK OR NOT BUT I'LL GIVE IT A 1208 00:50:04,600 --> 00:50:04,960 TRY. 1209 00:50:04,960 --> 00:50:06,480 PLEASURE TO TALK TO YOU TODAY 1210 00:50:06,480 --> 00:50:11,160 ABOUT BIO PHYSICAL STUDIES. 1211 00:50:11,160 --> 00:50:12,520 SO, WHAT I WANT TO MAKE SURE YOU 1212 00:50:12,520 --> 00:50:14,120 UNDERSTAND IS WHAT INFORMATION 1213 00:50:14,120 --> 00:50:19,080 DIFFERENT BIO FISCAL CAN 1214 00:50:19,080 --> 00:50:19,280 PROVIDE. 1215 00:50:19,280 --> 00:50:21,080 IF THERE ARE NEW METHODOLOGIES 1216 00:50:21,080 --> 00:50:22,120 YOU'VE HEARD ABOUT THAT I DON'T 1217 00:50:22,120 --> 00:50:30,400 TALK ABOUT, PLEASE, FEEL FREE TO 1218 00:50:30,400 --> 00:50:31,280 ASK. 1219 00:50:31,280 --> 00:50:34,360 IF WE HAVE TIME WE CAN USE 1220 00:50:34,360 --> 00:50:35,640 FRAGMENT-BASED DISCOVERY WITH 1221 00:50:35,640 --> 00:50:39,160 SAY APPLICATION FOR BIS OWE 1222 00:50:39,160 --> 00:50:40,680 PHYSICAL FOR PRIMARY SCREENING. 1223 00:50:40,680 --> 00:50:42,880 >>GOOD-BYE. 1224 00:50:42,880 --> 00:50:43,280 [LAUGHTER] 1225 00:50:43,280 --> 00:50:45,840 >>THAT'S THE SHORTEST TALK I'VE 1226 00:50:45,840 --> 00:50:46,240 GIVEN. 1227 00:50:46,240 --> 00:50:49,040 IT TAKES PEOPLE A FEW MINUTES 1228 00:50:49,040 --> 00:50:51,040 BEFORE THEY BOO ME OFF THE 1229 00:50:51,040 --> 00:50:51,240 STAGE. 1230 00:50:51,240 --> 00:50:51,640 [LAUGHTER] 1231 00:50:51,640 --> 00:50:54,480 SO THE GUIDING PRINCIPLE OF DRUG 1232 00:50:54,480 --> 00:50:55,640 DISCOVERY IT'S IMPLICIT IN 1233 00:50:55,640 --> 00:50:57,440 EVERYTHING THAT ARE DOING BUT 1234 00:50:57,440 --> 00:50:59,560 BIOPHYSICS MAKES IT EXPLICIT SO 1235 00:50:59,560 --> 00:51:01,600 THE GOAL OF DRUG DISCOVERY IS TO 1236 00:51:01,600 --> 00:51:03,640 FIND A MOLECULE THAT MODULATES 1237 00:51:03,640 --> 00:51:04,960 THE FUNCTION OF SOMETHING 1238 00:51:04,960 --> 00:51:06,640 USUALLY A PROTEIN BY BINDING TO 1239 00:51:06,640 --> 00:51:10,600 A SINGLE CORRECT SITE ON THAT 1240 00:51:10,600 --> 00:51:11,920 TARGET. 1241 00:51:11,920 --> 00:51:14,280 MOST OF THE AS SAYS DO NOT DO 1242 00:51:14,280 --> 00:51:16,800 THAT. THEY SHOW INHIBITION OF A 1243 00:51:16,800 --> 00:51:19,840 TARGET OR MODULATION OF A 1244 00:51:19,840 --> 00:51:20,800 PHENOTYPE BUT BIS OWE PHYSICAL 1245 00:51:20,800 --> 00:51:21,920 METHODS CAN TELL YOU THAT YOU 1246 00:51:21,920 --> 00:51:23,800 ARE DOING THAT BY BINDING TO A 1247 00:51:23,800 --> 00:51:28,320 SINGLE TARGET AT A SINGLE SITE. 1248 00:51:28,320 --> 00:51:30,720 SO THAT IS SUGGESTS WE WANT A 1249 00:51:30,720 --> 00:51:32,560 ONE TO ONE WITH THE BINDING SITE 1250 00:51:32,560 --> 00:51:34,480 AND THE MOL I'M AND THAT ALLOWS 1251 00:51:34,480 --> 00:51:37,160 US TO FIND THE MECHANISM OF HOW 1252 00:51:37,160 --> 00:51:39,240 WE'RE MODULATING THAT PROTEIN OR 1253 00:51:39,240 --> 00:51:43,520 THAT BIO MOLECULE IS A CO VEIL 1254 00:51:43,520 --> 00:51:47,200 APARTMAND THEN IF YOU HAVE AN EE 1255 00:51:47,200 --> 00:51:49,240 IS IT BINDING COMPETITIVELY WITH 1256 00:51:49,240 --> 00:51:51,720 YOUR SUBSTRATE OR NON 1257 00:51:51,720 --> 00:51:53,840 COMPETITIVELY OR UNCOMPETITIVELY 1258 00:51:53,840 --> 00:51:59,280 SO UNCOMPETITIVE IS A BACK WATER 1259 00:51:59,280 --> 00:52:02,520 BUT A LOT OF NET ABOLISHES 1260 00:52:02,520 --> 00:52:04,920 FUNCTION THIS WAY AND WE FIND 1261 00:52:04,920 --> 00:52:06,560 COMPOUNDS THAT WORK UM 1262 00:52:06,560 --> 00:52:07,880 COMPETITIVELY AND THEY BIND TO 1263 00:52:07,880 --> 00:52:11,200 THE ENZYME SUBSTRATE COMPLEX SO 1264 00:52:11,200 --> 00:52:13,040 IF YOU HAVE MORE SUBSTRATE YOU 1265 00:52:13,040 --> 00:52:14,640 HAVE TIGHTER BINDING OF YOUR 1266 00:52:14,640 --> 00:52:16,440 COMPOUND SO THESE ARE EASY 1267 00:52:16,440 --> 00:52:18,000 THINGS TO MEASURE AND YOU CAN 1268 00:52:18,000 --> 00:52:21,680 LEARN HOW YOUR DRUG IS WORKING. 1269 00:52:21,680 --> 00:52:23,800 SO, AS I MENTIONED, MOST 1270 00:52:23,800 --> 00:52:28,160 HIGH-THROUGHPUT JONES WHICH IS N 1271 00:52:28,160 --> 00:52:30,160 FUNCTION AND VIRTUAL SCREENING 1272 00:52:30,160 --> 00:52:32,280 FOCUSES ON VIRTUAL BINDING SO 1273 00:52:32,280 --> 00:52:35,480 THAT IS A NATURAL NEXT STEP IS 1274 00:52:35,480 --> 00:52:36,680 TO MEASURE REAL BINDING WITH 1275 00:52:36,680 --> 00:52:38,080 THOSE COMPOUNDS YOU'VE 1276 00:52:38,080 --> 00:52:42,600 IDENTIFIED VIRTUALLY. 1277 00:52:42,600 --> 00:52:44,160 SO, BIO PHYSICAL ASSAYS ALLOW 1278 00:52:44,160 --> 00:52:48,280 YOU, DEPENDING ON THE ASSAY, TO 1279 00:52:48,280 --> 00:52:49,720 MEASURE AFFINITY AND SOMETHING 1280 00:52:49,720 --> 00:52:51,360 ABOUT MECHANISM AND FOR THAT 1281 00:52:51,360 --> 00:52:53,000 REASON THEY CAN BE USED AT EACH 1282 00:52:53,000 --> 00:52:55,040 STAGE OF THE DRUG DISCOVERY 1283 00:52:55,040 --> 00:52:58,960 PROCESS AND IN TARGET 1284 00:52:58,960 --> 00:53:02,280 IDENTIFICATION, HIT EYE 1285 00:53:02,280 --> 00:53:04,280 IDENTIFICATION, WHERE YOU CAN 1286 00:53:04,280 --> 00:53:05,840 SCREEN UP TO TENS OF THOUSANDS 1287 00:53:05,840 --> 00:53:07,320 OF MOLECULES WITH 1288 00:53:07,320 --> 00:53:09,080 HIGH-THROUGHPUT VERSIONS OF 1289 00:53:09,080 --> 00:53:11,000 THOSE TECHNOLOGIES, HIT TO LEAD 1290 00:53:11,000 --> 00:53:12,800 WHERE WE HAVE A SET OF COMPOUNDS 1291 00:53:12,800 --> 00:53:15,760 AND REIMPROVE THE AFFINITY OF 1292 00:53:15,760 --> 00:53:17,000 THOSE COMPOUNDS WE WANT TO MICK 1293 00:53:17,000 --> 00:53:18,160 SURE WE'RE STAYING ON MECHANISM 1294 00:53:18,160 --> 00:53:23,000 AND THEN EVEN IN LEAD ON TOM 1295 00:53:23,000 --> 00:53:26,200 EYESIZATION WITH PEOPLE FOCUS ON 1296 00:53:26,200 --> 00:53:28,240 AFFINITY AND SELECTIVITY BUT 1297 00:53:28,240 --> 00:53:31,800 PEOPLE SAY DON'T FORGET ABOUT 1298 00:53:31,800 --> 00:53:33,680 EFFECTS REQUIRED BINDING SO IF 1299 00:53:33,680 --> 00:53:35,080 YOU CAN BIND LONGER TO YOUR 1300 00:53:35,080 --> 00:53:36,880 TARGET, YOU CAN HAVE A BETTER 1301 00:53:36,880 --> 00:53:40,120 EFFECT EVEN IF YOU ARE PHARMA CO 1302 00:53:40,120 --> 00:53:41,920 KINETICS, WHEN IT GETS CLEARED 1303 00:53:41,920 --> 00:53:43,840 IF IT'S BINDING TO YOUR TARGET 1304 00:53:43,840 --> 00:53:45,520 IT CAN BE WORKING. 1305 00:53:45,520 --> 00:53:55,840 IT'S EVEN USED IN LEAD ON TOM 1306 00:53:55,840 --> 00:53:56,600 OPTIMIZIZATION. 1307 00:53:56,600 --> 00:53:58,000 YOU HEARD FROM DOUG THAT 1308 00:53:58,000 --> 00:53:59,640 TERRIBLE THINGS CAN HAPPEN IN 1309 00:53:59,640 --> 00:54:02,360 HIGH-THROUGHPUT SCREENING. 1310 00:54:02,360 --> 00:54:04,040 SO, HIT VALIDATION, THE GOAL OF 1311 00:54:04,040 --> 00:54:06,440 THE HIT VALIDATION IS SEPARATING 1312 00:54:06,440 --> 00:54:08,880 THE WHEAT FROM THE CHAFF. 1313 00:54:08,880 --> 00:54:11,120 HE SAID THIS TO YOU, IT'S TO 1314 00:54:11,120 --> 00:54:13,640 REMOVE THE 99% OF THINGS YOU ARE 1315 00:54:13,640 --> 00:54:14,600 NOT INTERESTED IN. 1316 00:54:14,600 --> 00:54:16,120 AND THEN THE NEXT STEP OF 1317 00:54:16,120 --> 00:54:19,080 VALIDATION IS TO SEE WHAT 1% OF 1318 00:54:19,080 --> 00:54:21,760 THE 1% OF THINGS THAT YOU GOT 1319 00:54:21,760 --> 00:54:23,160 ARE INTERESTING. 1320 00:54:23,160 --> 00:54:25,640 SO, IF YOU HAVE A LOT OF ASSAY 1321 00:54:25,640 --> 00:54:27,880 INTERFERENCE OR A LOT OF 1322 00:54:27,880 --> 00:54:29,200 SATURDAY FACTS IN YOUR ASSAY 1323 00:54:29,200 --> 00:54:31,040 IT'S HARD TO PULL OUT THOSE THAT 1324 00:54:31,040 --> 00:54:33,320 ARE REAL SO WE TRIED TO DEVELOP 1325 00:54:33,320 --> 00:54:34,400 OUR ASSAY SO WE HAVE A FEW 1326 00:54:34,400 --> 00:54:35,840 COMMON ARTIFACTS AS POSSIBLEMENT 1327 00:54:35,840 --> 00:54:37,480 FOR EXAMPLE, IF YOU HAVE 1328 00:54:37,480 --> 00:54:39,400 COMPOUND AGGREGATION USING 1329 00:54:39,400 --> 00:54:42,280 DETERGENT AND CARRIER PROTEINS 1330 00:54:42,280 --> 00:54:42,840 AND IRRELEVANT PROTEINS CAN 1331 00:54:42,840 --> 00:54:44,240 REDUCE THE ARTIFACTS THAT YOU 1332 00:54:44,240 --> 00:54:46,720 SEE IN THE FIRST PLACE BUT YOU 1333 00:54:46,720 --> 00:54:48,240 STILL NEED TO UNDERSTAND THAT 1334 00:54:48,240 --> 00:54:49,760 YOU WILL PROBABLY HAVE ARTIFACTS 1335 00:54:49,760 --> 00:54:51,960 IN YOUR SCREEN AND THIS CAN BE 1336 00:54:51,960 --> 00:54:54,280 DO YOU TO ASSAY INTERFERENCE AND 1337 00:54:54,280 --> 00:54:56,120 METRIC INTERFERENCE AND THE 1338 00:54:56,120 --> 00:54:58,760 COMPOUND AGGREGATION THAT IS COM 1339 00:54:58,760 --> 00:55:00,360 OND AND CHEMICAL REACTIVITY WE 1340 00:55:00,360 --> 00:55:02,440 CAN SEE FROM THE STRUCTURE AND 1341 00:55:02,440 --> 00:55:04,480 WE'LL HAVE FLAGS ABOUT WHAT 1342 00:55:04,480 --> 00:55:06,320 COMPOUNDS COULD BE CO VAL ANT OR 1343 00:55:06,320 --> 00:55:10,560 REACTIVE BUT YOU CAN STILL BE 1344 00:55:10,560 --> 00:55:12,840 SURPRISED AND IF YOU CAN HAVE 1345 00:55:12,840 --> 00:55:15,040 REVERS ABLE BINDING AND YOU CAN 1346 00:55:15,040 --> 00:55:24,640 FIGURE OUT IS IT COVALANT BUT IF 1347 00:55:24,640 --> 00:55:26,440 YOU CAN PROVE THE POSITIVE, THAT 1348 00:55:26,440 --> 00:55:28,960 YOUR COMPOUND IS BINDING 1349 00:55:28,960 --> 00:55:30,400 REVERSIBLY ONE TO ONE BINDING AT 1350 00:55:30,400 --> 00:55:35,400 A SINGLE CITE IT ELIMINATES THE 1351 00:55:35,400 --> 00:55:35,680 ARTIFACTS. 1352 00:55:35,680 --> 00:55:37,200 SO WE HAVE A LOT OF DIFFERENT 1353 00:55:37,200 --> 00:55:39,000 ASSAYS WE CAN USE AND DEPENDING 1354 00:55:39,000 --> 00:55:41,440 ON THE SIZE AND EXPERTISE OF THE 1355 00:55:41,440 --> 00:55:42,440 LABORATORIES, YOUR LAB AND THE 1356 00:55:42,440 --> 00:55:43,880 LABS AROUND YOU, YOU MAY HAVE 1357 00:55:43,880 --> 00:55:46,240 ACCESS TO A LOT OF THESE. 1358 00:55:46,240 --> 00:55:47,840 SO, YOU START WITH THINKING, 1359 00:55:47,840 --> 00:55:50,160 WHAT DO I WANT TO DEMONSTRATORS 1360 00:55:50,160 --> 00:55:52,080 WHAT DO I NOT KNOW AND WHAT 1361 00:55:52,080 --> 00:55:52,680 ASSAY CAN I USE? 1362 00:55:52,680 --> 00:55:54,200 THESE ARE SOME OF THE PROPERTIES 1363 00:55:54,200 --> 00:55:57,600 OF INHIBITOR THAT YOU MAY BE 1364 00:55:57,600 --> 00:55:58,120 CONCERNED ABOUT. 1365 00:55:58,120 --> 00:55:59,880 THESE ARE VARIOUS METHOD ANOTHER 1366 00:55:59,880 --> 00:56:01,280 GEES AND THIS IS MEANT AS A 1367 00:56:01,280 --> 00:56:03,040 SLIDE FOR YOU, A TABLE FOR YOU 1368 00:56:03,040 --> 00:56:04,360 TO GO BACK TO BUT I'LL GO 1369 00:56:04,360 --> 00:56:07,800 THROUGH A FEW OF THE MOST COMMON 1370 00:56:07,800 --> 00:56:08,040 ONES. 1371 00:56:08,040 --> 00:56:10,320 SO I'LL START WITH LIGHTS 1372 00:56:10,320 --> 00:56:19,080 GATHERING AND WITH DIFFERENTIAL 1373 00:56:19,080 --> 00:56:19,960 SCANNING. 1374 00:56:19,960 --> 00:56:23,360 IT'S A WAY OF IDENTIFYING 1375 00:56:23,360 --> 00:56:26,240 PROTEIN AGO AGREE GATORS, 1376 00:56:26,240 --> 00:56:30,040 COMPOUNDS AGGREGATE INTO THESE 1377 00:56:30,040 --> 00:56:33,800 STRUCTURES THAT ARE SOL YOU ABLE 1378 00:56:33,800 --> 00:56:35,640 AND TRANSPARENT. 1379 00:56:35,640 --> 00:56:37,720 CAN'T SPIN THEM OUT AND YOU 1380 00:56:37,720 --> 00:56:40,000 CAN'T SEE THEM. 1381 00:56:40,000 --> 00:56:41,760 ONE THING THAT I REALLY WANT TO 1382 00:56:41,760 --> 00:56:43,560 CALL YOUR ATTENTION TO IS IN THE 1383 00:56:43,560 --> 00:56:47,080 EARLY DAYS OF THINKING ABOUT 1384 00:56:47,080 --> 00:56:49,600 AGGREGATORS, THIS COM SOUND IS 1385 00:56:49,600 --> 00:56:52,320 AN AGGREGATORS BUT OTHER DRUGS 1386 00:56:52,320 --> 00:56:55,760 ARE AGO AGREE GATORS TOO UNDER 1387 00:56:55,760 --> 00:56:56,240 CIRCUMSTANCES. 1388 00:56:56,240 --> 00:56:59,440 THE AGGREGATION DEPENDS ON THE 1389 00:56:59,440 --> 00:57:01,680 PROTEIN, IT DEPENDS ON THE 1390 00:57:01,680 --> 00:57:03,080 BUFFER CONDITIONS AND 1391 00:57:03,080 --> 00:57:04,640 CONCENTRATION DEPENDENT ALSO SO 1392 00:57:04,640 --> 00:57:07,040 IT'S SOMETHING WE CAN MEASURE. 1393 00:57:07,040 --> 00:57:09,680 MEASURE THE CONCENTRATION 1394 00:57:09,680 --> 00:57:13,120 DEPENDENCE OF THE FORMATION OF 1395 00:57:13,120 --> 00:57:15,280 THESE SOL YOU ABLE ARTICLE THE 1396 00:57:15,280 --> 00:57:17,200 COMPOUND WILL AGGREGATE AND IT'S 1397 00:57:17,200 --> 00:57:19,040 NOT A BIG DEAL THE ISSUE IS YOU 1398 00:57:19,040 --> 00:57:20,960 WANT IT NOT TO AGGREGATE WITH 1399 00:57:20,960 --> 00:57:25,960 THE SAME EC50 OR THE SAME 1400 00:57:25,960 --> 00:57:27,040 CONCENTRATION AS THE ACTIVITY 1401 00:57:27,040 --> 00:57:27,840 YOU ARE FINDING. 1402 00:57:27,840 --> 00:57:30,000 YOU WANT TO DEMONSTRATION THE 1403 00:57:30,000 --> 00:57:33,160 AGGREGATION IS NOT THE CAUSE OF 1404 00:57:33,160 --> 00:57:33,920 YOUR FUNCTIONAL ASSAY. 1405 00:57:33,920 --> 00:57:38,920 AND THIS IS WHAT THE DATA LOOKED 1406 00:57:38,920 --> 00:57:41,760 LIKE AND AGGREGATE WILL HAVE 1407 00:57:41,760 --> 00:57:43,680 THIS LONG CORRELATION TIME AND 1408 00:57:43,680 --> 00:57:46,560 THEN WE FIT THIS, YOU JUST GET A 1409 00:57:46,560 --> 00:57:49,680 NUMBER FOR CORRELATION TIME AND 1410 00:57:49,680 --> 00:57:51,560 THAT Y AXIS TIME. 1411 00:57:51,560 --> 00:57:53,120 YOU CAN LOCK AT THAT AS A 1412 00:57:53,120 --> 00:57:54,720 FUNCTION OF CONCENTRATION AND 1413 00:57:54,720 --> 00:57:57,040 PLOT THAT VERSUS YOUR IC50 AND 1414 00:57:57,040 --> 00:57:58,960 JUST MAKE SURE THEY DON'T CROSS 1415 00:57:58,960 --> 00:58:00,960 EACH OTHER. 1416 00:58:00,960 --> 00:58:10,280 N THEY'RE JUST ROTATING FREELY N 1417 00:58:10,280 --> 00:58:13,520 SOLUTION. 1418 00:58:13,520 --> 00:58:14,720 AGGREGATION SO IT WILL MEASURE 1419 00:58:14,720 --> 00:58:16,400 THE RADIUS, WE DON'T USUALLY 1420 00:58:16,400 --> 00:58:18,520 LOCK AT RADIUS WE JUST LOOK AT 1421 00:58:18,520 --> 00:58:26,280 THE PRESENCE OF AGRI GATES. 1422 00:58:26,280 --> 00:58:28,680 IT TENDS TO BE A STEEP 1423 00:58:28,680 --> 00:58:29,800 CONCENTRATION CURVE LIKE MY CELL 1424 00:58:29,800 --> 00:58:34,240 IF YOU WORK IN MY C YOU CELLS. 1425 00:58:34,240 --> 00:58:36,200 YOU SEE NOTHING AND NOTHING AND 1426 00:58:36,200 --> 00:58:38,560 A BIG STEEP AND THEN IT'S HIGH 1427 00:58:38,560 --> 00:58:40,120 AND CONSTANT USUALLY. 1428 00:58:40,120 --> 00:58:43,920 SO THESE INSTRUMENTS YOU CAN GET 1429 00:58:43,920 --> 00:58:45,520 IN 384 FORMAT NOW AND THEY CAN 1430 00:58:45,520 --> 00:58:47,120 BE DONE VERY QUICKLY AND THE 1431 00:58:47,120 --> 00:58:50,520 TRICK IS, NOT HAVING ANY MY 1432 00:58:50,520 --> 00:58:52,080 CELLS OR BIG THINGS IN YOUR 1433 00:58:52,080 --> 00:58:53,600 BUFFER IN THE FIRST PLACE SO 1434 00:58:53,600 --> 00:58:57,520 USUALLY WHEN YOU ARE DOING A DLS 1435 00:58:57,520 --> 00:58:58,720 EXPERIMENT, YOU WOULDN'T HAVE 1436 00:58:58,720 --> 00:59:05,640 CARRIER PROTEINS OR DETER GANTZ 1437 00:59:05,640 --> 00:59:06,480 AND YOU ARE NEED TO MAKE SURE 1438 00:59:06,480 --> 00:59:09,360 YOU HAVE NO DUST SO THAT IS 1439 00:59:09,360 --> 00:59:11,080 REALLY THE TRICK TO THIS IS JUST 1440 00:59:11,080 --> 00:59:12,360 DUST FREE AND THEN IT'S A 1441 00:59:12,360 --> 00:59:19,720 STRAIGHT FORWARD TECHNIQUE. 1442 00:59:19,720 --> 00:59:21,480 ONE THING A LOT OF PEOPLE WILL 1443 00:59:21,480 --> 00:59:24,360 DO BECAUSE IT YOU DO IT IN 384 1444 00:59:24,360 --> 00:59:27,080 FORMAT STRAIGHT FORWARDLY IS THE 1445 00:59:27,080 --> 00:59:31,960 PRINCIPLE BEHIND TERMAL DEN 1446 00:59:31,960 --> 00:59:38,360 ATURATION, IT'S MORE STABLE THAN 1447 00:59:38,360 --> 00:59:48,920 THE UNBOUND AND IF YOU PUT IT IO 1448 00:59:56,440 --> 01:00:05,160 A RTCPR SYSTEM AND YOU ADD A DYE 1449 01:00:05,160 --> 01:00:07,400 THAT BINDS TO THE FORM OF A PRE 1450 01:00:07,400 --> 01:00:09,400 TEEN SO WHEN A PROTEIN STARTS TO 1451 01:00:09,400 --> 01:00:15,040 MELT AND HYDRO PHOBIC SURFACES 1452 01:00:15,040 --> 01:00:19,240 BIND AND IT GIVES A INCREASE IN 1453 01:00:19,240 --> 01:00:19,960 FLUORESCENCE SIGNAL FROM 1454 01:00:19,960 --> 01:00:20,360 TEMPERATURE. 1455 01:00:20,360 --> 01:00:22,800 FROM THE MIDDLE GRAPH, YOU CAN 1456 01:00:22,800 --> 01:00:24,560 SEE THAT WE HAVE A MELTING 1457 01:00:24,560 --> 01:00:29,600 TEMPERATURE FOR THIS PROTEIN OF 1458 01:00:29,600 --> 01:00:32,800 ABOUT 60° AND AS AS YOU 1459 01:00:32,800 --> 01:00:34,480 INCREASE, YOU SEE FLUORESCENCE 1460 01:00:34,480 --> 01:00:38,240 IE YOU START TO SEE PROTEIN 1461 01:00:38,240 --> 01:00:39,200 DENATURATION AT HIGHER AND 1462 01:00:39,200 --> 01:00:41,040 HIGHER TEMPERATURES AND FROM 1463 01:00:41,040 --> 01:00:43,680 THIS YOU CAN MUR KD FOR THE 1464 01:00:43,680 --> 01:00:45,000 COMPOUND AND YOU CAN 1465 01:00:45,000 --> 01:00:46,040 DEMONSTRATION THAT THE COMPOUND 1466 01:00:46,040 --> 01:00:48,040 IS BINDING TO THE PROTEIN 1467 01:00:48,040 --> 01:00:50,240 USUALLY AGO AGREE GATERS WILL 1468 01:00:50,240 --> 01:00:51,640 NOT INCREASE THE STABILITY OF 1469 01:00:51,640 --> 01:01:01,720 YOUR PROTEIN. 1470 01:01:01,720 --> 01:01:03,600 WHEN YOU ADD YOUR COMPOUND. 1471 01:01:03,600 --> 01:01:04,840 THIS MEANS ALL KINDS OF THINGS, 1472 01:01:04,840 --> 01:01:06,520 PEOPLE TEND NOT TO WORRY ABOUT 1473 01:01:06,520 --> 01:01:06,800 IT. 1474 01:01:06,800 --> 01:01:09,680 IT COULD MEAN IT'S BINDING TO A 1475 01:01:09,680 --> 01:01:11,120 LOWER -- A LESS STABLE STATE OF 1476 01:01:11,120 --> 01:01:12,200 YOUR PROTEIN. 1477 01:01:12,200 --> 01:01:13,240 IT COULD MEAN THAT YOU ARE 1478 01:01:13,240 --> 01:01:14,880 BINDING TO THE UNFOLDED STATE OF 1479 01:01:14,880 --> 01:01:16,600 YOUR PROTEIN SO YOU HAVE TO 1480 01:01:16,600 --> 01:01:18,080 THINK ABOUT WHAT IS LIKELY FOR 1481 01:01:18,080 --> 01:01:20,120 YOUR SYSTEM. 1482 01:01:20,120 --> 01:01:24,360 ANOTHER WAY TO DO THERMAL 1483 01:01:24,360 --> 01:01:26,720 DENATURATION LOWER AND IT'S A 1484 01:01:26,720 --> 01:01:33,720 TWO CD AS YOU MELT YOUR PROTEIN 1485 01:01:33,720 --> 01:01:35,240 AND YOU LOSE YOUR SECONDARY 1486 01:01:35,240 --> 01:01:39,720 STRUCTURE AND YOU CAN SEE IT 1487 01:01:39,720 --> 01:01:40,160 CLEARLY. 1488 01:01:40,160 --> 01:01:44,400 DO I HAVE D SF 2.0. 1489 01:01:44,400 --> 01:01:48,880 SO MOSTLY DSF RIGHT NOW, IS DONE 1490 01:01:48,880 --> 01:01:50,840 WITH A DYE CALLED CYBER ORANGE 1491 01:01:50,840 --> 01:01:52,280 AND IT WORKS FOR A LOT OF 1492 01:01:52,280 --> 01:01:54,160 PROTEINS BUT NOT ALL PROTEINS SO 1493 01:01:54,160 --> 01:01:57,280 A COLLEAGUE OF MINE HAS A 1494 01:01:57,280 --> 01:01:59,880 RESEARCH PROGRAM FOCUSED ON 1495 01:01:59,880 --> 01:02:01,520 SCREENING DYE SO WE HAVE A DYE 1496 01:02:01,520 --> 01:02:02,880 LIBRARY AND WE SCREEN THEM FOR 1497 01:02:02,880 --> 01:02:04,200 SPECIFIC PROTEINS AND SO IF YOU 1498 01:02:04,200 --> 01:02:06,280 ARE COMPOUND DOESN'T, IF YOUR 1499 01:02:06,280 --> 01:02:14,880 PREIF PROTEINDOESN'T, WE TRY A E 1500 01:02:14,880 --> 01:02:15,600 AND WE ARE PLAYING WITH 1501 01:02:15,600 --> 01:02:16,920 DIFFERENT THINGS. 1502 01:02:16,920 --> 01:02:18,000 CAN YOU FIND DYES THAT BIND TO 1503 01:02:18,000 --> 01:02:20,000 DIFFERENT DOMAINS OF A MULTI 1504 01:02:20,000 --> 01:02:21,680 DOMAIN PROTEIN OR TO ONE BUT NOT 1505 01:02:21,680 --> 01:02:23,920 THE OTHER PARTNER OF A 1506 01:02:23,920 --> 01:02:25,000 PROTEIN-PROTEIN INTERACTION SO 1507 01:02:25,000 --> 01:02:26,720 THERE ARE OTHER OPTIONS BUT 1508 01:02:26,720 --> 01:02:29,880 THERE'S ONE DYE THAT'S VERY 1509 01:02:29,880 --> 01:02:30,640 TYPICALLY USED. 1510 01:02:30,640 --> 01:02:32,800 AND THE EVOLUTION OF THIS 1511 01:02:32,800 --> 01:02:33,960 TECHNIQUE INTO CELLS SOMETHING 1512 01:02:33,960 --> 01:02:39,680 YOU MAY HAVE HEARD OF OR DO 1513 01:02:39,680 --> 01:02:41,520 YOURSELF CALLED SETSA AND IT'S 1514 01:02:41,520 --> 01:02:43,560 THE SAME THAT YOU CAN MEASURE 1515 01:02:43,560 --> 01:02:45,240 BINDING TO YOUR PRE TEEN BUT 1516 01:02:45,240 --> 01:02:48,360 ALSO IN A CELL, BY MEASURING HOW 1517 01:02:48,360 --> 01:02:51,120 THERMALLY STABLE THE PROTEIN IS 1518 01:02:51,120 --> 01:02:52,000 IN THE CELL. 1519 01:02:52,000 --> 01:02:55,280 AND IT WAS ORIGINALLY DONE WITH 1520 01:02:55,280 --> 01:02:56,200 WESTERN BLOTTING SO YOU HAVE A 1521 01:02:56,200 --> 01:02:58,040 PARTICULAR PROTEIN OF INTEREST 1522 01:02:58,040 --> 01:02:59,200 AND DIFFERENT TEMPERATURES, PLUS 1523 01:02:59,200 --> 01:03:01,600 AND MINUS THE COMPOUND, AND YOU 1524 01:03:01,600 --> 01:03:04,640 SEE THAT AT ABOUT 46° YOU START 1525 01:03:04,640 --> 01:03:06,240 TO LOSE THE PROTEIN IN THE 1526 01:03:06,240 --> 01:03:08,200 ABSENCE OF COMPOUND AND THE 1527 01:03:08,200 --> 01:03:09,920 PROTEIN IS STILL STABLE WHEN YOU 1528 01:03:09,920 --> 01:03:12,080 HEAT THE WHOLE CELL UP AND EVEN 1529 01:03:12,080 --> 01:03:13,360 UP TO 56°. 1530 01:03:13,360 --> 01:03:15,040 SO THERE'S A LARGE SHIFT IN THE 1531 01:03:15,040 --> 01:03:21,080 THERMAL STABILITY OF THE PROTEIN 1532 01:03:21,080 --> 01:03:23,920 IN CELL LIE SATES WHEN YOU HEAT 1533 01:03:23,920 --> 01:03:25,680 UP THE CELL AND THEN CAN YOU DO 1534 01:03:25,680 --> 01:03:27,440 THIS AS A FUNCTION OF 1535 01:03:27,440 --> 01:03:29,680 CONCENTRATION TO GET AN IDEA FOR 1536 01:03:29,680 --> 01:03:32,640 THE BINDING AND KD FOR BINDING 1537 01:03:32,640 --> 01:03:33,840 TO THAT PROTEIN. 1538 01:03:33,840 --> 01:03:35,280 THERE ARE MANY REASONS THAT THAT 1539 01:03:35,280 --> 01:03:36,120 MIGHT NOT WORK. 1540 01:03:36,120 --> 01:03:38,160 BUT IT SEEMS TO WORK PRETTY 1541 01:03:38,160 --> 01:03:38,680 OFTEN. 1542 01:03:38,680 --> 01:03:41,000 IT'S REMARKABLY USEFUL 1543 01:03:41,000 --> 01:03:42,800 TECHNIQUES AND THE LATEST 1544 01:03:42,800 --> 01:03:43,880 INCARNATION OF THIS TECHNOLOGY 1545 01:03:43,880 --> 01:03:47,080 IS TO USE MASS SPECTORMETRY SO 1546 01:03:47,080 --> 01:03:49,360 NOW IF YOU HAVE A CELL THAT 1547 01:03:49,360 --> 01:03:51,160 YOU'VE LABELED SAY WITH LIGHT 1548 01:03:51,160 --> 01:03:53,040 MEDIA, SO SAY YOU DO A 1549 01:03:53,040 --> 01:03:55,400 EXPERIMENT SO YOU HAVE LIGHT 1550 01:03:55,400 --> 01:03:56,720 MEDIA, CELLS WITHOUT COMPOUND 1551 01:03:56,720 --> 01:03:59,040 AND THEN HEAVY MEDIA CELLS WITH 1552 01:03:59,040 --> 01:04:00,600 COMPOUND, YOU MIX THOSE TOGETHER 1553 01:04:00,600 --> 01:04:02,240 AND YOU CAN SEE THE PROTEINS 1554 01:04:02,240 --> 01:04:04,040 HAVE SLIGHTLY DIFFERENT MASSES 1555 01:04:04,040 --> 01:04:05,320 BECAUSE THEY'RE GREN IN 1556 01:04:05,320 --> 01:04:06,240 DIFFERENT MEDIA AND YOU CAN SEE 1557 01:04:06,240 --> 01:04:07,960 AS YOU HEAT UP THE MEDIA WHICH 1558 01:04:07,960 --> 01:04:10,520 PRE TEENS YOU NO LONGER DETECT 1559 01:04:10,520 --> 01:04:12,120 BECAUSE THEY'VE DENATURED AND 1560 01:04:12,120 --> 01:04:13,640 WHICH PROTEINS YOU ARE STILL 1561 01:04:13,640 --> 01:04:14,360 DETECTING BECAUSE THEY'RE STILL 1562 01:04:14,360 --> 01:04:16,360 IN SOLUTION AND IF THAT SHIFTS 1563 01:04:16,360 --> 01:04:19,320 OVER, THE CONCENTRATION OVER THE 1564 01:04:19,320 --> 01:04:20,520 TEMPERATURE RANGE THAT YOU ARE 1565 01:04:20,520 --> 01:04:22,520 LOOKING, THEN YOU CAN SAY THAT 1566 01:04:22,520 --> 01:04:24,600 YOU'VE STABILIZED A PROTEIN OR 1567 01:04:24,600 --> 01:04:28,800 SET OF PROTEINS WITH THE 1568 01:04:28,800 --> 01:04:30,240 COMPOUND. 1569 01:04:30,240 --> 01:04:31,320 SO, ANOTHER TECHNIQUE THAT IS 1570 01:04:31,320 --> 01:04:32,560 FAIRLY HIGH-THROUGHPUT THAT 1571 01:04:32,560 --> 01:04:35,360 PEOPLE WILL USE IN HIT 1572 01:04:35,360 --> 01:04:37,480 VALIDATION, IS CALLED MICRO 1573 01:04:37,480 --> 01:04:39,120 SCALE THERMO OR MST. 1574 01:04:39,120 --> 01:04:40,840 THE PRINCIPLE HERE IS THAT THIS 1575 01:04:40,840 --> 01:04:42,400 IS KIND OF A COOKIE TECHNIQUE. 1576 01:04:42,400 --> 01:04:45,240 SO THE IDEA IS THAT PROTEINS 1577 01:04:45,240 --> 01:04:48,840 WILL MOVE IN A THERMAL GRADIENT 1578 01:04:48,840 --> 01:04:50,960 SO YOU KNOW, THINGS WILL MOVE 1579 01:04:50,960 --> 01:04:52,360 AND THEY MOVE HOW QUICK WILL HE 1580 01:04:52,360 --> 01:04:53,240 THEY MOVE IN SOLUTION IS BASED 1581 01:04:53,240 --> 01:04:56,960 ON THEIR SIZE, SHAPE AND CHARGE. 1582 01:04:56,960 --> 01:05:00,360 SO THE WAY THIS TECHNOLOGY 1583 01:05:00,360 --> 01:05:01,760 WORKS, YOU HAVE AN ICE OWE 1584 01:05:01,760 --> 01:05:03,120 TROPIC SOLUTION SO YOU JUST HAVE 1585 01:05:03,120 --> 01:05:04,680 YOUR PROTEIN AND SOLUTION AND 1586 01:05:04,680 --> 01:05:07,320 THEN AN IR AND LIGHT COMES ON 1587 01:05:07,320 --> 01:05:09,960 AND IR LASER COMES ON AND THAT 1588 01:05:09,960 --> 01:05:12,560 SCATTERS THE PROTEINS AWAY FROM 1589 01:05:12,560 --> 01:05:14,360 THE LIGHT BUT AWAY FROM THE 1590 01:05:14,360 --> 01:05:14,840 WARMTH. 1591 01:05:14,840 --> 01:05:17,200 AND THEN YOU TURN THE LASER OFF 1592 01:05:17,200 --> 01:05:19,320 AND PROTEINS MOVE BACK SO IF THE 1593 01:05:19,320 --> 01:05:22,840 PROTEINS ARE LABELED WITH A 1594 01:05:22,840 --> 01:05:24,920 FLORA FLOOR OR LABELED WITH A 1595 01:05:24,920 --> 01:05:26,400 GREEN FLORA FLOOR, THEN YOU CAN 1596 01:05:26,400 --> 01:05:28,560 DETECT THEM MOVING BACK INTO THE 1597 01:05:28,560 --> 01:05:32,720 GRADIENT AND SO HERE THEY ARE, 1598 01:05:32,720 --> 01:05:33,280 AND THEY'RE GOING AWAY AND 1599 01:05:33,280 --> 01:05:34,640 THEY'RE STARTING TO COME BACK 1600 01:05:34,640 --> 01:05:36,280 YOU CAN SEE HOW MUCH THEY MOVE 1601 01:05:36,280 --> 01:05:41,680 AND HOW LOW THEY COME BACK AS A 1602 01:05:41,680 --> 01:05:43,840 CHANGES TO THEIR SIZE, HYDRO 1603 01:05:43,840 --> 01:05:45,680 DYNAMIC RADIUS AND CHARGE SO IF 1604 01:05:45,680 --> 01:05:47,440 YOU ADD A SMALL MOLECULE, IT 1605 01:05:47,440 --> 01:05:50,000 WORKS WELL FOR PROTEIN-PROTEIN 1606 01:05:50,000 --> 01:05:50,920 INTERACTIONS AND YOU CHANGE 1607 01:05:50,920 --> 01:05:52,480 THOSE THINGS, AND YOU CAN DETECT 1608 01:05:52,480 --> 01:05:54,560 THAT AS A FUNCTION OF 1609 01:05:54,560 --> 01:05:55,920 CONCENTRATION OF A COMPOUND. 1610 01:05:55,920 --> 01:05:57,600 SO THIS IS THE LITTLE PIECE THAT 1611 01:05:57,600 --> 01:06:00,320 YOU FIT INTO A DOSE RESPONSE. 1612 01:06:00,320 --> 01:06:07,880 TO GET A BINDING CURVE. 1613 01:06:07,880 --> 01:06:09,800 SO THOSE ARE TECHNIQUES USED 1614 01:06:09,800 --> 01:06:11,000 COMMONLY IN HIGH-THROUGHPUT AND 1615 01:06:11,000 --> 01:06:13,400 THEY CAN BE DONE FOR HUNDREDS OF 1616 01:06:13,400 --> 01:06:14,520 COMPOUNDS EASILY. 1617 01:06:14,520 --> 01:06:16,040 ANOTHER TECHNIQUE THAT IS USED 1618 01:06:16,040 --> 01:06:18,560 AT SEVERAL STAGES IN DRUG 1619 01:06:18,560 --> 01:06:20,000 DISCOVERY, SMALL MOLECULE 1620 01:06:20,000 --> 01:06:25,800 DISCOVER ROW IS SURFACE PLASMA 1621 01:06:25,800 --> 01:06:26,120 RIS INANCE. 1622 01:06:26,120 --> 01:06:28,160 IT'S A FLEXIBILITY BIO PHYSICAL 1623 01:06:28,160 --> 01:06:28,520 FORMAT. 1624 01:06:28,520 --> 01:06:30,520 IT'S CALLED A LABEL FREE FORMAT 1625 01:06:30,520 --> 01:06:31,840 WHICH IS JUST COMPLETELY NOT 1626 01:06:31,840 --> 01:06:33,120 TRUE BECAUSE THE ENTIRE 1627 01:06:33,120 --> 01:06:35,840 INSTRUMENT IS THE LABEL. 1628 01:06:35,840 --> 01:06:39,120 THE THE PROTEIN IS CLEARLY WHY 1629 01:06:39,120 --> 01:06:39,520 WRACK FOR THEM. 1630 01:06:39,520 --> 01:06:42,400 I DO USE A LOT OF SPR. 1631 01:06:42,400 --> 01:06:45,240 THE PROTEIN IS IMMOBILIZED ON TO 1632 01:06:45,240 --> 01:06:48,040 A SURFACE AND THAT'S THE PRO AND 1633 01:06:48,040 --> 01:06:49,680 CON OF THE WHOLE TECHNIQUE. 1634 01:06:49,680 --> 01:06:51,040 YOU HAVE TO BE VERY CAREFUL 1635 01:06:51,040 --> 01:06:52,960 ABOUT HOW YOUR IMMOBILIZING YOUR 1636 01:06:52,960 --> 01:06:55,200 PROTEIN AND HOW DENSELY IT'S 1637 01:06:55,200 --> 01:06:57,080 IMMOBILIZED AND SO FOURTH BUT 1638 01:06:57,080 --> 01:06:58,920 YOU IMMOBILIZE YOUR PROTEIN ON 1639 01:06:58,920 --> 01:07:02,360 THIS SURFACE AND YOU FLOW -- 1640 01:07:02,360 --> 01:07:05,600 THEY'RE CALLED LIGENS AND THEN 1641 01:07:05,600 --> 01:07:09,000 YOU FLOW AND ANNA LIGHT OR A 1642 01:07:09,000 --> 01:07:10,320 BINDING ELEMENT IN GREN OVER 1643 01:07:10,320 --> 01:07:13,000 THAT IMMOBILIZED PROTEIN. 1644 01:07:13,000 --> 01:07:18,600 WHAT SPR MEASURES IS REFRACK 1645 01:07:18,600 --> 01:07:22,360 TIVE INDEX IN THE DUFFER CLOSE 1646 01:07:22,360 --> 01:07:23,840 TO THIS SURFACE. 1647 01:07:23,840 --> 01:07:26,240 IT CHANGES THE REFLECTION OF A 1648 01:07:26,240 --> 01:07:33,720 LIGHT, LATER ON THE OPPOSITE 1649 01:07:33,720 --> 01:07:39,680 FACE. 1650 01:07:39,680 --> 01:07:42,320 IT DEPENDS ON REFRACTIVE INDEX 1651 01:07:42,320 --> 01:07:46,400 AND THE REAL MAGIC OF SPR FOR 1652 01:07:46,400 --> 01:07:47,840 SMALL MOLECULE DISCOVERY, THE 1653 01:07:47,840 --> 01:07:49,520 CHANGE IS PROPORTIONAL TO THE 1654 01:07:49,520 --> 01:07:50,920 MASS OF MATERIALS ON THE 1655 01:07:50,920 --> 01:07:51,240 SURFACE. 1656 01:07:51,240 --> 01:07:52,800 MOR OR LESS PROPORTIONAL TO MASS 1657 01:07:52,800 --> 01:07:55,000 WHICH MEANS IF YOU KNOW HOW MUCH 1658 01:07:55,000 --> 01:07:58,840 MASS YOU'VE IMMOBILIZED AND THE 1659 01:07:58,840 --> 01:08:00,400 MOLECULAR WEIGHT OF YOUR PROTEIN 1660 01:08:00,400 --> 01:08:02,680 I KNOW HOW MUCH MOLECULES YOU 1661 01:08:02,680 --> 01:08:05,040 CAN BIND IF IT'S ONE TO ONE. 1662 01:08:05,040 --> 01:08:07,120 SO THE Y AXIS TELLS YOU THE 1663 01:08:07,120 --> 01:08:08,440 NUMBER OF MOLECULES THAT ARE 1664 01:08:08,440 --> 01:08:14,800 BINDING TO THE SURFACE AND FROM 1665 01:08:14,800 --> 01:08:17,120 THAT WEIGHT YOU CAN TELL WHAT 1666 01:08:17,120 --> 01:08:20,360 THE STOICHIOMETRY. 1667 01:08:20,360 --> 01:08:21,760 THIS IS A REAL-TIME MEASUREMENT 1668 01:08:21,760 --> 01:08:24,800 SO SECONDS, NO MATERIALS AND 1669 01:08:24,800 --> 01:08:26,760 THEN YOU INJECT YOUR SAMPLE IF 1670 01:08:26,760 --> 01:08:31,040 IT DOESN'T BIND YOU HAVE NO 1671 01:08:31,040 --> 01:08:31,360 DEFLECTION. 1672 01:08:31,360 --> 01:08:33,440 IF IT BINDS, YOU SEE IT THE 1673 01:08:33,440 --> 01:08:35,120 MOLECULE BINDING TO THE 1674 01:08:35,120 --> 01:08:37,480 IMMOBILIZED PRE TEEN WITH A 1675 01:08:37,480 --> 01:08:39,160 CHARACTERISTIC CONCENTRATION 1676 01:08:39,160 --> 01:08:43,080 DEPEND ANT AND THEN YOU REACH 1677 01:08:43,080 --> 01:08:44,720 EQUILIBRIUM WHERE AS MUCH 1678 01:08:44,720 --> 01:08:49,800 COMPOUND WILL BOUN BIND AND A 1679 01:08:49,800 --> 01:08:51,280 DISASSOCIATION PHASE WHERE WHICH 1680 01:08:51,280 --> 01:08:52,800 INJECT BUFFER AND WATCH THE 1681 01:08:52,800 --> 01:08:53,680 MOLECULE COME OFF. 1682 01:08:53,680 --> 01:08:56,080 SO THIS IS NOT USUALLY WHAT SPR 1683 01:08:56,080 --> 01:08:58,080 DATA LOOKS LIKE IF YOU LOOK ON 1684 01:08:58,080 --> 01:09:00,760 THE WEB, IT'S WILL BE A SLOWER 1685 01:09:00,760 --> 01:09:03,040 ON RATE AND OFF RATE BUT FOR 1686 01:09:03,040 --> 01:09:04,240 SMALL MOLECULE DISCOVERY WHEN 1687 01:09:04,240 --> 01:09:08,640 YOU ARE OFTEN FINDING ONE 10 1688 01:09:08,640 --> 01:09:10,440 MICRO MOLAR LIGGENS IT SHOULD BE 1689 01:09:10,440 --> 01:09:12,440 FAST. 1690 01:09:12,440 --> 01:09:13,720 MOLECULE SHOULD COME ON AND OFF 1691 01:09:13,720 --> 01:09:14,120 FAST. 1692 01:09:14,120 --> 01:09:20,840 YOU CAN MEASURE THE BINDING 1693 01:09:20,840 --> 01:09:23,200 STOICHIOMETRY WITH MEASURING THE 1694 01:09:23,200 --> 01:09:24,960 MAXIMUM DISPLACEMENT WHERE 1695 01:09:24,960 --> 01:09:26,760 ADDING MORE MOLECULE HAS NO MORE 1696 01:09:26,760 --> 01:09:30,400 EFFECT ON THE SIGNAL. 1697 01:09:30,400 --> 01:09:32,800 YOU CAN MEASURE BINDING AFFINITY 1698 01:09:32,800 --> 01:09:33,720 IN TWO DIFFERENT WAYS. 1699 01:09:33,720 --> 01:09:36,560 THE YOU CAN MEASURE THE OFF RATE 1700 01:09:36,560 --> 01:09:37,880 DIVIDED BY THE ON RATE AND FIT 1701 01:09:37,880 --> 01:09:39,640 THAT GLOBAL LOW AT MULTIPLE 1702 01:09:39,640 --> 01:09:40,560 CONCENTRATIONS THAT WILL GIVE 1703 01:09:40,560 --> 01:09:42,680 YOU THE KD AND YOU CAN ALSO DO 1704 01:09:42,680 --> 01:09:47,080 MULTIPLE CONCENTRATIONS AND THE 1705 01:09:47,080 --> 01:09:50,120 MEASURE THE MAXIMUM SIGNAL AT 1706 01:09:50,120 --> 01:09:51,680 EQUILIBRIUM AS A FUNCTION OF 1707 01:09:51,680 --> 01:09:54,320 CONCENTRATION AND THAT GIVES YOU 1708 01:09:54,320 --> 01:10:00,080 A BINDING ISO THEY WERE AND 1709 01:10:00,080 --> 01:10:02,200 THEY'RE MUCH IN SINK. 1710 01:10:02,200 --> 01:10:04,720 SOME OF THE ADVANTAGES TO THIS 1711 01:10:04,720 --> 01:10:07,160 TECHNIQUE IS THAT YOU GET 1712 01:10:07,160 --> 01:10:08,880 REAL-TIME DETECTION OF BINDING 1713 01:10:08,880 --> 01:10:12,680 AND CONNETICS AND YOU CAN MRSTIK 1714 01:10:12,680 --> 01:10:16,960 ONLY TREE AND IT'S ALL ABOUT THE 1715 01:10:16,960 --> 01:10:27,440 QUALITY OF THE CHIP SURFACE. 1716 01:10:28,160 --> 01:10:33,560 I CALL THE CONVENTION SO YOU SEE 1717 01:10:33,560 --> 01:10:40,280 FINDING AND YOU ALSO SEE AGO 1718 01:10:40,280 --> 01:10:50,960 CREE GAYS, ALREGAYS -- YOU GET K 1719 01:10:50,960 --> 01:10:58,720 METRIC BINDING SO IS THE SAME 1720 01:10:58,720 --> 01:11:08,880 COMPOUND. 1721 01:11:19,200 --> 01:11:21,040 IT'S RAPID ON AND OFF WHERE IT 1722 01:11:21,040 --> 01:11:25,600 BINDS NOT SO SUPER 1723 01:11:25,600 --> 01:11:26,440 STOICHIOMETRICALLY WITH 1724 01:11:26,440 --> 01:11:32,480 CONNETICS TO JUMP TOO. 1725 01:11:32,480 --> 01:11:34,120 REMEMBER THE CAC SO IF YOU ARE 1726 01:11:34,120 --> 01:11:36,240 BELOW THE CAC IT LOOKS NORMAL 1727 01:11:36,240 --> 01:11:38,000 BUT ABOVE THE CRITICAL 1728 01:11:38,000 --> 01:11:40,000 AGGREGATION CONCENTRATION YOU 1729 01:11:40,000 --> 01:11:49,240 WILL GET HIGH ST STICK ONLY TRED 1730 01:11:49,240 --> 01:11:50,800 THIS TELLS YOU WHY YOU SHOULD 1731 01:11:50,800 --> 01:11:55,320 ALWAYS USE DETERGENT IN YOUR 1732 01:11:55,320 --> 01:12:05,680 BIOCHEMICAL AS SAYS. 1733 01:12:08,840 --> 01:12:10,560 -- THIS IS ONE WAY TO AVOID 1734 01:12:10,560 --> 01:12:12,160 SEEING ARTIFACTS AND AVOID 1735 01:12:12,160 --> 01:12:13,560 THINKING THAT REAL BINDERS ARE 1736 01:12:13,560 --> 01:12:18,920 ARTIFACTS BECAUSE YOU ARE SEEING 1737 01:12:18,920 --> 01:12:19,520 AGO AGREE GAYS THAT ISN'T 1738 01:12:19,520 --> 01:12:30,640 ENDAGO AGREE. 1739 01:12:30,880 --> 01:12:32,440 WE HAVE REACH EQUAL RA LIBERAL 1740 01:12:32,440 --> 01:12:34,280 AND WE HAVE A FAST OFF-RIGHT SO 1741 01:12:34,280 --> 01:12:36,160 WE CAN'T MENTION THE BINDING 1742 01:12:36,160 --> 01:12:38,440 CONNETICS AND WE CAN MEASURE THE 1743 01:12:38,440 --> 01:12:40,080 BINDING AFFINITY BASED ON THE 1744 01:12:40,080 --> 01:12:42,560 DOSE DEPENDENT CURVE AND WE 1745 01:12:42,560 --> 01:12:43,560 REACH SATURATION SO EACH COLOR 1746 01:12:43,560 --> 01:12:45,400 IS A DIFFERENT DOSE OF COMPOUND 1747 01:12:45,400 --> 01:12:49,240 AND IT'S DONE IN DUPLICATES AND 1748 01:12:49,240 --> 01:12:51,760 YOU REACH THE TOP, THE LIGHT 1749 01:12:51,760 --> 01:12:53,680 BLUE AND DARK BLUE ARE DIFFERENT 1750 01:12:53,680 --> 01:12:59,120 BECAUSE THEY REACH SATURATION. 1751 01:12:59,120 --> 01:13:00,440 WE HAVE COMPOUNDS THAT COME OUT 1752 01:13:00,440 --> 01:13:01,440 OF THIS SCREEN. 1753 01:13:01,440 --> 01:13:03,320 WE REPURCHASED THE HITS AND THE 1754 01:13:03,320 --> 01:13:05,960 IC50s WERE SIMILAR. 1755 01:13:05,960 --> 01:13:07,600 THE MECHANISMS OF ACTION WERE 1756 01:13:07,600 --> 01:13:09,720 SIMILAR BUT WHEN WE LOOK AT THE 1757 01:13:09,720 --> 01:13:11,320 KDs, WE SEE A BIG DIFFERENCE 1758 01:13:11,320 --> 01:13:14,440 BETWEEN THE BINDING AFFINITY 1759 01:13:14,440 --> 01:13:17,400 HERE AND THE BINDING IC50. 1760 01:13:17,400 --> 01:13:19,560 SO WHAT DO WE BELIEVE? 1761 01:13:19,560 --> 01:13:26,200 SO, THE SPC SAYS 88 MICRO MOLAR 1762 01:13:26,200 --> 01:13:27,640 ASSAY TO MICRO MOLAR WHERE THIS 1763 01:13:27,640 --> 01:13:30,080 IS MICRO MOLAR AND FOR MICRO 1764 01:13:30,080 --> 01:13:31,640 MOLAR MUCH CLOSER TO EACH OTHER. 1765 01:13:31,640 --> 01:13:36,040 WE ALSO USE DEN OMAR 1766 01:13:36,040 --> 01:13:37,320 TECHNOLOGIES BUT THIS COMPOUND 1767 01:13:37,320 --> 01:13:40,160 DIDN'T SHOW ANY SHIFT IN THE NMR 1768 01:13:40,160 --> 01:13:42,000 SO NO EVIDENCE FOR PRE TEEN TO 1769 01:13:42,000 --> 01:13:44,720 THE NMR WHERE THIS COMPOUND 1770 01:13:44,720 --> 01:13:46,840 SHOWS EVIDENCE OF BINDING BY NMR 1771 01:13:46,840 --> 01:13:52,360 SO WE HAVE TIGER WO TWO BIO PHYL 1772 01:13:52,360 --> 01:13:54,160 ASSAYS, THAT SUGGEST BINDING 1773 01:13:54,160 --> 01:13:56,880 SIMILAR TO THE CI50 BUT IT 1774 01:13:56,880 --> 01:13:58,200 DOESN'T HAVE CELL BASED ACTIVITY 1775 01:13:58,200 --> 01:13:59,760 AND WE DON'T HAVE ANY EVIDENCE 1776 01:13:59,760 --> 01:14:01,520 IT'S ACTUALLY THROUGH BINDING TO 1777 01:14:01,520 --> 01:14:03,120 THE TARGET. 1778 01:14:03,120 --> 01:14:04,920 SO YOU NEED TO PUT ALL OF THIS 1779 01:14:04,920 --> 01:14:06,280 DATA TOGETHER WHEN YOU MAKE 1780 01:14:06,280 --> 01:14:08,920 DECISIONS ABOUT WHAT TO FOLLOW. 1781 01:14:08,920 --> 01:14:10,680 SO HERE IS WHAT THE DOSE 1782 01:14:10,680 --> 01:14:14,720 RESPONSE LOOKS LIK LOCKS LIKE, L 1783 01:14:14,720 --> 01:14:17,360 THIS DATA OUT AND WE RESPONSE RU 1784 01:14:17,360 --> 01:14:19,520 AS COMPOUND CONCENTRATION AND 1785 01:14:19,520 --> 01:14:22,120 THIS GIVES US A BINDING ICE OWE 1786 01:14:22,120 --> 01:14:32,800 THEY WTHERM AND IT LOOKS LIKE A0 1787 01:14:32,800 --> 01:14:34,440 CURVE. 1788 01:14:34,440 --> 01:14:37,360 AND FROM THIS WE KNOW THAT THE 1789 01:14:37,360 --> 01:14:39,400 BMAX IS ONE EQUIVALENT SO HERE 1790 01:14:39,400 --> 01:14:43,880 IS A RULE OF THUMB, PROTEINS, 1791 01:14:43,880 --> 01:14:45,800 ABOUT 30 TO 50 TIP LIKE AND 1792 01:14:45,800 --> 01:14:49,520 SMALL MOLECULES ABOUT 300 TO 500 1793 01:14:49,520 --> 01:14:52,080 TYPICALLY SOUR LOOKING FOR A 1794 01:14:52,080 --> 01:14:54,320 1/100 RATIO AND YOU IMMOBILIZE A 1795 01:14:54,320 --> 01:14:56,600 THOUSAND RUs YOU SHOULD HAVE A 1796 01:14:56,600 --> 01:15:00,800 MAX RU OF ABOUT 10 OF YOUR SMALL 1797 01:15:00,800 --> 01:15:02,360 MOLECULE BINDING BECAUSE IT'S 1798 01:15:02,360 --> 01:15:04,840 100 THE MASS. 1799 01:15:04,840 --> 01:15:06,160 SO IN TOTAL WHAT YOU ARE LOOKING 1800 01:15:06,160 --> 01:15:09,440 FOR IN A HIT VALIDATION PACKAGE, 1801 01:15:09,440 --> 01:15:11,160 IS DEMONSTRATING REPRODUCIBILITY 1802 01:15:11,160 --> 01:15:13,040 SO YOU REPURCHASE AND YOU STILL 1803 01:15:13,040 --> 01:15:15,920 GET GOOD ACTIVITY AND THE 1804 01:15:15,920 --> 01:15:17,880 INHIBITION MECHANISM. 1805 01:15:17,880 --> 01:15:19,520 SO I DON'T THINK, I DON'T TALK 1806 01:15:19,520 --> 01:15:21,640 ABOUT MIKAELA KINETICS BUT I 1807 01:15:21,640 --> 01:15:32,320 WILL BE HAPPY TO DURING A BREAK. 1808 01:15:32,320 --> 01:15:34,360 AND BINDING DATA SO SPR GIVES 1809 01:15:34,360 --> 01:15:35,480 YOU THIS IF IT WORKS. 1810 01:15:35,480 --> 01:15:37,000 I DO TALK ABOUT IT. 1811 01:15:37,000 --> 01:15:38,200 YOU IT TALK ABOUT SOMETHING ELSE 1812 01:15:38,200 --> 01:15:42,760 BECAUSE I DO TALK ABOUT ENZYME 1813 01:15:42,760 --> 01:15:43,360 CONNETICS. 1814 01:15:43,360 --> 01:15:45,240 SO ENZYME CONNETICS, OFTEN YOU 1815 01:15:45,240 --> 01:15:47,240 WILL GET A MIXED MECHANISM WHICH 1816 01:15:47,240 --> 01:15:49,480 IS KIND OF A DRAG BECAUSE IT'S A 1817 01:15:49,480 --> 01:15:51,040 FAIRMENT OF WORK TO DO THESE SO 1818 01:15:51,040 --> 01:15:52,480 THERE ARE THREE WAYS TO 1819 01:15:52,480 --> 01:15:55,880 REPRESENT DATA SO WHAT MIKAELA 1820 01:15:55,880 --> 01:15:59,400 SMITTEN DATA MEASURES IS THE 1821 01:15:59,400 --> 01:16:01,440 EFFECT OF INHIBITION AS A 1822 01:16:01,440 --> 01:16:03,560 FUNCTION OF TIME USUALLY SO WE 1823 01:16:03,560 --> 01:16:06,360 MEASURE THE INITIAL VOLUME OS 1824 01:16:06,360 --> 01:16:07,760 TEE OF THE ENZYME REACTION AS A 1825 01:16:07,760 --> 01:16:09,960 FUNCTION OF YOUR INHIBITOR AND 1826 01:16:09,960 --> 01:16:11,520 YOUR STUB STRAIGHT. 1827 01:16:11,520 --> 01:16:13,040 SO HERE WE HAVE A LOT OF WAYS TO 1828 01:16:13,040 --> 01:16:14,360 FIT THIS DATA. 1829 01:16:14,360 --> 01:16:16,000 CONCENTRATION OF SUBSTRATE AND 1830 01:16:16,000 --> 01:16:17,520 VERSUS CONCENTRATION OF 1831 01:16:17,520 --> 01:16:18,920 COMPOUNDS SO YOU SEE THAT YOU 1832 01:16:18,920 --> 01:16:20,400 INHIBIT AS YOU ADD THE COMPOUNDS 1833 01:16:20,400 --> 01:16:25,080 YOU INHIBIT THE ASSAY. 1834 01:16:25,080 --> 01:16:28,360 THIS IS THE MODERN WAY TO FIT 1835 01:16:28,360 --> 01:16:30,520 THE DATA BECAUSE YOU ARE NOT DEG 1836 01:16:30,520 --> 01:16:31,680 ANY MANIPULATION OF THE DATA YOU 1837 01:16:31,680 --> 01:16:33,040 ARE FITTING THE DATA AS YOU 1838 01:16:33,040 --> 01:16:37,760 MEASURE IT AND YOU FIT THESE NON 1839 01:16:37,760 --> 01:16:39,760 LINEAR CURVES USING PRISM OR 1840 01:16:39,760 --> 01:16:40,160 SOMETHING. 1841 01:16:40,160 --> 01:16:43,080 THE OLD WAY IS A LIVER WEAVER 1842 01:16:43,080 --> 01:16:43,400 BURKE PLOT. 1843 01:16:43,400 --> 01:16:45,200 PEOPLE DON'T LIKE DO TO THIS 1844 01:16:45,200 --> 01:16:46,840 ANYMORE BECAUSE IT EMPHASIZES 1845 01:16:46,840 --> 01:16:48,200 CERTAIN ASPECTS OF THE CURVE 1846 01:16:48,200 --> 01:16:49,640 MORE THAN OTHERS BUT IT IS 1847 01:16:49,640 --> 01:16:52,000 REALLY NICE AS A QUICK VISUAL 1848 01:16:52,000 --> 01:16:58,240 SAYING OK, IF IT'S PARALLEL LIKE 1849 01:16:58,240 --> 01:16:59,960 THIS THIS IS AN UNCOMPETITIVE 1850 01:16:59,960 --> 01:17:01,720 MECHANISM IF IT HITS THE X AXIS 1851 01:17:01,720 --> 01:17:03,520 IT'S ONE AND Y AXIS IT'S ANOTHER 1852 01:17:03,520 --> 01:17:06,000 SO YOU CAN GET A QUICK IDEA OF 1853 01:17:06,000 --> 01:17:09,400 MECHANISM FROM LOOKING AT A LINE 1854 01:17:09,400 --> 01:17:10,440 WEAVER BURKE PLOT BUT WHAT IS 1855 01:17:10,440 --> 01:17:12,120 PRACTICAL IS TO JUST MEASURE 1856 01:17:12,120 --> 01:17:15,120 YOUR IC50 CURVE FOR YOUR SMALL 1857 01:17:15,120 --> 01:17:16,480 MOLECULE OTHERS A FUNCTION OF 1858 01:17:16,480 --> 01:17:17,920 SUBSTRATE CONCENTRATION USING 1859 01:17:17,920 --> 01:17:20,640 THE IC50 OF THE MOLECULE SO 1860 01:17:20,640 --> 01:17:22,680 HERE, I HAVE AN UNCOMPETITIVE, 1861 01:17:22,680 --> 01:17:24,800 IT'S THE SAME COMPOUND, AN 1862 01:17:24,800 --> 01:17:25,680 UNCOMPETITIVE MECHANISM OF 1863 01:17:25,680 --> 01:17:26,840 ACTION AND I KNOW THAT BECAUSE 1864 01:17:26,840 --> 01:17:30,680 IF I HAVE A LOW CONCENTRATION 6F 1865 01:17:30,680 --> 01:17:32,000 BE STRAIGHT I HAVE A WEAK IC50 1866 01:17:32,000 --> 01:17:33,440 AND 1867 01:17:33,440 --> 01:17:35,080 A HIGH 1868 01:17:35,080 --> 01:17:37,080 CONCENTRATION I HAVE A POTENT 1869 01:17:37,080 --> 01:17:39,120 IC50 SO IT GETS MORE POTENT AS 1870 01:17:39,120 --> 01:17:42,560 YOU ADD SUBSTRATE AND THAT IS 1871 01:17:42,560 --> 01:17:43,640 UNCOMPETITIVE, IT BINDS TO THE 1872 01:17:43,640 --> 01:17:45,640 ENZYME SUBSTRATE COMPLEX. 1873 01:17:45,640 --> 01:17:46,600 TYPICALLY, YOU WILL SEE 1874 01:17:46,600 --> 01:17:47,760 COMPETITIVE INHIBITION WHICH 1875 01:17:47,760 --> 01:17:49,080 WOULD MEAN THE MORE SUBSTRATE 1876 01:17:49,080 --> 01:17:50,760 YOU ADD THE WEAKER YOUR COMPOUND 1877 01:17:50,760 --> 01:17:55,520 IS SO YOU GET A LEFT -- A 1878 01:17:55,520 --> 01:17:59,640 RIGHT-WORD SHIFT IN THAT GRAPH. 1879 01:17:59,640 --> 01:18:00,440 IT GETS WEAKER. 1880 01:18:00,440 --> 01:18:01,840 THAT'S A PRACTICAL WAY TO 1881 01:18:01,840 --> 01:18:03,200 EVALUATE THE PHARMACOLOGY OF 1882 01:18:03,200 --> 01:18:04,240 YOUR COMPOUND EVEN IF YOU HAVE 1883 01:18:04,240 --> 01:18:11,360 SOME MIXED MECHANISMS BECAUSE 1884 01:18:11,360 --> 01:18:15,560 IT'S HARD TO MEASURE THESE 1885 01:18:15,560 --> 01:18:16,000 ACCURATELY. 1886 01:18:16,000 --> 01:18:20,240 FOR THIS SAME EXAMPLE, THE SAME 1887 01:18:20,240 --> 01:18:23,000 PROJECT, WE CONDUCTED IT WAS 1888 01:18:23,000 --> 01:18:26,760 UNCOMPETITIVE AND NOW SPR 1889 01:18:26,760 --> 01:18:28,800 VERIFIES THAT AND WE HAVE THESE 1890 01:18:28,800 --> 01:18:30,840 TWO INHIBITORS, THEY BIND TO 1891 01:18:30,840 --> 01:18:33,760 DIFFERENT CONFIRMATIONS AND 1892 01:18:33,760 --> 01:18:35,480 ENZYME CONNETICS CAN'T MEASURE 1893 01:18:35,480 --> 01:18:37,600 THE NO SUBSTRATE BOUNDS STATE 1894 01:18:37,600 --> 01:18:38,880 BECAUSE YOU HAVE TO HAVE SOME 1895 01:18:38,880 --> 01:18:40,760 SUBSTRATE TO DO IT AND SO IN A 1896 01:18:40,760 --> 01:18:42,680 WAY, SPR GIVES YOU MORE 1897 01:18:42,680 --> 01:18:43,080 INFORMATION. 1898 01:18:43,080 --> 01:18:45,240 SO HERE IS AN UNCOMPETITIVE 1899 01:18:45,240 --> 01:18:46,920 COMPOUND SO THIS UNCOMPETITIVE 1900 01:18:46,920 --> 01:18:52,960 COMPOUND DOESN'T BIND. 1901 01:18:52,960 --> 01:18:54,200 IT BINDS TO THE SUBSTRATE BOUND 1902 01:18:54,200 --> 01:18:56,240 COMPLEX WHICH IS CONSISTENT WITH 1903 01:18:56,240 --> 01:18:57,520 AN UNCOMPETITIVE MECHANISM AND 1904 01:18:57,520 --> 01:19:00,440 SOMETHING ELSE YOU CAN'T REALLY 1905 01:19:00,440 --> 01:19:03,520 DO BY ASSAY, IT BINDS BETTER TO 1906 01:19:03,520 --> 01:19:04,320 THE PRODUCT. 1907 01:19:04,320 --> 01:19:05,640 IS HE THE WAY THIS COMPOUND 1908 01:19:05,640 --> 01:19:07,800 WORKS IS IT INHIBITS PRODUCT 1909 01:19:07,800 --> 01:19:09,920 RELEASE SO IT BINDS TO THE 1910 01:19:09,920 --> 01:19:13,080 PRODUCT BOUND ENZYME COMPLEX. 1911 01:19:13,080 --> 01:19:15,080 THIS COMPOUND IS NOT NON 1912 01:19:15,080 --> 01:19:16,600 COMPETITIVE AND IT DOESN'T 1913 01:19:16,600 --> 01:19:18,280 REALLY CARE VERY MUCH WHETHER 1914 01:19:18,280 --> 01:19:21,440 THERE'S NUCLEOTIDE OR NO 1915 01:19:21,440 --> 01:19:22,240 NUCLEOTIDE IT BINDS WESTBOUND TO 1916 01:19:22,240 --> 01:19:31,320 BETTER -- ITDOESN'T BOU BIND TON 1917 01:19:31,320 --> 01:19:33,880 THEY'RE SUBSTRATE IT'S NO 1918 01:19:33,880 --> 01:19:36,280 SUBSTRATE ENZYME. 1919 01:19:36,280 --> 01:19:42,200 SO SPR VERIFIES THAT YOU 1920 01:19:42,200 --> 01:19:44,080 DETERMINE BY CONNETICS SO THERE 1921 01:19:44,080 --> 01:19:46,600 ARE MANY INSTRUMENTS FOR SPR AND 1922 01:19:46,600 --> 01:19:49,280 RELATED TECHNOLOGIES, BLI IS A 1923 01:19:49,280 --> 01:19:51,080 RELATED TECHNOLOGY AND THERE ARE 1924 01:19:51,080 --> 01:19:52,360 MANY DIFFERENT FORMATS NOW AND 1925 01:19:52,360 --> 01:19:54,240 MANY OF THESE FORMATS RELY ON A 1926 01:19:54,240 --> 01:20:00,280 FLOW FORMAT. 1927 01:20:00,280 --> 01:20:03,560 BIACORE, SENSIQ AND THERE ARE 1928 01:20:03,560 --> 01:20:04,520 DIFFERENT INSTRUMENTS AND THE 1929 01:20:04,520 --> 01:20:07,720 NOISE THING ABOUT A FLOW BASED 1930 01:20:07,720 --> 01:20:11,680 IS IT MEASURES ACCURATELY. 1931 01:20:11,680 --> 01:20:13,320 THE INSTEAD OF FLOWING MATERIAL 1932 01:20:13,320 --> 01:20:16,400 OVER A SURFACE, YOU PUT YOUR PRE 1933 01:20:16,400 --> 01:20:18,760 TEEN SURFACE IS ON A TIP OF A 1934 01:20:18,760 --> 01:20:21,320 FIBER OPTIC AND YOU DIP IT INTO 1935 01:20:21,320 --> 01:20:23,680 YOUR SOLUTIONS IT RELIES ON 1936 01:20:23,680 --> 01:20:25,040 DIFFUSION FOR BINDING AND 1937 01:20:25,040 --> 01:20:26,120 UNBINDING SO THE MATH IS 1938 01:20:26,120 --> 01:20:27,320 DIFFERENT AND JUST IN MY 1939 01:20:27,320 --> 01:20:30,160 OBSERVATION IT'S NOT AS GOOD AS 1940 01:20:30,160 --> 01:20:31,360 MEASURING OFF RATES. 1941 01:20:31,360 --> 01:20:33,760 SOME OF THE KEY ELEMENTS FOR 1942 01:20:33,760 --> 01:20:35,320 ASSAY DEVELOPMENT. 1943 01:20:35,320 --> 01:20:36,840 THE I AM MOBILIZATION STRATEGY 1944 01:20:36,840 --> 01:20:41,520 AND THE SURFACE CHEMISTRY. 1945 01:20:41,520 --> 01:20:44,960 IF YOU CAN USE A LABEL PROTEIN 1946 01:20:44,960 --> 01:20:47,080 THEN YOU HAVE HAVE MORE ROW 1947 01:20:47,080 --> 01:20:49,680 PRODUCING ABLE ENZYME, MORE 1948 01:20:49,680 --> 01:20:51,400 REPRODUCE ABLE SURFACES AND YOU 1949 01:20:51,400 --> 01:20:53,720 CAN ALSO DO RANDOM IMMUNE 1950 01:20:53,720 --> 01:20:57,520 COUPLING OR RANDOM LYSINE 1951 01:20:57,520 --> 01:20:59,320 COUPLING AND RANDOM LYSINE 1952 01:20:59,320 --> 01:21:04,000 COUPLING, IF YOU ARE DOING 1953 01:21:04,000 --> 01:21:05,200 KINASES, VERY TRICKY IT MAKES ME 1954 01:21:05,200 --> 01:21:07,320 THINK YOU CAN HAVE THIS FOR 1955 01:21:07,320 --> 01:21:11,200 ANIEN TIME THAT HAS LIESINGS IN 1956 01:21:11,200 --> 01:21:11,800 THE SITE. 1957 01:21:11,800 --> 01:21:22,680 IF YOU MOBILIZE AUCOI KINASE, YD 1958 01:21:24,120 --> 01:21:31,400 TO BECAUSE IT'S REACTIVE IT WILL 1959 01:21:31,400 --> 01:21:34,520 STICK TO YOUR CHIP AND IT WON'T 1960 01:21:34,520 --> 01:21:37,480 STICK TO YOUR PROTEIN SO IT'S 1961 01:21:37,480 --> 01:21:38,440 SOMETHING TRICKY YOU HAVE TO 1962 01:21:38,440 --> 01:21:39,840 KEEP IN MIND. 1963 01:21:39,840 --> 01:21:45,000 HOW AM I DOING FOR TIME? 1964 01:21:45,000 --> 01:21:46,960 THE OTHER THING IS YEAH PROTEIN 1965 01:21:46,960 --> 01:21:48,360 STABILITY, ACTIVITY 1966 01:21:48,360 --> 01:21:49,200 CONFIRMATIONAL, FLEXIBILITY ARE 1967 01:21:49,200 --> 01:21:50,720 VERY IMPORTANT SO SOME PRE TEENS 1968 01:21:50,720 --> 01:21:53,320 ARE REALLY UNSTABLE OBJECT THE 1969 01:21:53,320 --> 01:21:54,720 CHIP UNLESS THEY'RE DENSE AND 1970 01:21:54,720 --> 01:21:55,840 THEY'RE CLOSE TO EACH OTHER AND 1971 01:21:55,840 --> 01:21:59,080 THE OPPOSITE CAN ALSO BE TRUE. 1972 01:21:59,080 --> 01:22:02,520 THEY AGGREGATE ON THE SURFACE IS 1973 01:22:02,520 --> 01:22:04,720 THE SURFACE DENSITY HAS TO BE 1974 01:22:04,720 --> 01:22:06,040 SLOW SO THESE ARE THINGS YOU 1975 01:22:06,040 --> 01:22:07,080 NEED TO CONSIDER. 1976 01:22:07,080 --> 01:22:08,520 PROTEIN STABILITY OVER TIME IS 1977 01:22:08,520 --> 01:22:10,600 VERY IMPORTANT AND SURFACE 1978 01:22:10,600 --> 01:22:12,600 ACTIVITY BECAUSE WE'RE GOING TO 1979 01:22:12,600 --> 01:22:13,800 USE, WE'RE GOING TO ASSUME THE 1980 01:22:13,800 --> 01:22:15,160 MASS OF THE PROTEIN ON THE 1981 01:22:15,160 --> 01:22:17,000 SURFACE IS ACTIVE UNLESS PROVEN 1982 01:22:17,000 --> 01:22:20,360 OTHERWISE AND SO YOU ARE STRIKE 1983 01:22:20,360 --> 01:22:22,680 ONLY TREE WILL BE STRONG IF IT'S 1984 01:22:22,680 --> 01:22:23,800 EFFECTIVE SO IF YOU HAVE A 1985 01:22:23,800 --> 01:22:24,960 POSITIVE CONTROL OR SUBSTRATE 1986 01:22:24,960 --> 01:22:27,320 THAT YOU CAN MEASURE THE BINDING 1987 01:22:27,320 --> 01:22:29,200 AND YOU SAY I'M SATURATING MY 1988 01:22:29,200 --> 01:22:31,840 BINDING AT 80% OF WHAT I EXPECT 1989 01:22:31,840 --> 01:22:35,160 SO MY SURFACE 180% ACTIVE IT'S 1990 01:22:35,160 --> 01:22:36,800 FINE JUST KEEP THAT IN MIND WHEN 1991 01:22:36,800 --> 01:22:39,320 YOU ANALYZE YOUR DATA AND THAT 1992 01:22:39,320 --> 01:22:45,520 POSITIVE CONTROL YOU INJECT IT T 1993 01:22:45,520 --> 01:22:47,520 AND SOME USE INTERNAL AUTO MAKE 1994 01:22:47,520 --> 01:22:51,040 GOING ON FOR SEVERAL DAYS. 1995 01:22:51,040 --> 01:22:53,600 SO YOUR PROTEIN SURFACE 1996 01:22:53,600 --> 01:22:54,800 STABILITY TEE IS IMPORTANT AND 1997 01:22:54,800 --> 01:22:56,640 YOU CAN MULTI PLEX. 1998 01:22:56,640 --> 01:23:00,240 MOST OF THESE MODERN INSTRUMENTS 1999 01:23:00,240 --> 01:23:01,160 HAVE -- THEY DIFFER IN HOW 2000 01:23:01,160 --> 01:23:03,960 THEY'RE DESIGNED BUT THIS IS AN 2001 01:23:03,960 --> 01:23:05,680 OLD CHIP AND IT HAS FIVE SPOTS 2002 01:23:05,680 --> 01:23:08,040 PER CHANNEL AND SO EACH OF THESE 2003 01:23:08,040 --> 01:23:11,120 IS A CHANNEL AND SO THAT'S THE 2004 01:23:11,120 --> 01:23:13,040 MICRO FLUID I CAN CHANNEL AND 2005 01:23:13,040 --> 01:23:15,920 THE PLACE THE PROTEIN IS 2006 01:23:15,920 --> 01:23:17,800 IMMOBILIZED IS THIS GREN SPOT ON 2007 01:23:17,800 --> 01:23:21,360 THESE MICRO FLUIDIC CHANNELS AND 2008 01:23:21,360 --> 01:23:23,000 EACH TAKES ONE COMPOUND AT A 2009 01:23:23,000 --> 01:23:25,400 TIME SO ONE AT A TIME AND EACH 2010 01:23:25,400 --> 01:23:28,800 ANNA LIGHT CAN GO OVER ALL FIVE 2011 01:23:28,800 --> 01:23:35,160 SURFACES AT THE SAME TIME SO FOR 2012 01:23:35,160 --> 01:23:39,880 EXAMPLE, ONE THAT HAS YOUR 2013 01:23:39,880 --> 01:23:41,200 PROTEIN OF INTEREST, ONE THAT 2014 01:23:41,200 --> 01:23:42,520 HAS YOUR PROTEIN OF INTEREST 2015 01:23:42,520 --> 01:23:45,960 WITH THE ACTIVE SITE BLOCKED OR 2016 01:23:45,960 --> 01:23:47,800 A DIFFERENT DENSITY OR SOMETHING 2017 01:23:47,800 --> 01:23:50,760 ELSE AND IT CAN BE A RELEVANT 2018 01:23:50,760 --> 01:23:52,560 PROTEIN OR ANNA LOGS OF YOUR 2019 01:23:52,560 --> 01:23:54,640 PROTEINS THAT YOU DON'T WANT TO 2020 01:23:54,640 --> 01:23:55,240 HIT. 2021 01:23:55,240 --> 01:23:57,040 AND REQUIRE DOING A LIPID KINASE 2022 01:23:57,040 --> 01:24:00,520 PROJECT RIGHT NOW WITH NCATS AND 2023 01:24:00,520 --> 01:24:03,000 WE PIT THROW ISOFORMS ON THE 2024 01:24:03,000 --> 01:24:03,440 CHIP AT THE SAME TIME. 2025 01:24:03,440 --> 01:24:05,240 SO THESE ARE ALL THINGS THAT ARE 2026 01:24:05,240 --> 01:24:07,040 EASY TO DO AND YOU BASICALLY GET 2027 01:24:07,040 --> 01:24:09,840 THIS DATA ALL AT THE SAME TIME. 2028 01:24:09,840 --> 01:24:11,880 SO IT'S A VERY DATA-RICH 2029 01:24:11,880 --> 01:24:13,400 TECHNIQUE AND WHEEL COME BACK TO 2030 01:24:13,400 --> 01:24:17,400 THAT IN LEAD OPTIMIZIZATION. 2031 01:24:17,400 --> 01:24:20,600 I WANT TO TALK ABOUT NMR WHICH A 2032 01:24:20,600 --> 01:24:22,120 VERY FLEXIBLE METHODOLOGY THAT 2033 01:24:22,120 --> 01:24:24,000 THERE ARE A LOT OF WAYS TO USE 2034 01:24:24,000 --> 01:24:26,000 IT IN SMALL MOLECULE DISCOVERY 2035 01:24:26,000 --> 01:24:27,280 AND I DON'T KNOW ABOUT YOU GUYS 2036 01:24:27,280 --> 01:24:29,880 BUT I HAVE A HARD TIME 2037 01:24:29,880 --> 01:24:31,400 CONVINCING ANYONE TO DO IT. 2038 01:24:31,400 --> 01:24:34,400 MAYBE I'LL CONVINCE YOU OR YOU 2039 01:24:34,400 --> 01:24:36,200 WILL HAVE MORE BALANCE AS TO 2040 01:24:36,200 --> 01:24:38,720 TAKE BACK TO YOUR LAB TO 2041 01:24:38,720 --> 01:24:40,240 CONVINCE PEOPLE TO DO IT BECAUSE 2042 01:24:40,240 --> 01:24:43,760 THERE ARE SO MANY NMR 2043 01:24:43,760 --> 01:24:45,240 EXPERIENCES YOU CAN DO. 2044 01:24:45,240 --> 01:24:48,640 YOU CAN DO SMALL MOLECULE 2045 01:24:48,640 --> 01:24:50,040 DETECTED NMR. 2046 01:24:50,040 --> 01:24:51,200 DOESN'T USE A LOT OF PROTEIN, 2047 01:24:51,200 --> 01:24:52,040 VERY FAST. 2048 01:24:52,040 --> 01:24:54,160 IT'S PROTEIN NMR BASED 2049 01:24:54,160 --> 01:24:54,600 TECHNIQUES. 2050 01:24:54,600 --> 01:24:56,760 YOU CAN DO PROTEIN NMR WHICH CAN 2051 01:24:56,760 --> 01:24:58,520 BE A LITTLE MORE COSTLY AND IT 2052 01:24:58,520 --> 01:25:00,440 TAKES MORE PROTEIN BUT YOU CAN 2053 01:25:00,440 --> 01:25:02,120 GET BINDING SITE INFORMATION SO 2054 01:25:02,120 --> 01:25:03,360 IT'S INFORMATION RICH TECHNIQUE 2055 01:25:03,360 --> 01:25:04,840 AND THEN THERE ARE OTHER KINDS 2056 01:25:04,840 --> 01:25:07,480 OF FANCY METHODOLOGIES AND I'LL 2057 01:25:07,480 --> 01:25:11,040 SEND YOU TO THIS REVIEW. 2058 01:25:11,040 --> 01:25:12,720 FROM 2008 BUT IT IS STILL 2059 01:25:12,720 --> 01:25:15,480 COMPREHENSIVE AND NOT A LOT OF 2060 01:25:15,480 --> 01:25:19,640 BRAND NEW TECHNIQUES AND WAYS OF 2061 01:25:19,640 --> 01:25:21,160 LOOKING AT THE EVIDENCE OF 2062 01:25:21,160 --> 01:25:23,920 BINDING WHERE THE BINDING SITE 2063 01:25:23,920 --> 01:25:25,440 IS BETWEEN THE MOLECULE AND THE 2064 01:25:25,440 --> 01:25:30,760 PRE TEEN AND SO THESE ARE FANCY 2065 01:25:30,760 --> 01:25:32,200 TECHNIQUES BUT SOME ARE 2066 01:25:32,200 --> 01:25:35,040 ROUTINELY USED SO I'LL TELL YOU 2067 01:25:35,040 --> 01:25:36,000 ABOUT THOSE. 2068 01:25:36,000 --> 01:25:40,040 SO THE MOST COMMON METHODOLOGIES 2069 01:25:40,040 --> 01:25:44,040 TAKE ADVANTAGE OF THE 2070 01:25:44,040 --> 01:25:45,960 INTERACTION BETWEEN THE SMALL 2071 01:25:45,960 --> 01:25:48,920 MOLECULES WHICH HAS A SET OF 2072 01:25:48,920 --> 01:25:51,640 CHARACTERISTICS RATES OF DECAY, 2073 01:25:51,640 --> 01:25:58,400 RATES OF SPIN DOUGH K SPIN DECA. 2074 01:25:58,400 --> 01:26:00,200 THE PROTEIN HAS DIFFERENT ONCE. 2075 01:26:00,200 --> 01:26:05,160 PROTEIN IS BIG, IT MOVES SLOWLY, 2076 01:26:05,160 --> 01:26:06,640 IT'S LAZY AND IT MOVES SLOWLY 2077 01:26:06,640 --> 01:26:09,360 AND ROW LAXES QUICKLY AND SMALL 2078 01:26:09,360 --> 01:26:10,320 MOLECULE IS THE OPPOSITE. 2079 01:26:10,320 --> 01:26:11,720 THEY SPIN QUICKLY THROUGH SPACE 2080 01:26:11,720 --> 01:26:16,000 AND THEY'RE SMALL AND THEY HAVE 2081 01:26:16,000 --> 01:26:16,480 SHARP PEAKS. 2082 01:26:16,480 --> 01:26:19,320 SO WHEN YOU SEE A PROTEIN, A 2083 01:26:19,320 --> 01:26:23,480 SMALL MOLECULE NMR IT LOOKS LIKE 2084 01:26:23,480 --> 01:26:23,680 THIS. 2085 01:26:23,680 --> 01:26:24,520 SHARP PEAKS AND NICE SPLITTINGS 2086 01:26:24,520 --> 01:26:30,680 AND THE PRE TEEN HAS THESE BIG 2087 01:26:30,680 --> 01:26:31,920 BLOBY PEAKS. 2088 01:26:31,920 --> 01:26:42,440 IF YOU BIND -- SO, IF YOU IT IT 2089 01:26:45,920 --> 01:26:47,400 TOUCHES THE PROTEIN IT ADOPTS 2090 01:26:47,400 --> 01:26:48,640 THE PROPERTIES OF THE PROTEIN. 2091 01:26:48,640 --> 01:26:51,000 IT'S LIKE THE NMR VERSION OF A 2092 01:26:51,000 --> 01:26:52,040 FLORENCE IF YOU ARE FAMILIAR 2093 01:26:52,040 --> 01:26:53,640 WITH THAT TECHNIQUE SO YOU HAVE 2094 01:26:53,640 --> 01:26:54,560 SOMETHING SMALL BOUND TO 2095 01:26:54,560 --> 01:26:56,560 SOMETHING BIG IT TAKES ON THE 2096 01:26:56,560 --> 01:26:58,360 MOLECULAR PROPERTIES OF THE BIG 2097 01:26:58,360 --> 01:26:58,720 THING. 2098 01:26:58,720 --> 01:27:01,120 AND SO WE CAN SEE THAT AND HERE 2099 01:27:01,120 --> 01:27:02,480 IS AN EXAMPLE IF YOU HAVE THE 2100 01:27:02,480 --> 01:27:03,920 SMALL MOLECULE BOUND TO A 2101 01:27:03,920 --> 01:27:07,320 PROTEIN SO THE BIG BLOBBY SPEAK 2102 01:27:07,320 --> 01:27:08,520 IS PROTEASOME TEEN AND YOU SEE 2103 01:27:08,520 --> 01:27:11,240 HOW BROADER THESE SMALL MOLECULE 2104 01:27:11,240 --> 01:27:12,880 PEEKS WHEN THERE IS NO PROTEIN 2105 01:27:12,880 --> 01:27:14,440 BOUND BECAUSE SOME OF THAT 2106 01:27:14,440 --> 01:27:15,960 MOLECULE IS BINDING TO THE 2107 01:27:15,960 --> 01:27:18,200 PROTEIN AND TAKING ON THE 2108 01:27:18,200 --> 01:27:19,840 CHARACTERISTICS, THE SPIN 2109 01:27:19,840 --> 01:27:20,920 CHARACTERISTICS OF THE PROTEIN 2110 01:27:20,920 --> 01:27:23,960 AND IF YOU ADD PROTEIN, AND YOU 2111 01:27:23,960 --> 01:27:30,080 DON'T BIND, YOU DON'T SEE ANY 2112 01:27:30,080 --> 01:27:30,320 SHIFTS. 2113 01:27:30,320 --> 01:27:32,680 THIS IS A SATURATION TRANSFER 2114 01:27:32,680 --> 01:27:34,280 DIFFERENCE EXPERIMENT. 2115 01:27:34,280 --> 01:27:36,640 OR WALK LOR EXPERIMENTS, IN ONE 2116 01:27:36,640 --> 01:27:38,920 TECHNIQUE, YOU RADIATE THE 2117 01:27:38,920 --> 01:27:40,320 PROTEIN AND THEY RADIATION IS 2118 01:27:40,320 --> 01:27:44,320 TRANSFERRED TO THE SMALL 2119 01:27:44,320 --> 01:27:45,080 MOLECULE. 2120 01:27:45,080 --> 01:27:47,560 IT CAN TRANSFER IT'S MAG 2121 01:27:47,560 --> 01:27:50,120 NICATION TO THE SMALL MOLECULE 2122 01:27:50,120 --> 01:27:52,680 AND WHEN YOU DO THAT TECHNIQUE, 2123 01:27:52,680 --> 01:27:54,560 YOU CAN SEE ALL KINDS OF 2124 01:27:54,560 --> 01:27:55,600 INTERESTING THINGS, FOR EXAMPLE, 2125 01:27:55,600 --> 01:27:57,400 YOU CAN DO IT IN SUCH A WAY THAT 2126 01:27:57,400 --> 01:27:59,840 YOU SEE THOSE PEAKS THAT ARE 2127 01:27:59,840 --> 01:28:01,000 MOST BROAD END OR MOST EFFECTED 2128 01:28:01,000 --> 01:28:08,280 ARE THOSE THAT ARE MOST BOUND TO 2129 01:28:08,280 --> 01:28:09,240 THE PROTEIN. 2130 01:28:09,240 --> 01:28:12,320 YOU CAN GET A GENERAL IDEA WHICH 2131 01:28:12,320 --> 01:28:23,280 PARTS ARE BURDEN H BURIED AND SY 2132 01:28:24,240 --> 01:28:30,000 BURIED SO ADVANTAGE TO NMR IS 2133 01:28:30,000 --> 01:28:33,880 EVEN JUST PLAIN OLD PROTEIN IF 2134 01:28:33,880 --> 01:28:35,240 YOU DO PROTON YOU CAN SEE THE 2135 01:28:35,240 --> 01:28:36,880 SHIFTS AND THESE LINE 2136 01:28:36,880 --> 01:28:38,000 BROADENNINGS AND THEY'RE FAST 2137 01:28:38,000 --> 01:28:39,880 AND YOU ZOOS A SMALL 2138 01:28:39,880 --> 01:28:42,280 CONCENTRATION OF PROTEIN AND A 2139 01:28:42,280 --> 01:28:43,680 HIGH STICK ONLY TREE OF TOM 2140 01:28:43,680 --> 01:28:46,960 POUND SO IT LOOKS LIKE THE WAY 2141 01:28:46,960 --> 01:28:48,720 YOU DO AN ASSAY WHERE YOU HAVE 2142 01:28:48,720 --> 01:28:49,720 ABOUT 100 TIMES MORE MOLECULE 2143 01:28:49,720 --> 01:28:52,160 THAN PROTEIN AND IF YOU DO IT 2144 01:28:52,160 --> 01:28:54,400 IT'S THE SAME THING HERE AND 2145 01:28:54,400 --> 01:28:58,680 THIS ASSAY RELIES ON FAST 2146 01:28:58,680 --> 01:29:00,720 CONNETICS THAT THEY BIND AND 2147 01:29:00,720 --> 01:29:05,680 ABSORB AND COME OFF IN THE NEXT 2148 01:29:05,680 --> 01:29:06,120 COMPOUND BINDS. 2149 01:29:06,120 --> 01:29:08,080 BECAUSE YOU RELY ON THIS 2150 01:29:08,080 --> 01:29:09,040 AMPLIFICATION OF SIGNAL IT'S 2151 01:29:09,040 --> 01:29:10,400 ONLY GO AHEAD WHEN YOU HAVE FAST 2152 01:29:10,400 --> 01:29:11,320 ON AND OFF RATES. 2153 01:29:11,320 --> 01:29:13,480 SO IT'S THE BEST IN EARLY STAGE 2154 01:29:13,480 --> 01:29:16,240 DISCOVERY BEFORE YOUR COMPOUNDS 2155 01:29:16,240 --> 01:29:20,800 BIND TIGHTLY SO IT'S A VERY NICE 2156 01:29:20,800 --> 01:29:24,000 TECHNIQUE FOR VALIDATING 2157 01:29:24,000 --> 01:29:29,400 SCREENING HITS AND IT'S RATIO OF 2158 01:29:29,400 --> 01:29:30,720 COMPOUND TO PROTEIN AND IT'S 2159 01:29:30,720 --> 01:29:33,600 WHAT YOU GET OUT OF HTS. 2160 01:29:33,600 --> 01:29:36,320 IT'S NOT A 384 WORLD TECHNIQUE 2161 01:29:36,320 --> 01:29:38,160 IT'S STILL INDIVIDUAL COMPOUNDS, 2162 01:29:38,160 --> 01:29:39,560 ONE WAY YOU GET AROUND THE 2163 01:29:39,560 --> 01:29:40,760 SLOWNESS IS IF YOU HAVE 2164 01:29:40,760 --> 01:29:42,040 COMPOUNDS THAT HAVE REALLY 2165 01:29:42,040 --> 01:29:44,760 DISTINCT NMR SPECTRA YOU CAN 2166 01:29:44,760 --> 01:29:46,200 MULTI PLEX THE COMPOUNDS SO YOU 2167 01:29:46,200 --> 01:29:48,480 CAN HAVE 10 COMPOUNDS IN A TUBE 2168 01:29:48,480 --> 01:29:51,040 AT A TIME. 2169 01:29:51,040 --> 01:29:53,520 SO PROTEIN NMR DETECTION IS A 2170 01:29:53,520 --> 01:29:54,920 VERY POWERFUL TECHNIQUE IF YOU 2171 01:29:54,920 --> 01:30:00,200 ARE PROTEIN IS ASSIGNED 2172 01:30:00,200 --> 01:30:02,040 ESPECIALLY IF THE BACKBONE OF 2173 01:30:02,040 --> 01:30:04,320 YOUR PROTEIN IS ASSIGNED SO YOU 2174 01:30:04,320 --> 01:30:12,120 SEE THIS ONE IS A PROTON 2175 01:30:12,120 --> 01:30:15,760 NITROGEN AND YOU CAN DO LARGER 2176 01:30:15,760 --> 01:30:20,440 PROTEINS AND IT'S CALLED AN HMQC 2177 01:30:20,440 --> 01:30:26,720 BUT YOU SEE 2D AXIS EACH DOT 2178 01:30:26,720 --> 01:30:29,080 HERE IS ONE IN THIS EXPERIMENT. 2179 01:30:29,080 --> 01:30:31,680 IT WOULD BE A MENTHOL GROUP SO 2180 01:30:31,680 --> 01:30:37,160 IT'S ONE AM ID BOND SO YOU HAVEA 2181 01:30:37,160 --> 01:30:42,200 SINGLE PEAK ACID AND THESE 2182 01:30:42,200 --> 01:30:44,440 BACKBONE NITROGEN BONDS ARE 2183 01:30:44,440 --> 01:30:45,520 SENSITIVE TO ENVIRONMENT SO IF 2184 01:30:45,520 --> 01:30:47,080 THEY'RE INSIDE THE PROTEIN THEY 2185 01:30:47,080 --> 01:30:48,440 HAVE CHARACTERISTICS SHIFTS IF 2186 01:30:48,440 --> 01:30:51,320 THEY'RE OUTSIDE THE PROTEIN, 2187 01:30:51,320 --> 01:30:52,320 THEY HAVE DIFFERENT SHIFTS AND 2188 01:30:52,320 --> 01:30:54,160 THEY'RE SENSITIVE TO BUFFER AND 2189 01:30:54,160 --> 01:30:58,320 PH BE CAREFUL, SALT BE CAREFUL 2190 01:30:58,320 --> 01:31:01,160 BUT COMPOUND BINDING SO WE DID A 2191 01:31:01,160 --> 01:31:03,080 SCREEN ON THIS SMALL PROTEIN IN 2192 01:31:03,080 --> 01:31:05,720 GRAY AND THIS IS MAKES 2193 01:31:05,720 --> 01:31:06,560 INTERACTIONS SHOWN IN GROWN SO 2194 01:31:06,560 --> 01:31:08,320 WE WERE LOOKING FOR COMPOUNDS 2195 01:31:08,320 --> 01:31:09,960 THAT WOULD COMPETE WITH THIS 2196 01:31:09,960 --> 01:31:11,520 OTHER PROTEIN SUGGESTING THAT 2197 01:31:11,520 --> 01:31:13,960 THEY WOULD SHOW SHIFTS ALONG 2198 01:31:13,960 --> 01:31:16,080 THIS BINDING SITE BUT ALL THE 2199 01:31:16,080 --> 01:31:19,400 COMPOUNDS SHOWED SHIFTS ON THESE 2200 01:31:19,400 --> 01:31:24,080 AROUND THIS BINDING SITE SO 2201 01:31:24,080 --> 01:31:28,960 BACKBONE THAT CORRESPOND SO 2202 01:31:28,960 --> 01:31:30,760 THERE'S A POCKET NEXT TO THE 2203 01:31:30,760 --> 01:31:34,320 MAIN BINDING SITE AND 2204 01:31:34,320 --> 01:31:35,440 INTERESTINGLY, THESE STABILIZE 2205 01:31:35,440 --> 01:31:38,480 SO THEY HELD IT INTO A BINDING 2206 01:31:38,480 --> 01:31:41,320 COM FIRMATION AND SO THAT IS WHY 2207 01:31:41,320 --> 01:31:42,480 I'M SHOWING IT TO YOU HERE 2208 01:31:42,480 --> 01:31:45,240 INSTEAD OF PUBLISHING IT. 2209 01:31:45,240 --> 01:31:46,760 YOU CAN SHOW CLEARLY WELCOME 2210 01:31:46,760 --> 01:31:54,280 WHERE THEIRUSING THIS TECHNIQUEE 2211 01:31:54,280 --> 01:31:56,480 A SHIFT IN THIS RESIDUE IN 2212 01:31:56,480 --> 01:31:56,840 PARTICULAR. 2213 01:31:56,840 --> 01:31:59,080 UPON BINDING. 2214 01:31:59,080 --> 01:32:01,440 EVEN IF YOU DO HAVE THE SPECTRUM 2215 01:32:01,440 --> 01:32:03,600 ASSIGNED, SO YOU SEE ALL THESE 2216 01:32:03,600 --> 01:32:05,480 PEAKS AND YOU DID KNOW WHICH 2217 01:32:05,480 --> 01:32:07,120 AMINO ACID THEY CORRESPOND TO 2218 01:32:07,120 --> 01:32:08,800 YOU CAN GET USEFUL INFORMATION 2219 01:32:08,800 --> 01:32:12,280 SO YOU CAN SAY WELL 10 OF MY 2220 01:32:12,280 --> 01:32:13,800 MOLECULES BIND IN THE SAME PLACE 2221 01:32:13,800 --> 01:32:15,000 OR THE SAME PLACE AS THE 2222 01:32:15,000 --> 01:32:17,760 SUBSTRATE OR THE PROTEIN OR 2223 01:32:17,760 --> 01:32:21,800 SOMETHING SUE CAN STILL GET 2224 01:32:21,800 --> 01:32:23,640 USEFUL INFORMATION. 2225 01:32:23,640 --> 01:32:31,000 SO THE FIRST FRAGMENT WAS SAR BY 2226 01:32:31,000 --> 01:32:34,320 NMR AND IT USED THIS HSQC 2227 01:32:34,320 --> 01:32:35,680 PROTEIN TECHNIQUE BECAUSE IT'S 2228 01:32:35,680 --> 01:32:36,920 SENSITIVE TO THESE ENVIROMENTAL 2229 01:32:36,920 --> 01:32:38,720 CHANGES AND SO THE WAY THIS 2230 01:32:38,720 --> 01:32:40,720 WORKED, SAY YOU HAVE A PROTEIN 2231 01:32:40,720 --> 01:32:42,360 SURFACE, A BINDING SITE WHICH 2232 01:32:42,360 --> 01:32:45,800 TWO SUB POCKETS IN IT AND BY SAR 2233 01:32:45,800 --> 01:32:49,200 BY NMR YOU COULD FIND SHIFTS IN 2234 01:32:49,200 --> 01:32:51,600 THE BINDING POCKET WHEN A 2235 01:32:51,600 --> 01:32:53,120 MOLECULE BOUND THEN YOU OPTIMIZE 2236 01:32:53,120 --> 01:32:54,800 FOR BETTER AND BETTER SHIFTS, 2237 01:32:54,800 --> 01:32:56,840 LARGER SHIFTS, MAYBE MORE 2238 01:32:56,840 --> 01:32:57,440 SHIFTS. 2239 01:32:57,440 --> 01:32:59,160 AND THEN YOU SCREEN FOR A SECOND 2240 01:32:59,160 --> 01:33:02,400 SITE IN THE PRESENCE OF THIS 2241 01:33:02,400 --> 01:33:05,680 OPTIMIZED FIRST SITE FRAGMENT 2242 01:33:05,680 --> 01:33:07,480 AND OPTIMIZE AND THAT FRAGMENT 2243 01:33:07,480 --> 01:33:09,320 AND YOU HAVE TO LINK THESE TWO 2244 01:33:09,320 --> 01:33:13,080 FRAGMENTS IN THE SECOND STEPS 2245 01:33:13,080 --> 01:33:14,720 AND IT'S DRUG DISCOVER ROW BUT 2246 01:33:14,720 --> 01:33:17,560 IT'S POWERFUL TECHNIQUES FOR 2247 01:33:17,560 --> 01:33:18,160 DOING THAT. 2248 01:33:18,160 --> 01:33:20,000 ONE OF THE KEY POINTS THERE IS 2249 01:33:20,000 --> 01:33:22,160 THAT INTERESTINGMENT BASED DRUG 2250 01:33:22,160 --> 01:33:23,840 DISCOVERY USE SMALLER LIBRARIES 2251 01:33:23,840 --> 01:33:25,600 INSTEAD OF USING 100,000 OR A 2252 01:33:25,600 --> 01:33:28,280 MILLION IT USED A THOUSAND TO 2253 01:33:28,280 --> 01:33:29,560 10,000 MOLECULES AND BECAUSE 2254 01:33:29,560 --> 01:33:30,720 THOSE LIBRARIES SIZES ARE 2255 01:33:30,720 --> 01:33:32,400 SMALLER THEY'RE IN THE SAME 2256 01:33:32,400 --> 01:33:34,800 REGIME THAT CAN YOU DO . 2257 01:33:34,800 --> 01:33:36,080 PHYSICAL TECHNIQUES FOR PRIMARY 2258 01:33:36,080 --> 01:33:38,120 SCREENING AND ALSO BECAUSE 2259 01:33:38,120 --> 01:33:39,560 FRAGMENT THOSE LITTLE MOLECULES 2260 01:33:39,560 --> 01:33:46,320 BOUBIND WEAKLY AND ASSAYS CAN BE 2261 01:33:46,320 --> 01:33:47,840 PROBLEMATIC YOU HAVE A LOT OF 2262 01:33:47,840 --> 01:33:49,280 FALSE NEGATIVES AND FALSE 2263 01:33:49,280 --> 01:33:50,240 POSITIVES WHEREAS THESE BINDING 2264 01:33:50,240 --> 01:33:52,520 ASSAYS ARE IN A CONCENTRATION 2265 01:33:52,520 --> 01:33:55,000 REGIME AND MEASURING BINDING 2266 01:33:55,000 --> 01:33:56,560 DIRECTLY AND THEY TEND TO BE 2267 01:33:56,560 --> 01:34:02,400 MUCH LESSER OR PRONE. 2268 01:34:02,400 --> 01:34:05,360 SO A FEW MORE TECHNIQUES, HIGHER 2269 01:34:05,360 --> 01:34:09,160 INFORMATION CONTENT LOWER 2270 01:34:09,160 --> 01:34:11,560 THROUGH PUT TECHNIQUES. 2271 01:34:11,560 --> 01:34:15,000 IT'S COME ALONG IN SPEED PHOTO 2272 01:34:15,000 --> 01:34:16,640 AFFINITY LABELING IS A TECHNIQUE 2273 01:34:16,640 --> 01:34:18,320 THAT YOU MAY SEE PEOPLE USE. 2274 01:34:18,320 --> 01:34:19,960 THE IDEAS THAT YOU TAKE YOUR 2275 01:34:19,960 --> 01:34:22,240 MOLECULE AND YOU ADD SOMETHING 2276 01:34:22,240 --> 01:34:24,320 TO IT THAT WILL BIND TO YOUR 2277 01:34:24,320 --> 01:34:27,240 SMALL MOLECULE, OR YOUR PROTEIN 2278 01:34:27,240 --> 01:34:29,160 AND THE MORE ROW ACTIVE IT IS, 2279 01:34:29,160 --> 01:34:30,600 THE COLLIER HE IS IT BINDS IN 2280 01:34:30,600 --> 01:34:32,000 THE BINDING SITE AND THERE'S 2281 01:34:32,000 --> 01:34:33,960 SEVERAL DIFFERENT REACTIVE 2282 01:34:33,960 --> 01:34:35,680 PROBES THAW MIGHT USE, SOME 2283 01:34:35,680 --> 01:34:38,440 REQUIRE LIGHT, PHOTO A FIN TOE 2284 01:34:38,440 --> 01:34:40,760 LABELING AND OTHERS DO REQUIRE 2285 01:34:40,760 --> 01:34:41,840 LATE THEY'RE JUST ROW ACTIVE AND 2286 01:34:41,840 --> 01:34:44,960 FROM THAT YOU CAN GET AN IDEA 2287 01:34:44,960 --> 01:34:48,400 WHEN YOU THEN CO CHOP UP THE 2288 01:34:48,400 --> 01:34:49,800 PROTEIN YOU CAN FIND OUT WHERE 2289 01:34:49,800 --> 01:34:52,080 THE MOLECULE IS BINDING BASED ON 2290 01:34:52,080 --> 01:34:54,280 WHAT IS ATTACHED TO AND THIS 2291 01:34:54,280 --> 01:34:58,120 GIVES YOU A MODEL FOR HOW THE 2292 01:34:58,120 --> 01:34:58,960 COMPOUND BINDS. 2293 01:34:58,960 --> 01:35:00,400 IT'S NOT USED THAT OFTEN BUT IT 2294 01:35:00,400 --> 01:35:04,000 CAN BE POWERFUL. 2295 01:35:04,000 --> 01:35:06,320 ANOTHER TECHNIQUE CAN BE 2296 01:35:06,320 --> 01:35:07,200 POWERFUL AND I DON'T KNOW HOW 2297 01:35:07,200 --> 01:35:09,200 OFTEN IT'S USED IN INDUSTRY 2298 01:35:09,200 --> 01:35:12,600 BECAUSE IT'S FAIRLY SLOW. 2299 01:35:12,600 --> 01:35:15,240 LOW THROUGH PUT AND IT USES A 2300 01:35:15,240 --> 01:35:16,200 LOT OF MATERIAL. 2301 01:35:16,200 --> 01:35:18,840 IT GIVES YOU VERY NICE DATA. 2302 01:35:18,840 --> 01:35:28,600 THE PRINCIPLES OF ICE OWE NORMAL 2303 01:35:28,600 --> 01:35:29,240 CALIORMETRY IT BINDS HEAT. 2304 01:35:29,240 --> 01:35:32,080 YOU GET ENERGY BY BINDING. 2305 01:35:32,080 --> 01:35:33,560 IT BINDS BECAUSE IT GIVES YOU 2306 01:35:33,560 --> 01:35:35,520 ENERGY AND SO WHEN YOU ADD A 2307 01:35:35,520 --> 01:35:38,800 COMPOUND SO HERE WE PUT THIS IN 2308 01:35:38,800 --> 01:35:40,560 A CAREFULLY TEMPERATURE 2309 01:35:40,560 --> 01:35:41,880 CONTROLLED AND HERE IS THE 2310 01:35:41,880 --> 01:35:43,520 PROTEIN AND WE ADD COMPOUND AND 2311 01:35:43,520 --> 01:35:45,640 AS WE ADD THE COMPOUND IT BIND 2312 01:35:45,640 --> 01:35:47,240 TO THE PROTEIN AND HEATS UP AND 2313 01:35:47,240 --> 01:35:48,840 WE MEASURE THE HEAT AND THE HEAT 2314 01:35:48,840 --> 01:35:54,040 GOES AWAY REALLY QUICKLY AS IT 2315 01:35:54,040 --> 01:35:54,480 RE' CALIBRATION. 2316 01:35:54,480 --> 01:35:56,600 WE GET HEAT AND IT GOES SO AND 2317 01:35:56,600 --> 01:35:58,520 SO UNTIL WE ADD COMPOUND AND WE 2318 01:35:58,520 --> 01:36:01,240 GET NO MORE HEAT BECAUSE THE COM 2319 01:36:01,240 --> 01:36:05,400 UNDER HAS SATURATED THE BINDING 2320 01:36:05,400 --> 01:36:06,000 SITE. 2321 01:36:06,000 --> 01:36:11,160 AND FROM THIS CURVE, YOU CAN GET 2322 01:36:11,160 --> 01:36:13,480 AFFINITY INFORMATION FROM THE 2323 01:36:13,480 --> 01:36:13,840 EC50. 2324 01:36:13,840 --> 01:36:17,000 YOU CAN GET STRIKE ONLY TREE 2325 01:36:17,000 --> 01:36:18,640 INFORMATION BECAUSE IT'S DONE 2326 01:36:18,640 --> 01:36:19,840 QUANTITIES OF COMPOUND AND 2327 01:36:19,840 --> 01:36:24,800 PROTEIN AND YOU GET THE STICK 2328 01:36:24,800 --> 01:36:28,360 ONLY TREE AND WHETHER IT'SEN 2329 01:36:28,360 --> 01:36:29,800 TROPIC DRIVEN SO IF YOU HAVE A 2330 01:36:29,800 --> 01:36:31,320 BIG HEAT THEN IT'S BECAUSE YOU 2331 01:36:31,320 --> 01:36:36,920 ARE GETTING A LOT OF ENTHALPY 2332 01:36:36,920 --> 01:36:38,760 AND IF YOU DON'T GET A HIGH 2333 01:36:38,760 --> 01:36:40,360 SIGNAL, MOST THE BINDING ENERGY 2334 01:36:40,360 --> 01:36:48,480 IS COMING FROM CHANGES IN 2335 01:36:48,480 --> 01:36:53,040 ENTROPY. 2336 01:36:53,040 --> 01:36:55,280 AND IT'S HIGH-THROUGHPUT 2337 01:36:55,280 --> 01:36:56,640 TECHNIQUE A VERY STANDARD 2338 01:36:56,640 --> 01:36:59,240 TECHNIQUE WHEN YOU CAN DO IT BY 2339 01:36:59,240 --> 01:37:00,600 SOAKING YOU CAN ACTUALLY USE IT 2340 01:37:00,600 --> 01:37:02,920 FOR PRIMARY SCREEN AND PEOPLE DO 2341 01:37:02,920 --> 01:37:04,880 THAT FOR FRAGMENT BASED SCREENS 2342 01:37:04,880 --> 01:37:06,680 AND THEY'LL HAVE POOLS OF 10 2343 01:37:06,680 --> 01:37:12,560 MOLECULES AND A SET OF 90 OR 160 2344 01:37:12,560 --> 01:37:13,840 CRYSTALS AND SOAK THE SO MANY 2345 01:37:13,840 --> 01:37:15,160 POUNDS INTO THE CRYSTALS AND SEE 2346 01:37:15,160 --> 01:37:16,480 IF THERE'S DENSITY THERE THAT 2347 01:37:16,480 --> 01:37:19,880 THEY CAN ASSIGN TO THE SMALL 2348 01:37:19,880 --> 01:37:21,280 MOLECULE FRAGMENTS AND 2349 01:37:21,280 --> 01:37:22,920 TECHNOLOGY THAT'S COMING ALONG 2350 01:37:22,920 --> 01:37:25,240 FOR SMALL MOLECULE DISCOVERY IS 2351 01:37:25,240 --> 01:37:28,440 CRYO EM AND I THINK THIS IS 2352 01:37:28,440 --> 01:37:30,240 GOING TO BE AN IMPORTANT 2353 01:37:30,240 --> 01:37:40,640 TECHNOLOGY FOR LEAD O 2354 01:37:41,120 --> 01:37:41,640 OPTIMIZATION. 2355 01:37:41,640 --> 01:37:43,240 IT'S NOT HIGH ENOUGH RESOLUTION 2356 01:37:43,240 --> 01:37:45,320 TO SEE HYDROGEN BONDS BUT YOU 2357 01:37:45,320 --> 01:37:47,280 JUST MODEL THEM ANYWAY, LET'S 2358 01:37:47,280 --> 01:37:48,480 PHASE IT. 2359 01:37:48,480 --> 01:37:50,160 AND LOW RESOLUTION IS REALLY 2360 01:37:50,160 --> 01:37:52,600 WHAT YOU WANT KNOW WHERE IT'S A 2361 01:37:52,600 --> 01:37:54,600 BOUND AND HOW CAN IT MAKE THE 2362 01:37:54,600 --> 01:38:00,040 BOUND BETTER AND IT'S UP TO THAT 2363 01:38:00,040 --> 01:38:00,360 TASK. 2364 01:38:00,360 --> 01:38:03,680 SO WE TALKED ABOUT BINDING 2365 01:38:03,680 --> 01:38:05,680 VALIDATION, MECHANISM OF ACTION 2366 01:38:05,680 --> 01:38:08,520 AND I MENTIONED OFF RATE 2367 01:38:08,520 --> 01:38:08,880 SELECTION. 2368 01:38:08,880 --> 01:38:10,920 AND SO JUST THE POINT IS THAT 2369 01:38:10,920 --> 01:38:11,920 ALL THESE METHODS NEED TO BE 2370 01:38:11,920 --> 01:38:14,120 BALANCED BY SECONDARY SAYS AND 2371 01:38:14,120 --> 01:38:15,560 THAT IS THE GOAL. 2372 01:38:15,560 --> 01:38:16,880 WHATEVER YOU KNOW, THERE'S 2373 01:38:16,880 --> 01:38:20,080 SOMETHING YOU DO KNOW SO USE A 2374 01:38:20,080 --> 01:38:22,080 VARIETY OF TECHNIQUES TO ANSWER 2375 01:38:22,080 --> 01:38:23,720 ALL THE QUESTIONS THAT YOU HAVE 2376 01:38:23,720 --> 01:38:26,720 DO IT EARLY AND DO IT OFTEN, 2377 01:38:26,720 --> 01:38:28,360 DON'T WASTE YOU OR YOUR CHEMIST 2378 01:38:28,360 --> 01:38:30,240 TIME ON ARTIFACTS. 2379 01:38:30,240 --> 01:38:33,760 AND WITH THAT I'LL TURN IT OVER 2380 01:38:33,760 --> 01:38:34,240 TO YOU. 2381 01:38:34,240 --> 01:38:39,480 [APPLAUSE] 2382 01:38:39,480 --> 01:38:41,400 >>THANK YOU SO MUCH FOR AN 2383 01:38:41,400 --> 01:38:42,080 AWESOME TALK. 2384 01:38:42,080 --> 01:38:47,320 QUESTIONS FOR MICHELLE? 2385 01:38:47,320 --> 01:38:47,920 GO AHEAD. 2386 01:38:47,920 --> 01:38:50,840 >>THANK YOU FOR THE NICE TALK. 2387 01:38:50,840 --> 01:38:54,160 I HAVE A QUESTION ABOUT THE DSF 2388 01:38:54,160 --> 01:38:54,520 ASSAYS. 2389 01:38:54,520 --> 01:38:56,560 WE SEE IT TWO PICKS INSTEAD OF 2390 01:38:56,560 --> 01:38:58,600 ONE AND PEM THINK IT'S A MULTI 2391 01:38:58,600 --> 01:38:59,640 DOMAIN SO THAT DO YOU THINK 2392 01:38:59,640 --> 01:39:00,000 ABOUT THAT? 2393 01:39:00,000 --> 01:39:02,400 >>IF YOU HAVE A MULTI DOMAIN 2394 01:39:02,400 --> 01:39:03,760 PROTEIN YOU WILL OFTEN SEE TWO 2395 01:39:03,760 --> 01:39:06,600 PEAKS BECAUSE THEY MELT 2396 01:39:06,600 --> 01:39:07,440 INDEPENDENTLY. 2397 01:39:07,440 --> 01:39:11,360 IF YOU HAVE A SINGLE DOMAIN 2398 01:39:11,360 --> 01:39:14,040 PROTEIN AND YOU ARE GETTING TWO 2399 01:39:14,040 --> 01:39:16,160 PEAKS, I'M NOT SURE. 2400 01:39:16,160 --> 01:39:17,520 IF YOU HAVE SOMETHING SLOWLY 2401 01:39:17,520 --> 01:39:19,920 ININTEREST CONVERTING IT'S 2402 01:39:19,920 --> 01:39:21,400 SIMILAR BUT THAT'S UNUSUAL. 2403 01:39:21,400 --> 01:39:24,360 THANK YOU. 2404 01:39:24,360 --> 01:39:30,800 >>WHAT DO YOU RECOMMEND FOR 2405 01:39:30,800 --> 01:39:33,640 PROTEINS THAT ARE MEMBRANE BOUND 2406 01:39:33,640 --> 01:39:36,320 LIKE GCPR THAT ARE HARD TO 2407 01:39:36,320 --> 01:39:38,640 PURIFY AND SOL YOU SIZE? 2408 01:39:38,640 --> 01:39:42,680 >>WHAT DO I RECOMMEND FOR 2409 01:39:42,680 --> 01:39:43,280 GCPRs? 2410 01:39:43,280 --> 01:39:45,640 PEOPLE HAVE BEEN WORKING ON SPR 2411 01:39:45,640 --> 01:39:47,720 FOR GCPR FOR A LONG TIME AND I 2412 01:39:47,720 --> 01:39:48,920 DON'T KNOW THE LATEST. 2413 01:39:48,920 --> 01:39:51,160 IT'S BEEN DIFFICULT. 2414 01:39:51,160 --> 01:39:53,440 YEAH, USUALLY THE PEOPLE THAT I 2415 01:39:53,440 --> 01:39:59,920 WORK WITH THEY SOL BUELL EYES 2416 01:39:59,920 --> 01:40:04,160 THEM IN DETERGENT AND NANO DISKS 2417 01:40:04,160 --> 01:40:07,920 AND SPINNING SOLID STATE NMR IS 2418 01:40:07,920 --> 01:40:12,960 SOMETHING TO CONSIDER AND LET ME 2419 01:40:12,960 --> 01:40:15,080 THINK WHAT ELSE. 2420 01:40:15,080 --> 01:40:18,440 YOU MAY BE ABLE TO GET 2421 01:40:18,440 --> 01:40:20,480 INFORMATION TOO FROM MASS 2422 01:40:20,480 --> 01:40:22,480 SPECTORMETRY FROM HD EXCHANGE OR 2423 01:40:22,480 --> 01:40:26,880 SOMETHING THAT IS DOU DENATURIND 2424 01:40:26,880 --> 01:40:28,840 YOU CHOP UP THE PROTEIN BUT YOU 2425 01:40:28,840 --> 01:40:31,160 SEE THE ECHO OF WHAT WAS BOUND 2426 01:40:31,160 --> 01:40:33,840 FROM THAT CROSS LINKING OR HD 2427 01:40:33,840 --> 01:40:35,640 EXCHANGE OR PROTECTION FROM 2428 01:40:35,640 --> 01:40:37,920 DIGESTION OR SOMETHING. 2429 01:40:37,920 --> 01:40:40,920 I HAVE TO THINK ABOUT THAT MORE. 2430 01:40:40,920 --> 01:40:44,520 ONE MORE OVER HERE. 2431 01:40:44,520 --> 01:40:49,040 >>I JUST WANTED TO ADD, WORE 2432 01:40:49,040 --> 01:40:52,560 TRYING TO DEVELOP BRETT BASED 2433 01:40:52,560 --> 01:40:54,480 PROBES ON GCPRs SO IT'S 2434 01:40:54,480 --> 01:40:56,160 SOMETHING WE'RE TRYING TO DO 2435 01:40:56,160 --> 01:40:57,440 WITHIN A PEPTIDE BASE PROJECT SO 2436 01:40:57,440 --> 01:41:01,720 WE HAVE A GCPR AND IS EXPRESSING 2437 01:41:01,720 --> 01:41:03,960 NANO LUKE AND THEN THE PEPTIDE 2438 01:41:03,960 --> 01:41:06,760 HAS AN ACCEPTOR. 2439 01:41:06,760 --> 01:41:09,440 >>SO THAT OBJECT INHIBITION 2440 01:41:09,440 --> 01:41:11,520 ANDAL CONTINUE TIVE COMPETITION 2441 01:41:11,520 --> 01:41:11,720 ASSAY. 2442 01:41:11,720 --> 01:41:11,960 >>YES. 2443 01:41:11,960 --> 01:41:12,120 YES. 2444 01:41:12,120 --> 01:41:14,480 >>NOT DIRECT BINDING. 2445 01:41:14,480 --> 01:41:18,520 >>YES. 2446 01:41:18,520 --> 01:41:18,760 CORRECT. 2447 01:41:18,760 --> 01:41:21,920 >>THAT'S USEFUL. 2448 01:41:21,920 --> 01:41:24,200 >>ANYMORE QUESTIONS FOR 2449 01:41:24,200 --> 01:41:24,480 MICHELLE? 2450 01:41:24,480 --> 01:41:26,480 ALL RIGHT, I DON'T SEE ANYMORE 2451 01:41:26,480 --> 01:41:26,680 HANDS. 2452 01:41:26,680 --> 01:41:28,240 THANK YOU SO MUCH, MICHELLE. 2453 01:41:28,240 --> 01:41:28,440 AGAIN. 2454 01:41:28,440 --> 01:41:35,680 [APPLAUSE] 2455 01:41:35,680 --> 01:41:38,840 WE ARE GOING TO TAKE A SHORT 2456 01:41:38,840 --> 01:41:40,160 BREAK AND WE'LL RECONVENE AT 2457 01:41:40,160 --> 01:41:40,440 11:00. 2458 01:41:40,440 --> 01:42:02,160 SEE YOU ALL AT 11:00. 2459 01:42:02,160 --> 01:42:04,920 >>HIS TALK TODAY IS ENTITLED 2460 01:42:04,920 --> 01:42:07,840 "LEAD SELECTION AND 2461 01:42:07,840 --> 01:42:12,800 OPTIMIZIZATION BY MEDICINE 2462 01:42:12,800 --> 01:42:18,840 CHEMISTRY. 2463 01:42:18,840 --> 01:42:21,960 WELCOME. 2464 01:42:21,960 --> 01:42:23,840 >>THANK YOU. 2465 01:42:23,840 --> 01:42:29,640 I'LL TRYING TO FIGURE OUT AND 2466 01:42:29,640 --> 01:42:32,680 AND I WAS TRYING TO BEGIN THIS 2467 01:42:32,680 --> 01:42:36,440 TALK BY ASKING A CHEMIST AND WE 2468 01:42:36,440 --> 01:42:39,760 SAW A WONDERFUL TALKS AND ASSAYS 2469 01:42:39,760 --> 01:42:47,440 AND HOW THEY'RE AND FOR SMALL 2470 01:42:47,440 --> 01:42:50,360 MOLECULES AND WE USE INFORMATION 2471 01:42:50,360 --> 01:42:55,960 FROM THESE ASSAYS AND TO EMILY 2472 01:42:55,960 --> 01:43:04,840 GENERATE STRUCTURE OF AND I 2473 01:43:04,840 --> 01:43:08,840 UNDERSTAND AND WHAT THESE ARE 2474 01:43:08,840 --> 01:43:13,480 AND HOW THEY UTILIZE TO MAKE AND 2475 01:43:13,480 --> 01:43:17,160 HERE IS A A PICTURE AND I WAS 2476 01:43:17,160 --> 01:43:22,120 AND I DIDN'T HAVE FOR AT LEAST A 2477 01:43:22,120 --> 01:43:26,800 YEAR AND WHAT I WANT TO 2478 01:43:26,800 --> 01:43:27,760 ENCOURAGE CHEMIST IS TO 2479 01:43:27,760 --> 01:43:33,240 ESSENTIALLY NOT GO DOWN THIS AND 2480 01:43:33,240 --> 01:43:34,200 WHERE YOU ARE HAVING A 2481 01:43:34,200 --> 01:43:39,280 CONVERSATION WITH THEM AND 2482 01:43:39,280 --> 01:43:45,880 POSITIONING YOURSELF FOR SUCCESS 2483 01:43:45,880 --> 01:43:55,880 AND DEVISING AND MAKING A 2484 01:43:55,880 --> 01:44:06,400 HOLIDAY AND HOW MODERATE WHICH 2485 01:44:31,720 --> 01:44:34,640 WAS DO WE SEE AN EFFECT IN THE 2486 01:44:34,640 --> 01:44:34,960 DISEASE. 2487 01:44:34,960 --> 01:44:40,600 SO I'M GOING TO ACTUALLY 2488 01:44:40,600 --> 01:44:42,600 EXEMPLIFY THESE CONCEPTS BY 2489 01:44:42,600 --> 01:44:44,400 THROW CASE STUDIES THE FIRST 2490 01:44:44,400 --> 01:44:47,200 DEALING WITH KINASE INHIBITORS 2491 01:44:47,200 --> 01:44:49,440 THIS IS CAR SOON STRUCTURE FOR 2492 01:44:49,440 --> 01:44:51,960 KINASE AND IN THIS KINASE, 2493 01:44:51,960 --> 01:44:56,880 THERE'S ATP, THE COFACTOR OF THE 2494 01:44:56,880 --> 01:44:59,840 KINASE WHICH IS THERE'S 2495 01:44:59,840 --> 01:45:02,960 STRUCTURE WHERE YOU NEED LOG 2496 01:45:02,960 --> 01:45:04,880 WHICH HAS A NUMBER OF BETA 2497 01:45:04,880 --> 01:45:06,880 SHEETS AND THEN THERE'S ANOTHER 2498 01:45:06,880 --> 01:45:10,080 DOMAIN WHICH HAS A BUNCH OF 2499 01:45:10,080 --> 01:45:12,600 THOSE CONNECTED BY THIS NARROW 2500 01:45:12,600 --> 01:45:14,840 THREAD OF PEPTIDES WHICH IS 2501 01:45:14,840 --> 01:45:16,200 CALLED THE HINGE REGION AND ALL 2502 01:45:16,200 --> 01:45:18,440 KINASE IS ESSENTIALLY ATP WHICH 2503 01:45:18,440 --> 01:45:24,280 IS THE COFACTOR AND TRANSFER IT 2504 01:45:24,280 --> 01:45:26,960 TO ANOTHER PROTEIN LIPID SO THE 2505 01:45:26,960 --> 01:45:29,840 PORTION BINDS TO THE HINGE 2506 01:45:29,840 --> 01:45:32,720 REGION AND MOST KINASE 2507 01:45:32,720 --> 01:45:34,960 INHIBITORS TROY TO COPY THIS 2508 01:45:34,960 --> 01:45:36,600 CONCEPT SO THEY TRY TO BIND TO 2509 01:45:36,600 --> 01:45:38,000 THE SAME REGION OF THE KINASE. 2510 01:45:38,000 --> 01:45:40,200 THE PROBLEM IS WE HAVE ABOUT 500 2511 01:45:40,200 --> 01:45:45,360 PLUS KINASES AND IN YOU HAVE 2512 01:45:45,360 --> 01:45:48,240 DESIGNING A ATP YOU WILL HAVE CO 2513 01:45:48,240 --> 01:45:50,280 ACTIVITY AGAINST ALL THESE 2514 01:45:50,280 --> 01:45:51,840 KINASES BUT THE FORTUNATE THING 2515 01:45:51,840 --> 01:45:54,160 IS THAT ALTHOUGH THIS DOMAIN OF 2516 01:45:54,160 --> 01:46:00,520 THE KINASE IS OUR CONSERVE THE 2517 01:46:00,520 --> 01:46:02,880 DOMAIN NEARBY AND NOT CON SIDE 2518 01:46:02,880 --> 01:46:03,720 AND YOU CAN EXPLOIT THAT 2519 01:46:03,720 --> 01:46:04,720 DIFFERENCE TO MAKE COMPOUNDS 2520 01:46:04,720 --> 01:46:06,720 WHICH ARE SELECTED AND HERE I'M 2521 01:46:06,720 --> 01:46:09,720 SHOWING YOU TWO VERY FAMOUS 2522 01:46:09,720 --> 01:46:10,760 KINASE INHAD BEEN TOURS AND YOU 2523 01:46:10,760 --> 01:46:12,880 CAN SEE THIS PINK PORTION WHICH 2524 01:46:12,880 --> 01:46:15,160 IS THE REGION BY KINDS TO THE 2525 01:46:15,160 --> 01:46:16,400 REGION AND THERE ARE OTHER 2526 01:46:16,400 --> 01:46:18,080 PORTIONS OF THE COMPOUND THAT 2527 01:46:18,080 --> 01:46:20,360 YOU CAN SEE HOW THEY'RE 2528 01:46:20,360 --> 01:46:21,920 POSITIONED WITHIN THE KINASE AND 2529 01:46:21,920 --> 01:46:23,920 THE CONFIRMATIONS ARE DIFFERENT 2530 01:46:23,920 --> 01:46:26,680 IN THE WAY THEY'RE POSITION AND 2531 01:46:26,680 --> 01:46:29,760 THAT WHAT BUILDS THE 2532 01:46:29,760 --> 01:46:30,040 SELECTIVITY. 2533 01:46:30,040 --> 01:46:32,880 IT'S SHOWN HERE AND THE FIRST 2534 01:46:32,880 --> 01:46:35,600 STUDY I'LL TALK ABOUT ACTUALLY 2535 01:46:35,600 --> 01:46:38,400 BEGAN IN 2005 AND 2008 SO IT'S 2536 01:46:38,400 --> 01:46:40,200 SLIGHTLY DATED BUT I THINK THAT 2537 01:46:40,200 --> 01:46:41,640 THE CONCEPTS THAT I LEARNED ARE 2538 01:46:41,640 --> 01:46:52,160 STILL APPLICABLE TODAY AND AND 2539 01:47:08,720 --> 01:47:16,120 SO THE FIRST CASE STUDY I'LL 2540 01:47:16,120 --> 01:47:19,040 SHOWCASE WHERE WE HAD RECEPTORS 2541 01:47:19,040 --> 01:47:20,800 AND THERE WAS A LOT OF LISTURE 2542 01:47:20,800 --> 01:47:22,160 OUT THERE WHICH MADE SENSE TO GO 2543 01:47:22,160 --> 01:47:24,000 AFTER THIS TARGET AND JUST TO 2544 01:47:24,000 --> 01:47:26,600 QUICKLY GO THROUGH THE SIGNALING 2545 01:47:26,600 --> 01:47:28,600 PATHWAY THIS WAS A KINASE IT'S 2546 01:47:28,600 --> 01:47:31,080 ACTUALLY A DIMER THERE'S AN 2547 01:47:31,080 --> 01:47:37,040 ALPHA SUBUNIT WHEN THE BINDS TO 2548 01:47:37,040 --> 01:47:39,680 THE EXTRA SER YOU LAR DOMAIN AND 2549 01:47:39,680 --> 01:47:41,680 THE DIMER COMES TOGETHER AND DUE 2550 01:47:41,680 --> 01:47:43,320 TO THE CONFORMATIONAL CHANGES 2551 01:47:43,320 --> 01:47:45,240 DUE TO THE BINDING EVENT, WHAT 2552 01:47:45,240 --> 01:47:47,240 HAPPENS IN THE DOMAIN IS THAT 2553 01:47:47,240 --> 01:47:53,200 ONE BETA UNIT RECRUITS ATM AND 2554 01:47:53,200 --> 01:47:57,640 PHOSPHORYLATION THE BETA UNIT 2555 01:47:57,640 --> 01:48:00,160 NEARBY SO THAT LEADS TO 2556 01:48:00,160 --> 01:48:01,240 CONFIRMATION CHANGE AND 2557 01:48:01,240 --> 01:48:02,560 RECRUITMENT OF SEVERAL DIFFERENT 2558 01:48:02,560 --> 01:48:05,640 PLAYERS WHICH ULTIMATELY STARTS 2559 01:48:05,640 --> 01:48:06,800 SIGNALING CASCADES WHICH 2560 01:48:06,800 --> 01:48:09,360 ULTIMATELY ACTUALLY LEAD TO THE 2561 01:48:09,360 --> 01:48:10,880 TURNING ON OF TRANSCRIPTION 2562 01:48:10,880 --> 01:48:11,680 FACTORS WHICH LEAD TO THE 2563 01:48:11,680 --> 01:48:13,640 PRODUCTION OF PROTEINS WHICH 2564 01:48:13,640 --> 01:48:16,320 LEAD TO PROLIFERATION, 2565 01:48:16,320 --> 01:48:16,960 MIGRATION, HALLMARK PHENOTYPES 2566 01:48:16,960 --> 01:48:19,120 OF CANCER. 2567 01:48:19,120 --> 01:48:21,080 NOW, WHILE A LOT OF DATA WITH 2568 01:48:21,080 --> 01:48:24,960 RESPECT TO SILENCING KNOCKING 2569 01:48:24,960 --> 01:48:27,200 OUT THE TARGET INDICATED IT 2570 01:48:27,200 --> 01:48:29,360 WOULD HAVE AN EFFECT ON CELL 2571 01:48:29,360 --> 01:48:30,520 PROLIFERATION AND THE MOST 2572 01:48:30,520 --> 01:48:32,360 IMPORTANT DATA WAS THAT PATIENTS 2573 01:48:32,360 --> 01:48:36,200 WHO WERE BEING TREATED FOR A 2574 01:48:36,200 --> 01:48:38,440 BREAST CANCER WERE BEING TREATED 2575 01:48:38,440 --> 01:48:44,080 WITH HERCEPTIN AND THEY WERE 2576 01:48:44,080 --> 01:48:45,560 DEVELOPING A RESISTANCE AND WHAT 2577 01:48:45,560 --> 01:48:47,760 WAS FOUND IN THE CLINIC WAS THAT 2578 01:48:47,760 --> 01:48:49,760 RESISTANCE WAS DRIVEN BY THIS 2579 01:48:49,760 --> 01:48:51,320 SIGNALING ACCESS AND ONE PROBLEM 2580 01:48:51,320 --> 01:48:54,320 WE REALIZED WAS THAT THE INSULIN 2581 01:48:54,320 --> 01:49:02,040 GROWTH FACTOR ONE REP SCEPTER 2582 01:49:02,040 --> 01:49:04,640 HAS THE INSULIN RECEPTORS AND IF 2583 01:49:04,640 --> 01:49:06,480 WE HAVE COMPOUNDS WE MIGHT HAVE 2584 01:49:06,480 --> 01:49:09,960 A PROBLEM WITH INSULIN SIGNALING 2585 01:49:09,960 --> 01:49:18,360 AND GLUCOSE HOMEOSTASIS, TIM AND 2586 01:49:18,360 --> 01:49:20,160 NATHAN WENT THROUGH A NUMBER OF 2587 01:49:20,160 --> 01:49:21,920 PLATFORMS WHICH CAN BE UTILIZED 2588 01:49:21,920 --> 01:49:27,320 TO DESIGN KINASE INHIBITORS. 2589 01:49:27,320 --> 01:49:31,880 YOU CAN LOOK AT AN ATP AND FOSS 2590 01:49:31,880 --> 01:49:33,160 FIR LAYING OF SUBSTRATE AND 2591 01:49:33,160 --> 01:49:35,160 MONITOR ATP CONCENTRATIONS WHICH 2592 01:49:35,160 --> 01:49:39,800 IS THE COFACTOR AND HAVE 2593 01:49:39,800 --> 01:49:45,440 CELLULAR ASSAYS IN THIS CASE 2594 01:49:45,440 --> 01:49:48,400 ITSELF AND YOU CAN HAVE ASSAYS 2595 01:49:48,400 --> 01:49:50,440 EXPLAINED JUST NOW BY MICHELLE 2596 01:49:50,440 --> 01:49:51,520 AND YOU HAVE ALL THESE DIFFERENT 2597 01:49:51,520 --> 01:49:56,360 THINGS THAT YOU CAN DO 2598 01:49:56,360 --> 01:49:58,120 DOWNSTREAM ASSAYS WHERE YOU LOCK 2599 01:49:58,120 --> 01:50:00,480 AT PHOSPHORYLATION OF DOWNSTREAM 2600 01:50:00,480 --> 01:50:04,400 SUBSTRATES AS WELL AS THE 2601 01:50:04,400 --> 01:50:05,960 TRANSCRIPTION AND PROLIFERATION 2602 01:50:05,960 --> 01:50:06,440 OF CELLS. 2603 01:50:06,440 --> 01:50:07,520 IF YOU HAVE A PLATFORM, YOU CAN 2604 01:50:07,520 --> 01:50:09,000 HAVE THE SAME PLATFORM AND 2605 01:50:09,000 --> 01:50:10,760 EXPAND IT TO OTHER KINASE WHICH 2606 01:50:10,760 --> 01:50:13,000 WILL ALLOW YOU TO THEN FIND OUT 2607 01:50:13,000 --> 01:50:15,760 HOW SELECTIVE YOUR COMPOUNDS 2608 01:50:15,760 --> 01:50:16,320 ARE. 2609 01:50:16,320 --> 01:50:20,240 SO, LET ME QUICKLY DESCRIBE YOU 2610 01:50:20,240 --> 01:50:25,080 THE FIRST SAY WHICH WAS THE 2611 01:50:25,080 --> 01:50:35,600 WORKFORCE ASSAY WE USED AND THIS 2612 01:50:55,840 --> 01:50:57,000 ARTIFICIAL PEPTIDE SUBSTRATE 2613 01:50:57,000 --> 01:50:58,760 THAT'S BIO TIN HERE SO WHEN YOU 2614 01:50:58,760 --> 01:51:00,680 DO THE PHOSPHORYLATION YOU CAN 2615 01:51:00,680 --> 01:51:06,320 ADD AN ANTI BODY A EUROPEAN CRIP 2616 01:51:06,320 --> 01:51:13,560 TATE AND IF YOU ADD ANOTHER 2617 01:51:13,560 --> 01:51:14,840 REALITY WHICH HAS AN ACCEPT OR, 2618 01:51:14,840 --> 01:51:17,600 YOU CAN HAVE A LIGHT TRANSFER 2619 01:51:17,600 --> 01:51:20,160 AND THEN THIS EMITS LIGHT AT A 2620 01:51:20,160 --> 01:51:22,560 DIFFERENT FREQUENCY SO BY 2621 01:51:22,560 --> 01:51:25,160 MEASURING THE FREQUENCY THAT 2622 01:51:25,160 --> 01:51:26,560 THESE LIGHTS, YOU CAN 2623 01:51:26,560 --> 01:51:29,440 ESSENTIALLY FIND OUT HOW MUCH 2624 01:51:29,440 --> 01:51:30,040 PHOSPHORYLATED PEPTIDE YOU HAVE 2625 01:51:30,040 --> 01:51:31,520 AND WHAT YOU WANT TO SEE IS BY 2626 01:51:31,520 --> 01:51:33,680 THE ADDITION OF COMPOUND YOU 2627 01:51:33,680 --> 01:51:37,880 SHOULD SEE A DOSE DEPENDENT 2628 01:51:37,880 --> 01:51:40,120 REDUCTION OF THE SIGNAL WHICH 2629 01:51:40,120 --> 01:51:43,320 TELLS YOU HOW MUCH YOU HAVE OF 2630 01:51:43,320 --> 01:51:44,800 THE SIGNAL YOU HAVE PEPTIDE YOU 2631 01:51:44,800 --> 01:51:45,080 HAVE. 2632 01:51:45,080 --> 01:51:47,240 I'LL BE SHOWING IC50 IN THE 2633 01:51:47,240 --> 01:51:48,920 NUMBER WHICH IS ESSENTIALLY THE 2634 01:51:48,920 --> 01:51:50,440 CONCENTRATION AT I WITH YOU SEE 2635 01:51:50,440 --> 01:51:53,400 50% INHIBITION OF THE SIGNAL AND 2636 01:51:53,400 --> 01:51:55,280 NOW, AS YOU HAVE ALREADY 2637 01:51:55,280 --> 01:51:56,880 REALIZED, THIS IS THE KINASE 2638 01:51:56,880 --> 01:52:00,120 DOMAIN IS INEXTRA CELLULAR SO WE 2639 01:52:00,120 --> 01:52:01,720 HAD ANOTHER ASSAY WHICH ALLOWED 2640 01:52:01,720 --> 01:52:05,120 US TO MEASURE FOSS FOLLOW 2641 01:52:05,120 --> 01:52:06,760 RELATED INCELLS SO IN THIS CASE, 2642 01:52:06,760 --> 01:52:08,680 WE WOULD ACTUALLY TAKE CELLS 2643 01:52:08,680 --> 01:52:12,320 WHICH WERE OVER EXPRESSING THE 2644 01:52:12,320 --> 01:52:15,200 KINASE OF INTEREST AND ADD THE 2645 01:52:15,200 --> 01:52:17,080 COMPOUNDS AND THIS PLATE 2646 01:52:17,080 --> 01:52:21,480 ACTUALLY HAD AN ANTI BODY COATED 2647 01:52:21,480 --> 01:52:23,840 ON THE SURFACE WHICH WAS AN ANTI 2648 01:52:23,840 --> 01:52:28,600 BODY SO THAT WOULD THEN CAP 2649 01:52:28,600 --> 01:52:30,280 AFTER AND WE ADD ANOTHER 2650 01:52:30,280 --> 01:52:32,240 ANTIBODY WHICH RECOGNIZES THE 2651 01:52:32,240 --> 01:52:34,280 FOSS FOR RELATED KINASE AND THIS 2652 01:52:34,280 --> 01:52:37,640 IS CHARGED WITH A CRIP TATE YOU 2653 01:52:37,640 --> 01:52:40,000 ADD RE AGENTS THAT LEAD IT OUT 2654 01:52:40,000 --> 01:52:41,280 AND THAT GENERATES A LIGHT 2655 01:52:41,280 --> 01:52:43,320 SIGNAL WHICH CAN BE MONITORED SO 2656 01:52:43,320 --> 01:52:46,320 THIS LIGHT SIGNAL IS ESSENTIALLY 2657 01:52:46,320 --> 01:52:47,480 REFLECTIVE OF HOW MUCH KINASE 2658 01:52:47,480 --> 01:52:49,040 THAT YOU HAVE AND IF YOU ADD 2659 01:52:49,040 --> 01:52:50,320 COMPOUND, WHAT I WANT TO SEE IS 2660 01:52:50,320 --> 01:52:51,840 THE REDUCTION OF THE SIGNAL SO 2661 01:52:51,840 --> 01:52:53,000 THAT TELLS YOU THAT IF YOU HAVE 2662 01:52:53,000 --> 01:52:56,240 A COMPOUND AND YOU ARE 2663 01:52:56,240 --> 01:52:57,680 INHIBITING THAT SIGNALS WHICH IS 2664 01:52:57,680 --> 01:52:59,320 WHAT YOU WANT. 2665 01:52:59,320 --> 01:53:09,800 SO WE NEED A STARTING WE HAD 2666 01:53:29,080 --> 01:53:30,160 PRONOUNCED CELLS AND THE PROFILE 2667 01:53:30,160 --> 01:53:32,800 WHICH WE WANTED WAS ACTUALLY OF 2668 01:53:32,800 --> 01:53:35,280 GROWTH INHIBITION BUT NOT OF A 2669 01:53:35,280 --> 01:53:35,600 CELL. 2670 01:53:35,600 --> 01:53:37,680 AND WE HAVE SOME OTHER KINASES 2671 01:53:37,680 --> 01:53:40,400 AND HERE THE DATA IS SHOWN IN 2672 01:53:40,400 --> 01:53:43,080 LOG PIC50 WHICH IS THE NEGATIVE 2673 01:53:43,080 --> 01:53:45,480 LOG SO OF IC50 SO IF YOU HAVE A 2674 01:53:45,480 --> 01:53:47,200 NUMBER OF SIX IT'S ONE MICRO 2675 01:53:47,200 --> 01:53:51,040 MOLAR IF IT'S NINE IT'S ONE NANO 2676 01:53:51,040 --> 01:53:53,800 MOLAR AND BEFORE YOU START ANY 2677 01:53:53,800 --> 01:53:56,600 PROGRAM FOR MEDICINAL CHEMIST WE 2678 01:53:56,600 --> 01:53:57,960 CAN LOOK AT THE PRIOR ART SO WE 2679 01:53:57,960 --> 01:54:03,320 WERE LATE IN THE FIELD AND THESE 2680 01:54:03,320 --> 01:54:05,000 COMPOUNDS IN IN LITERATURE SO WE 2681 01:54:05,000 --> 01:54:07,800 COULD ADD THESE TO OUR ASSAYS 2682 01:54:07,800 --> 01:54:09,600 AND MEASURE HOW WE'RE RANKING, 2683 01:54:09,600 --> 01:54:11,240 WHAT IS THE RANK ORDER WITH 2684 01:54:11,240 --> 01:54:13,040 RESPECT TO THESE COMPOUNDS. 2685 01:54:13,040 --> 01:54:15,080 ARE WE IMPROVING OR ARE WE 2686 01:54:15,080 --> 01:54:20,080 BEHIND WHAT'S DO WE NEED TO BE? 2687 01:54:20,080 --> 01:54:26,880 SO THIS IS TO HAVE SI SYNTHESIS 2688 01:54:26,880 --> 01:54:29,200 WHICH CAN BE DIVERSIFIED IN THE 2689 01:54:29,200 --> 01:54:29,360 END. 2690 01:54:29,360 --> 01:54:30,720 I DON'T WANT TO GO THROUGH THE 2691 01:54:30,720 --> 01:54:33,240 WHOLE SYNTHESIS BUT THIS IS FOR 2692 01:54:33,240 --> 01:54:34,720 THE CHEMIST AND WE DISCOVERED 2693 01:54:34,720 --> 01:54:36,040 CHEM TREE WHERE YOU TAKE THIS 2694 01:54:36,040 --> 01:54:38,200 AND YOU ADD ASSAYS, YOU HAVE AN 2695 01:54:38,200 --> 01:54:41,040 INTERNAL CYCLIZATION REACTION 2696 01:54:41,040 --> 01:54:50,000 AND YOU COULD OPEN IT UP IN THIS 2697 01:54:50,000 --> 01:54:51,400 PORTION OF THE MOLECULE AND 2698 01:54:51,400 --> 01:54:53,920 ULTIMATELY YOU COULD MAKE THIS 2699 01:54:53,920 --> 01:54:56,080 SAYINGEL AND GENERATE STRUCTURE 2700 01:54:56,080 --> 01:54:57,960 DIVERSITY AND THIS BOTTOM 2701 01:54:57,960 --> 01:55:01,720 PORTION OF THE MOL YOU MOLECULE. 2702 01:55:01,720 --> 01:55:04,960 IT EVALUATES IN THE ASSAY AND 2703 01:55:04,960 --> 01:55:06,960 SEE WHAT MOVEMENT YOU SEE IN THE 2704 01:55:06,960 --> 01:55:11,400 ACTIVITY WITH RESPECT TO THOSE 2705 01:55:11,400 --> 01:55:21,880 CHANGES AND SO IN YELLOW THE 2706 01:55:23,360 --> 01:55:25,080 ASSAYS I DESCRIBED AND THE PANEL 2707 01:55:25,080 --> 01:55:27,200 WE ALSO HAD ASSAYS WHICH ALLOWED 2708 01:55:27,200 --> 01:55:30,040 US TO MEASURE THE EFFECT IN CELL 2709 01:55:30,040 --> 01:55:31,720 PROLIFERATION AND WE HAD A 2710 01:55:31,720 --> 01:55:32,680 PROTEIN SHIFT ASSAY WHERE WE 2711 01:55:32,680 --> 01:55:36,400 WANTED TO SEE AND BIND TO GO THE 2712 01:55:36,400 --> 01:55:38,080 PROTEIN AND WE WANTED EFFICACY 2713 01:55:38,080 --> 01:55:39,520 IN PD EXPERIMENTS AND WE WERE 2714 01:55:39,520 --> 01:55:41,640 ALSO MONITORING ACTUALLY GROUP 2715 01:55:41,640 --> 01:55:44,240 IN THE LEVELS BECAUSE OF THE 2716 01:55:44,240 --> 01:55:45,400 INSULIN RECEPTORS ACTIVITY AND 2717 01:55:45,400 --> 01:55:47,440 WE HAVE A HOST OF ASSAYS WHICH 2718 01:55:47,440 --> 01:55:49,000 ALLOWED US TO ESSENTIALLY 2719 01:55:49,000 --> 01:55:50,600 MEASURE THE DRUG ABILITY OF THE 2720 01:55:50,600 --> 01:55:54,360 COMPOUND AND IT'S CALL CAN I 2721 01:55:54,360 --> 01:55:56,240 DOSE IT IN A RODENT? 2722 01:55:56,240 --> 01:55:57,560 CAN I SEE SOME EXPOSURE. 2723 01:55:57,560 --> 01:55:58,960 DOES IT REACH THE TUMOR WHICH IS 2724 01:55:58,960 --> 01:56:00,880 THE TARGET OF INTEREST. 2725 01:56:00,880 --> 01:56:02,280 FOR THE INTEREST OF TIME, I'LL 2726 01:56:02,280 --> 01:56:05,720 HAVE ONLY TIME TO JUST TALK 2727 01:56:05,720 --> 01:56:07,600 ABOUT THESE STARRED ITEMS TODAY. 2728 01:56:07,600 --> 01:56:10,320 SO, LET'S LOOK AT A DATA TABLE. 2729 01:56:10,320 --> 01:56:12,920 AND SEE WHAT HAPPENS WHEN WE DIG 2730 01:56:12,920 --> 01:56:14,560 THIS COMPOUND WHICH WAS A 2731 01:56:14,560 --> 01:56:17,680 STARTING POINT AND JUST MODIFY 2732 01:56:17,680 --> 01:56:19,600 THIS SMALL PORTION R1 AND WHAT 2733 01:56:19,600 --> 01:56:22,600 I'M SHOWING YOU HERE IS THE 2734 01:56:22,600 --> 01:56:25,280 ASSAY, IC50 WITH RESPECT TO IGFR 2735 01:56:25,280 --> 01:56:26,640 WHICH IS A TARGET OF INTEREST 2736 01:56:26,640 --> 01:56:30,000 AND YOU HAVE AURORA B AND 2737 01:56:30,000 --> 01:56:31,560 JUNCTION ONE WHICH WERE THE CO 2738 01:56:31,560 --> 01:56:32,800 ACTIVITIES IN THE STARTING 2739 01:56:32,800 --> 01:56:35,840 COMPOUND AND I HAVE THE CELLULAR 2740 01:56:35,840 --> 01:56:37,640 IC15 THE PHOSPHORYLATED ASSAY 2741 01:56:37,640 --> 01:56:45,360 AND WHAT YOU CAN SEE IS THAT BY 2742 01:56:45,360 --> 01:56:46,560 CHANGING THIS GROUP, WE'RE 2743 01:56:46,560 --> 01:56:48,760 SLOWLY ERODING AWAY THE AURORA B 2744 01:56:48,760 --> 01:56:51,400 ACTIVITY IN FACT, IT'S 2745 01:56:51,400 --> 01:56:52,920 COMPLETELY, WE HAVE COMPLETELY 2746 01:56:52,920 --> 01:56:54,080 REMOVED THE ACTIVITY ON THE 2747 01:56:54,080 --> 01:56:58,600 OTHER HAND, THIS BUYS US MORE 2748 01:56:58,600 --> 01:57:09,480 POTENCY WITH RESPECT TO THE IGFR 2749 01:57:17,120 --> 01:57:18,560 NUMBERS HERE WE HAVE SMALLER 2750 01:57:18,560 --> 01:57:18,920 NUMBERS HERE. 2751 01:57:18,920 --> 01:57:20,560 I WANT TO EMPHASIZE THAT 2752 01:57:20,560 --> 01:57:21,880 SOMETIMES ONE TO ONE 2753 01:57:21,880 --> 01:57:23,760 CORRELATIONS DON'T EXIST BUT YOU 2754 01:57:23,760 --> 01:57:26,480 CAN STILL SEE TRENDS AND IT'S 2755 01:57:26,480 --> 01:57:26,840 FINE. 2756 01:57:26,840 --> 01:57:28,200 THE OTHER THING WE ALSO REALIZE 2757 01:57:28,200 --> 01:57:32,520 IS THAT WE CAN ACTUALLY HANG A 2758 01:57:32,520 --> 01:57:36,440 BUNCH OF WATER GROUPS THAT TOLD 2759 01:57:36,440 --> 01:57:39,600 US THIS WAS PROBABLY OUT WITH 2760 01:57:39,600 --> 01:57:40,920 THE SOLVENT FRONT SO WE HAVE 2761 01:57:40,920 --> 01:57:41,280 FLEXIBILITY. 2762 01:57:41,280 --> 01:57:42,920 HOWEVER, THE POINT I WANT TO 2763 01:57:42,920 --> 01:57:47,040 EMPHASIZE IS THAT ROBUST ASSAYS 2764 01:57:47,040 --> 01:57:49,520 ALLOW US TO A ARRIVE AT DATA 2765 01:57:49,520 --> 01:57:50,240 LIKE THIS. 2766 01:57:50,240 --> 01:57:52,560 SO, QUICKLY, I WANT TO SHOW YOU 2767 01:57:52,560 --> 01:57:55,160 SOME IN VIVO DATA AS WELL AND SO 2768 01:57:55,160 --> 01:57:58,520 WHEN A COMPOUND IS DOSED, BY THE 2769 01:57:58,520 --> 01:58:02,760 IV DOSE, YOU OFTEN SEE A CURVE 2770 01:58:02,760 --> 01:58:04,400 LIKE THIS AND THIS IS BECAUSE 2771 01:58:04,400 --> 01:58:07,960 PRPREDOMINANTLY IT REACHES THE 2772 01:58:07,960 --> 01:58:10,440 LIVER WHERE THIS ARE THESE 2773 01:58:10,440 --> 01:58:12,080 WONDERFUL ENZYMES WHICH ACTUALLY 2774 01:58:12,080 --> 01:58:13,800 WILL DO THESE TRANSFORMATIONS 2775 01:58:13,800 --> 01:58:15,960 WHERE THEY TAKE A HYDROGEN AND 2776 01:58:15,960 --> 01:58:17,880 CONVERT IT TO HYDROXY AND THESE 2777 01:58:17,880 --> 01:58:24,680 ARE CALLED P450 IT'S GOOD THAT 2778 01:58:24,680 --> 01:58:25,880 THEY'RE THERE BECAUSE THEY'VE 2779 01:58:25,880 --> 01:58:29,880 HELD PROTECT YOU FROM ZENO 2780 01:58:29,880 --> 01:58:31,680 BIOTICS BUT WE HAVE TO REACH OUR 2781 01:58:31,680 --> 01:58:32,400 TARGET OF INTEREST. 2782 01:58:32,400 --> 01:58:36,200 THIS IS THE FIRST PASS AND OTHER 2783 01:58:36,200 --> 01:58:39,520 ENZYMES WATER AND YOU CAN 2784 01:58:39,520 --> 01:58:40,520 ESSENTIALLY ELIMINATE THESE 2785 01:58:40,520 --> 01:58:41,800 COMPOUNDS AND FOR THE CHEMIST IN 2786 01:58:41,800 --> 01:58:43,440 THE AUDIENCE, THERE'S A NOISE 2787 01:58:43,440 --> 01:58:45,000 MECHANISM FOR YOU TO THINK ABOUT 2788 01:58:45,000 --> 01:58:46,080 HOW THESE WORK. 2789 01:58:46,080 --> 01:58:49,480 SO ANYWAY, IF WE GO BACK TO THE 2790 01:58:49,480 --> 01:58:51,440 IV, DOSING CURVE, WE SEE IS A 2791 01:58:51,440 --> 01:58:52,720 CURVE WHICH LOOKS LIKE THIS AND 2792 01:58:52,720 --> 01:58:54,720 YOU CAN USE PARAMETERS LIKE 2793 01:58:54,720 --> 01:58:56,960 UNDER THE COVER AND THE 2794 01:58:56,960 --> 01:58:57,920 CONCENTRATION OF THE LAST TIME 2795 01:58:57,920 --> 01:58:59,960 POINT TO COMPARE AND CONTRAST 2796 01:58:59,960 --> 01:59:01,400 COMPOUNDS AND IF YOU DOSE A 2797 01:59:01,400 --> 01:59:04,080 COMPOUND ORDERLY, YOU SEE AN AB 2798 01:59:04,080 --> 01:59:07,160 PORTION FACE WHEN IT'S BEING 2799 01:59:07,160 --> 01:59:09,240 ABSORBED INTO THE LIVER AND YOU 2800 01:59:09,240 --> 01:59:11,240 CAN ALSO SEE ELIMINATION PHASE 2801 01:59:11,240 --> 01:59:14,920 YOU CAN USE PARAMETER LIKE THE 2802 01:59:14,920 --> 01:59:16,000 MAXIMUM CONCENTRATION OR THE 2803 01:59:16,000 --> 01:59:20,200 CURVE DIVIDED BY THE DOSE YOU 2804 01:59:20,200 --> 01:59:21,960 CAN USE IT TO COMPARE AND 2805 01:59:21,960 --> 01:59:23,760 CONTRAST AND EXPOSURE YOU ARE 2806 01:59:23,760 --> 01:59:29,520 GETTING THE DIFFERENT AND IN THE 2807 01:59:29,520 --> 01:59:31,800 VERSE RELATIONSHIP AND FROM THE 2808 01:59:31,800 --> 01:59:33,560 CURVE SO IT WAS DATA SO SEE WHAT 2809 01:59:33,560 --> 01:59:36,760 HAPPENS WHEN YOU OUTPUT THIS 2810 01:59:36,760 --> 01:59:41,760 COMPOUND AND MEETING AND WHAT 2811 01:59:41,760 --> 01:59:44,000 HAPPENS WHEN YOU MAKE VERY SMALL 2812 01:59:44,000 --> 01:59:45,800 CHANGES TO THIS PORTION OF THE 2813 01:59:45,800 --> 01:59:48,000 MOLECULE AND IF YOU DID THE MOL 2814 01:59:48,000 --> 01:59:51,160 YOU'LL, WHERE R1 IS HYDROGEN AND 2815 01:59:51,160 --> 01:59:53,200 SO WHAT WE REALIZE IS THAT OUR 2816 01:59:53,200 --> 02:00:01,080 CLEARANCE WAS HIGH AND IF YOU 2817 02:00:01,080 --> 02:00:08,560 MAKE AND YOU SEE AN IMPROVEMENT 2818 02:00:08,560 --> 02:00:10,920 AND CONCENTRATION AT EIGHT HOURS 2819 02:00:10,920 --> 02:00:15,880 AND YOU SEE IMPROVEMENT YOU 2820 02:00:15,880 --> 02:00:26,440 HONOR THOSE AND R2 AND SAME AND 2821 02:00:31,720 --> 02:00:33,120 PAM AR TERES BUT UNFORTUNATELY 2822 02:00:33,120 --> 02:00:43,560 WE'RE LOSING ACTIVITY AND 2823 02:00:43,560 --> 02:00:46,960 ATTENDING THE DATA LIKE THIS AND 2824 02:00:46,960 --> 02:00:49,120 CHEMIST AND YOU CAN USE THESE 2825 02:00:49,120 --> 02:00:51,800 RESULTS AND THEN DICTATE WHICH 2826 02:00:51,800 --> 02:01:01,720 YOU WANT TO KEEP AND ZENO GRAPH 2827 02:01:01,720 --> 02:01:03,400 MODEL AND WHAT WE REALIZE WAS 2828 02:01:03,400 --> 02:01:08,920 THIS HAD A NARROW IN THE CERTAIN 2829 02:01:08,920 --> 02:01:11,520 AND WE SENT BACK AND SAID WAIT A 2830 02:01:11,520 --> 02:01:13,520 SECOND, WE STILL HAVE THE JUNK 2831 02:01:13,520 --> 02:01:14,560 ACTIVITY AND PAPER WE SHOULD 2832 02:01:14,560 --> 02:01:22,160 DIAL IT OUT AND I'M CHANGING AND 2833 02:01:22,160 --> 02:01:30,240 THIS COMPOUND AND HAD THIS FROM 2834 02:01:30,240 --> 02:01:33,080 NITROGEN AND IT AND INTEREST AND 2835 02:01:33,080 --> 02:01:40,040 IT HAD GREAT CELLULAR ACTIVITY 2836 02:01:40,040 --> 02:01:42,320 AND THE JUNK ACTIVITY AND WE'LL 2837 02:01:42,320 --> 02:01:47,120 BE FOUND IS THAT THIS PARTICULAR 2838 02:01:47,120 --> 02:01:57,600 GROUP AND I LOST SELECTIVITY 2839 02:02:01,680 --> 02:02:03,440 AGAINST THE JUNK KINASE SO 2840 02:02:03,440 --> 02:02:05,080 ROBUST REPRODUCIBILITY ASSAYS 2841 02:02:05,080 --> 02:02:08,280 ALLOW YOU TO MAKE THESE SUBTLE 2842 02:02:08,280 --> 02:02:11,560 CONCLUSIONS WITH ROW STRICT TO 2843 02:02:11,560 --> 02:02:14,880 THE RELATIONSHIP. 2844 02:02:14,880 --> 02:02:17,520 THIS WAS ACTUALLY WAS A 2845 02:02:17,520 --> 02:02:18,960 CANDIDATE AND WE DECIDE TODAY 2846 02:02:18,960 --> 02:02:20,760 LOOK AT THE KINASE SELECTIVITY 2847 02:02:20,760 --> 02:02:31,400 AGAINST A BROADER PANEL AN 4% OF 2848 02:02:39,400 --> 02:02:40,680 PATIENTS HAVE A REARRANGEMENT 2849 02:02:40,680 --> 02:02:51,160 WITH PROTEIN AGAIN THE ML4 AND 2850 02:02:59,200 --> 02:03:02,440 WE WERE EVALUATING THE BROAD 2851 02:03:02,440 --> 02:03:03,400 ASSAY PLATFORM SO WE THOUGHT IT 2852 02:03:03,400 --> 02:03:04,800 WAS GREAT AND WE PUSHED THIS 2853 02:03:04,800 --> 02:03:06,600 ACTIVITY AND WENT TO THE 2854 02:03:06,600 --> 02:03:09,320 MANAGEMENT AND THIS IS A GREAT 2855 02:03:09,320 --> 02:03:09,600 CANDIDATE. 2856 02:03:09,600 --> 02:03:12,280 THE FUNNY THING IS THE PROOF OF 2857 02:03:12,280 --> 02:03:17,080 CONCEPT FOR CANCER IS STILL OUT 2858 02:03:17,080 --> 02:03:18,520 THERE AND THE COMPOUNDS FOR 2859 02:03:18,520 --> 02:03:20,440 CLINIC FOR CANCER HAVE NOT SEEN 2860 02:03:20,440 --> 02:03:23,720 THE LIGHT OF DAY. 2861 02:03:23,720 --> 02:03:26,960 BUT, ALK INHIBITORS HAVE. 2862 02:03:26,960 --> 02:03:27,560 UNFORTUNATELY WE WERE TOO 2863 02:03:27,560 --> 02:03:28,560 SUCCESSFUL IN MULTIPLE PROGRAMS 2864 02:03:28,560 --> 02:03:29,960 SO THE MANAGEMENT SAID WE CAN 2865 02:03:29,960 --> 02:03:31,360 JUST TAKE ONE COMPOUND, FORWARD 2866 02:03:31,360 --> 02:03:33,160 IT AND THEY TOOK THE CANDIDATE 2867 02:03:33,160 --> 02:03:37,840 WHICH WAS A DRUG BUT THIS WAS 2868 02:03:37,840 --> 02:03:42,480 SHELVED AND WHEN I JOINED NCATS 2869 02:03:42,480 --> 02:03:43,600 IT GOT APPROVED AND FOUR HAVE 2870 02:03:43,600 --> 02:03:45,640 BEEN APPROVED SO THIS IS MY I 2871 02:03:45,640 --> 02:03:49,280 COULD HAVE BEEN THERE. 2872 02:03:49,280 --> 02:03:52,680 SO YOU KNOW, SO IN THE END, YOU 2873 02:03:52,680 --> 02:03:55,400 HAVE A REPORT CARD OF THE 2874 02:03:55,400 --> 02:04:05,960 COMPOUND AND ALL THE ACTIVITIES 2875 02:04:11,200 --> 02:04:13,880 AND TALK TO YOUR TEAMMATES WHO 2876 02:04:13,880 --> 02:04:15,480 GENERATE THIS DATA AND HAVE A 2877 02:04:15,480 --> 02:04:17,480 GRASP OF HOW THIS DATA HAS BEEN 2878 02:04:17,480 --> 02:04:20,120 GENERATED AND WHAT IT MEANS AND 2879 02:04:20,120 --> 02:04:22,840 SHIFT GEARS TO THE NEXT KINASE 2880 02:04:22,840 --> 02:04:23,760 HIN HIB TO BE PROJECT WHERE WE 2881 02:04:23,760 --> 02:04:27,480 HAD A COMPOUND FOR DUCHENE 2882 02:04:27,480 --> 02:04:37,920 MUSCULAR DYSTROPHY SO IS IT 2883 02:04:37,920 --> 02:04:39,600 EFFECTS 1/5,000 BOYS AND I'M 2884 02:04:39,600 --> 02:04:42,400 SHOWING YOU THIS CARTOON WHERE 2885 02:04:42,400 --> 02:04:48,400 I'M SHOWING YOU THIS SARCOLAMA. 2886 02:04:48,400 --> 02:04:50,680 IN THE UNITS OF MUSCLE CELLS 2887 02:04:50,680 --> 02:04:53,960 THEY ACTUALLY ALIGN AND FORM 2888 02:04:53,960 --> 02:04:55,440 THESE TUBES AND SO THE PLASMA 2889 02:04:55,440 --> 02:04:58,560 MEMBRANE WHICH IS CALLED IS 2890 02:04:58,560 --> 02:05:04,800 ALMOST LIKE A CONTINUOUS SORT OF 2891 02:05:04,800 --> 02:05:06,120 STRUCTURE WHERE WE HAVE ALL 2892 02:05:06,120 --> 02:05:10,400 THESE DIFFERENT STRUCTURE 2893 02:05:10,400 --> 02:05:11,520 PROTEINS, ALL THESE DIFFERENT 2894 02:05:11,520 --> 02:05:12,960 STRUCTURE PROTEINS WHICH REALLY 2895 02:05:12,960 --> 02:05:15,280 GIVE A LOT OF STRUCTURE AND 2896 02:05:15,280 --> 02:05:16,760 RIGIDITY TO THE MUSCLE AND 2897 02:05:16,760 --> 02:05:23,440 STRENGTH AND WHAT HAPPENS IN DUE 2898 02:05:23,440 --> 02:05:25,440 LANE, IN THIS PRE TEEN, THIS 2899 02:05:25,440 --> 02:05:27,600 STRUCTURE OF PROTEIN IS ABSENT 2900 02:05:27,600 --> 02:05:30,800 AND THAT MAKES THIS THE MUSCLES 2901 02:05:30,800 --> 02:05:32,920 EXTREMELY WEAK IN THESE YOUNG 2902 02:05:32,920 --> 02:05:35,080 MEN AND WHAT OUR COLLABORATORS 2903 02:05:35,080 --> 02:05:37,440 THOUGHT, WELL, WHAT IF WE UP 2904 02:05:37,440 --> 02:05:38,280 REGULATED THE PRODUCTION OF 2905 02:05:38,280 --> 02:05:40,280 ANOTHER STRUCTURE PROTEIN AND 2906 02:05:40,280 --> 02:05:43,120 MAYBE WE CAN COM PEN TATE FOR 2907 02:05:43,120 --> 02:05:46,400 THE LOSS AND THE ASSAYS WAS 2908 02:05:46,400 --> 02:05:47,800 DESIGNED IN THE WAY THAT WE 2909 02:05:47,800 --> 02:05:51,560 LOOKED AT THAT ALPHA WHICH IS A 2910 02:05:51,560 --> 02:05:52,760 STRUCTURAL PROTEIN AND WE TOOK 2911 02:05:52,760 --> 02:05:57,600 ONE OF THE ALLELES OUT FROM THE 2912 02:05:57,600 --> 02:06:03,640 ALPHA 7 INTRA GRIN PROTEIN AND 2913 02:06:03,640 --> 02:06:05,320 AND IF THAT PROTEIN IS 2914 02:06:05,320 --> 02:06:06,560 EXPRESSED, YOU SEE AN EXPRESSION 2915 02:06:06,560 --> 02:06:09,000 OF THIS REPORTER AND YOU CAN ADD 2916 02:06:09,000 --> 02:06:12,040 THE SUBSTRATE AND GET A SIGNAL 2917 02:06:12,040 --> 02:06:14,520 SO THE LIGHT SIGNAL ESSENTIALLY 2918 02:06:14,520 --> 02:06:15,360 REFLECTS HOW MUCH PROTEIN THAT 2919 02:06:15,360 --> 02:06:18,000 YOU HAVE AND WHEN WE DID 2920 02:06:18,000 --> 02:06:19,200 REPURPOSE AND SCREEN WE 2921 02:06:19,200 --> 02:06:21,720 DISCOVERED THIS COMPOUND THE 2922 02:06:21,720 --> 02:06:23,480 CDK2 INHIBITOR WHICH WAS ABLE TO 2923 02:06:23,480 --> 02:06:25,080 UP REGULATE THE PROTECTION OF 2924 02:06:25,080 --> 02:06:32,600 THIS PROTEIN AND THIS PICTURE OF 2925 02:06:32,600 --> 02:06:35,560 THE -- WHAT IT IS, I'M SORRY, 2926 02:06:35,560 --> 02:06:38,680 MIO TUBES, YOU CAN SEE IN THE 2927 02:06:38,680 --> 02:06:41,800 MIO TUBES THERE'S A GRO GREEN GL 2928 02:06:41,800 --> 02:06:43,320 TELLING YOU THIS COMPOUND IS 2929 02:06:43,320 --> 02:06:44,920 ABLE TO UP REGULATE THE PROCESS 2930 02:06:44,920 --> 02:06:46,360 OF THE PROTEIN. 2931 02:06:46,360 --> 02:06:47,640 THAT WAS GREAT SO THEN THE 2932 02:06:47,640 --> 02:06:50,960 QUESTION IS, WELL, IT'S A CDK2 2933 02:06:50,960 --> 02:06:52,240 INHIBITOR KNOWN IN LITERATURE SO 2934 02:06:52,240 --> 02:06:53,800 THAT PROBABLY IS OUR TARGET BUZZ 2935 02:06:53,800 --> 02:06:57,840 IT'S A PHENOTYPIC ASSAY SO WE 2936 02:06:57,840 --> 02:06:59,240 LET LET'S FIND OUT THE KINASE 2937 02:06:59,240 --> 02:07:01,640 THIS COMPOUND IS ENGAGING WITH 2938 02:07:01,640 --> 02:07:03,240 IN THESE MIO TUBES AND SO I HAVE 2939 02:07:03,240 --> 02:07:05,040 TO INTRODUCE YOU TO THIS 2940 02:07:05,040 --> 02:07:12,320 TECHNOLOGY PLATFORM HERE AND 2941 02:07:12,320 --> 02:07:15,000 WHERE YOU HAVE THESE PROBLEMS 2942 02:07:15,000 --> 02:07:19,200 THAT LOOK LIKE ATP AND WHILE 2943 02:07:19,200 --> 02:07:21,960 TENDS HERE YOU HAVE A CARBON 2944 02:07:21,960 --> 02:07:24,200 HE'LL GROUP WHICH ACTUALLY IS 2945 02:07:24,200 --> 02:07:27,200 REACTIVE AND IF YOU ARE PAYING 2946 02:07:27,200 --> 02:07:29,240 ATTENTION, TO THE TOP BEFORE, 2947 02:07:29,240 --> 02:07:32,920 THERE ARE CONSERVED LICE EVENS 2948 02:07:32,920 --> 02:07:34,880 IN KINASE AT THE OUTER PARK ET 2949 02:07:34,880 --> 02:07:38,320 AND THESE CONSERVE ACTUALLY CAN 2950 02:07:38,320 --> 02:07:39,400 REACT WITH PROBLEMS LIKE THIS 2951 02:07:39,400 --> 02:07:41,440 AND THIS PROBE IS CHARGED WITH 2952 02:07:41,440 --> 02:07:43,200 BIO TIN SO WHEN THIS PROBE 2953 02:07:43,200 --> 02:07:47,360 INTERACTS WITH KINASES IN CELLS, 2954 02:07:47,360 --> 02:07:52,440 AND THIS THE LIE SEEN IS 2955 02:07:52,440 --> 02:08:02,960 ATTACHED TO THIS AND THEN CAN 2956 02:08:13,960 --> 02:08:16,400 YOU ADD A PRO TEE ACE AND CHOP 2957 02:08:16,400 --> 02:08:19,520 OFF THE KINASE AND YOU HAVE A 2958 02:08:19,520 --> 02:08:25,560 UNIQUE PEPTIDE SIGNATURE FROM 2959 02:08:25,560 --> 02:08:36,080 EACH FROM BINDING SO WHEN YOU DO 2960 02:08:38,480 --> 02:08:42,920 IT, YOU WILL ACTUALLY LOSE THAT 2961 02:08:42,920 --> 02:08:48,480 SIGNAL FROM THE LCMS EXPERIMENT. 2962 02:08:48,480 --> 02:08:56,120 SO, LET'S SEE WHAT HAPPENS WHERE 2963 02:08:56,120 --> 02:08:58,240 IT'S SHOWING THE REGULATION OF 2964 02:08:58,240 --> 02:09:00,160 THE ALPHABET TA PROTEIN SO I 2965 02:09:00,160 --> 02:09:01,360 DON'T WANT TO GO THROUGH THIS 2966 02:09:01,360 --> 02:09:03,000 BUT I WANT TO SHOW YOU HOW THE 2967 02:09:03,000 --> 02:09:04,560 DATA LOOKS SO YOU HAVE ALL THESE 2968 02:09:04,560 --> 02:09:06,600 DIFFERENT KINASES AND THESE PEP 2969 02:09:06,600 --> 02:09:08,160 SIDES AND WE HAVE THREE 2970 02:09:08,160 --> 02:09:09,400 DIFFERENT CONCENTRATIONS SO 2971 02:09:09,400 --> 02:09:11,000 GREEN IS WE DON'T SEE A BINDING 2972 02:09:11,000 --> 02:09:13,360 EVENT AND RED WE SEE A GOOD 2973 02:09:13,360 --> 02:09:16,520 BINDING EVENT AND IF WE FOCUS ON 2974 02:09:16,520 --> 02:09:17,920 THE KINASE WHERE WE SHOW 2975 02:09:17,920 --> 02:09:21,320 ACTIVITY OR BINDING OF THIS 2976 02:09:21,320 --> 02:09:23,760 PROBE WE SEE CDK2 AND BINDING IN 2977 02:09:23,760 --> 02:09:25,160 ALL THREE DIFFERENT 2978 02:09:25,160 --> 02:09:27,760 CONCENTRATIONS AND WE SEE CDK4, 2979 02:09:27,760 --> 02:09:30,360 5 WHICH IS EXPECTED AND WE 2980 02:09:30,360 --> 02:09:32,280 DISCOVER SOME OTHER KINASES 2981 02:09:32,280 --> 02:09:34,360 WHERE WE SEE MUCH BETTER 2982 02:09:34,360 --> 02:09:38,840 BINDINGS THAN CDK2 WHICH IS THE 2983 02:09:38,840 --> 02:09:40,000 ORIGINAL TARGET. 2984 02:09:40,000 --> 02:09:41,240 THAT WAS VERY SURPRISING. 2985 02:09:41,240 --> 02:09:42,720 WE SAID WELL MAYBE, THE ACTIVITY 2986 02:09:42,720 --> 02:09:44,640 OF THIS COMPOUND AGAINST THESE 2987 02:09:44,640 --> 02:09:46,440 KINASES IS ALREADY KNOWN AND 2988 02:09:46,440 --> 02:09:48,360 SURE ENOUGH, REACTION BIOLOGY 2989 02:09:48,360 --> 02:09:50,240 WHICH IS A PLATFORM WHICH 2990 02:09:50,240 --> 02:09:54,880 UTILIZES THIS SORT OF TECHNOLOGY 2991 02:09:54,880 --> 02:09:56,800 WHERE THEY HAVE ARTIFICIAL 2992 02:09:56,800 --> 02:10:00,520 SUBSTRATES WHICH IS FOSS FOR 2993 02:10:00,520 --> 02:10:03,040 LATED BY A RADIOACTIVE PHOSPHATE 2994 02:10:03,040 --> 02:10:04,920 AND PUBLISH THIS WONDERFUL PAPER 2995 02:10:04,920 --> 02:10:06,800 WHERE THEY TOOK COMPOUNDS AND 2996 02:10:06,800 --> 02:10:08,560 THEY DID PROFILING AND PUBLISHED 2997 02:10:08,560 --> 02:10:10,080 THIS LITERATURE AND WHEN WE 2998 02:10:10,080 --> 02:10:13,880 LOOKED AT OUR COMPOUNDS, SU9516 2999 02:10:13,880 --> 02:10:16,240 WE SAW THAT AS PREDICTED, IT WAS 3000 02:10:16,240 --> 02:10:18,120 A GREAT INHIBITOR OF THIS 3001 02:10:18,120 --> 02:10:21,320 FUNCTIONAL ACTIVITY AGAINST CDK5 3002 02:10:21,320 --> 02:10:23,240 BUT IT WAS DEAD AGAINST THE TWO 3003 02:10:23,240 --> 02:10:25,640 KINASES THAT WE HAD DISCOVERED 3004 02:10:25,640 --> 02:10:27,600 IN THE MIO TUBE EXPERIMENT SO 3005 02:10:27,600 --> 02:10:29,160 THAT TELLS YOU THAT DEPENDING ON 3006 02:10:29,160 --> 02:10:31,320 THE PLATFORM YOU HAVE, YOU CAN 3007 02:10:31,320 --> 02:10:32,880 HAVE VERY DIFFERENT RESULTS AND 3008 02:10:32,880 --> 02:10:35,000 OBVIOUSLY THE QUESTION IS, WHAT 3009 02:10:35,000 --> 02:10:36,400 IS THE TRUTH? 3010 02:10:36,400 --> 02:10:38,760 THE BIOCHEMICAL OR THIS ONE AND 3011 02:10:38,760 --> 02:10:40,960 SO, YOU KNOW, JUST WANT TO HIT 3012 02:10:40,960 --> 02:10:51,480 HOME THIS CONCEPT WHERE YOU TAKE 3013 02:11:20,120 --> 02:11:21,360 CELLS OF COMPOUNDS WITHOUT 3014 02:11:21,360 --> 02:11:23,160 COMPOUNDS AND TAKE THE LICE IT'S 3015 02:11:23,160 --> 02:11:26,000 A AND INCUBATE WITH THESE BEADS 3016 02:11:26,000 --> 02:11:27,160 AND THEY WILL BIND TO THE KINASE 3017 02:11:27,160 --> 02:11:29,040 AND PULL THEM OUT AND YOU CAN DO 3018 02:11:29,040 --> 02:11:30,240 IT AGAIN AND IF YOU HAVE 3019 02:11:30,240 --> 02:11:31,880 COMPETING COMPOUNDS YOU WON'T 3020 02:11:31,880 --> 02:11:33,280 SEE THAT BINDING SO THIS IS A 3021 02:11:33,280 --> 02:11:34,600 GREAT PAPER AND CAN YOU READ AND 3022 02:11:34,600 --> 02:11:36,640 HERE TOO, THEY FOUND DIFFERENCES 3023 02:11:36,640 --> 02:11:38,560 BETWEEN BIOCHEMICAL AND CELLULAR 3024 02:11:38,560 --> 02:11:40,760 TARGETING ENGAGEMENT. 3025 02:11:40,760 --> 02:11:41,840 ALL RIGHT. 3026 02:11:41,840 --> 02:11:45,000 LAST CASE STUDY, I'LL DISCUSS A 3027 02:11:45,000 --> 02:11:52,400 RARE DISEASE AND IT'S GOUSHER 3028 02:11:52,400 --> 02:11:54,600 DISEASE, THIS IS A RARE 3029 02:11:54,600 --> 02:12:01,840 RECESSIVE DISORDER AND DISEASE 3030 02:12:01,840 --> 02:12:12,320 AND BECAUSE OF THE LACK OF 3031 02:12:29,680 --> 02:12:31,400 ACTIVITY BECAUSE OF THIS ENZYME 3032 02:12:31,400 --> 02:12:34,600 YOU SEE ACCUMULATIONS OF THESE 3033 02:12:34,600 --> 02:12:37,280 SUBSTRATES WITHIN THE LICE OWE 3034 02:12:37,280 --> 02:12:38,240 SOME THAT'S WHY IT'S THE 3035 02:12:38,240 --> 02:12:40,040 DISORDER AND THERE ARE THREE 3036 02:12:40,040 --> 02:12:41,640 FORMS OF THIS DISEASE I'LL FOCUS 3037 02:12:41,640 --> 02:12:44,560 ON EXTRA THE TYPE ONE AND IT HAS 3038 02:12:44,560 --> 02:12:45,920 ESSENTIALLY MOSTLY MAN FESS 3039 02:12:45,920 --> 02:12:50,480 INDICATIONS IN THE VISCERA AND 3040 02:12:50,480 --> 02:12:51,720 THEY HAVE LARGE SPLEENS AND 3041 02:12:51,720 --> 02:12:55,360 LIVES AND THEN THERE ARE TWO 3042 02:12:55,360 --> 02:12:59,600 UNFORTUNATE TYPES WHICH HAVE A 3043 02:12:59,600 --> 02:13:08,360 PATHIC COMPONENT AND ONE OF THE 3044 02:13:08,360 --> 02:13:10,840 GENOTYPES RENDERS IT UNSTABLE. 3045 02:13:10,840 --> 02:13:12,720 IF YOU HAVE THE MOST COMMON 3046 02:13:12,720 --> 02:13:15,880 SUBTYPE OF THIS HYDRO LAYS, IF 3047 02:13:15,880 --> 02:13:19,960 IT MAKES IT TO THE LICE OSEM IT 3048 02:13:19,960 --> 02:13:22,600 HAS CATALYTIC ACTIVITY BUT THE 3049 02:13:22,600 --> 02:13:25,320 PORTING DOESN'T FOLD PROPERLY SO 3050 02:13:25,320 --> 02:13:26,640 IT'S TAGGED FOR DEGRADATION SO 3051 02:13:26,640 --> 02:13:29,320 THE PROBLEM IS YOU HAVE LESS 3052 02:13:29,320 --> 02:13:29,720 PROTEIN. 3053 02:13:29,720 --> 02:13:31,560 SO, THE QUESTION IS HOW DO YOU 3054 02:13:31,560 --> 02:13:34,880 THEN FIND COMPOUNDS WHICH CAN BE 3055 02:13:34,880 --> 02:13:38,040 THERAPEUTIC AND OUR GOAL WAS TO 3056 02:13:38,040 --> 02:13:40,240 ACTUALLY DEVELOP SMALL GROUPS 3057 02:13:40,240 --> 02:13:42,160 AND THIS WAS ALSO A CONCEPT 3058 02:13:42,160 --> 02:13:43,680 EXPLAINED IN THE PREVIOUS TALK, 3059 02:13:43,680 --> 02:13:46,200 YOU CAN SOMETIMES FIND COMPOUNDS 3060 02:13:46,200 --> 02:13:49,160 WHICH BIND TO PROTEINS AND MAKE 3061 02:13:49,160 --> 02:13:51,200 THEM STABLE AND PREVENT THEM 3062 02:13:51,200 --> 02:13:53,560 FROM DEGRADATION SO THAT WAS OUR 3063 02:13:53,560 --> 02:13:53,920 GOAL. 3064 02:13:53,920 --> 02:13:56,280 THE QUESTION WAS LET'S DESIGN . 3065 02:13:56,280 --> 02:13:56,640 CHEMICAL ASSAY. 3066 02:13:56,640 --> 02:14:00,080 WE CAN OBVIOUSLY HAVE RECOMMON 3067 02:14:00,080 --> 02:14:01,360 PROTEIN AND HERE IS WHAT THE 3068 02:14:01,360 --> 02:14:04,440 PROTEIN DOES, IT FORMS A PART OF 3069 02:14:04,440 --> 02:14:06,960 THE MEMBRANE AND IT ACTUALLY 3070 02:14:06,960 --> 02:14:09,400 INTERACTS WITH CO FACTORS, NOT 3071 02:14:09,400 --> 02:14:12,640 CO FACTORS, OTHER PROTEINS WHICH 3072 02:14:12,640 --> 02:14:16,200 HELP ACTUALLY ORIENT THE 3073 02:14:16,200 --> 02:14:20,200 SUBSTRATE IN A PROPER WAY SO IT 3074 02:14:20,200 --> 02:14:20,920 CAN CHOP IT. 3075 02:14:20,920 --> 02:14:26,800 THE PROBLEM WITH THE N37S IT WAS 3076 02:14:26,800 --> 02:14:28,320 HARD TO MAKE IT BECAUSE IT WAS 3077 02:14:28,320 --> 02:14:32,560 UNSTABLE SO WE WENT TO A 3078 02:14:32,560 --> 02:14:35,040 CLINICIAN AND BECAUSE THE 3079 02:14:35,040 --> 02:14:38,200 PATIENTS HAVE ENLARGED SPLEEN, 3080 02:14:38,200 --> 02:14:39,400 SHE HAD SPLEEN SAMPLES FROM THE 3081 02:14:39,400 --> 02:14:43,040 PATIENTS IN HER FREEZER SO WE 3082 02:14:43,040 --> 02:14:44,720 TOOK THOSE SPLEENS AND PUT THEM 3083 02:14:44,720 --> 02:14:45,800 IN A GRINDER AND WE HAD A SOUP 3084 02:14:45,800 --> 02:14:49,240 THAT HAD THIS ENZYME AND THAT 3085 02:14:49,240 --> 02:14:49,960 ENZYME PRESENT IN THE SPLEEN, 3086 02:14:49,960 --> 02:14:53,400 WAS ABLE TO ACTUALLY HAD CAT A 3087 02:14:53,400 --> 02:14:54,080 LAD I CAN FUNCTION AND THE 3088 02:14:54,080 --> 02:14:55,640 QUESTION IS WHAT DO WE HAVE FOR 3089 02:14:55,640 --> 02:14:56,080 SUBSTRATE? 3090 02:14:56,080 --> 02:14:58,480 WE HAVE THE TYPICAL ACTUAL 3091 02:14:58,480 --> 02:15:00,720 SUBSTRATE AND MONITORING THE 3092 02:15:00,720 --> 02:15:02,840 HYDROLYSIS OF THIS IS DIFFICULT 3093 02:15:02,840 --> 02:15:07,200 SO WE HAD SUBSTRATES WHERE WE 3094 02:15:07,200 --> 02:15:09,800 HAD A GROUP COULD FLORES AT THE 3095 02:15:09,800 --> 02:15:14,040 RED REGION OF THE SPECTRA AND I 3096 02:15:14,040 --> 02:15:15,480 JUST WANTED TO SHOW YOU WHAT 3097 02:15:15,480 --> 02:15:17,120 HAPPENS WHEN WE DO A SCREENING 3098 02:15:17,120 --> 02:15:18,920 WITH THESE DIFFERENT SOURCES AND 3099 02:15:18,920 --> 02:15:20,720 DIFFERENT SUBSTRATES AND SO HERE 3100 02:15:20,720 --> 02:15:22,360 IS WHAT THE RESULTS OF A 3101 02:15:22,360 --> 02:15:26,040 SCREENING OF A DECK OF 558,000 3102 02:15:26,040 --> 02:15:28,760 COMPOUNDS SO IF WE USE THE WILD 3103 02:15:28,760 --> 02:15:31,480 TYPE PROTEIN WE DISCOVERED A 3104 02:15:31,480 --> 02:15:34,440 NUMBER OF INHIBITORS AND FEW 3105 02:15:34,440 --> 02:15:35,720 ACTIVATORS ON THE OTHER HAND, 3106 02:15:35,720 --> 02:15:38,560 WHEN WE TOOK THE SAME DECK AND 3107 02:15:38,560 --> 02:15:42,960 SCREENED AGAINST THE N370S IN 3108 02:15:42,960 --> 02:15:44,120 THE SPLEEN FROM PATIENTS WE 3109 02:15:44,120 --> 02:15:46,200 DISCOVERED MORE ACTIVATORS AND 3110 02:15:46,200 --> 02:15:49,160 THEN LESS INHIBITORS AND THIS 3111 02:15:49,160 --> 02:15:50,600 SCREENING WAS DONE WITH THIS 3112 02:15:50,600 --> 02:15:52,920 BLUE SUBSTRATE SO IT TELLS YOU, 3113 02:15:52,920 --> 02:15:54,240 DEPENDING ON THE SOURCE OF THE 3114 02:15:54,240 --> 02:15:55,640 ENZYME AND THE SUBSTRATE CHOICE 3115 02:15:55,640 --> 02:15:57,320 YOU HAVE, THE SUBSTRATE CHOICE 3116 02:15:57,320 --> 02:15:58,760 I'LL TALK ABOUT IT IN THE NEXT 3117 02:15:58,760 --> 02:16:00,200 SLIDE, YOU CAN HAVE VERY 3118 02:16:00,200 --> 02:16:03,480 DIFFERENT RESULTS AND THE WAY WE 3119 02:16:03,480 --> 02:16:04,400 RATIONALIZED THIS OUTCOME WAS 3120 02:16:04,400 --> 02:16:06,840 WHEN WE TAKE THE ENZYME WE 3121 02:16:06,840 --> 02:16:10,040 ACTUALLY HAVE TO ADD DETERGENT 3122 02:16:10,040 --> 02:16:13,760 TO MAKE IT ACTIVE AND WE THINK 3123 02:16:13,760 --> 02:16:14,560 THAT THAT ACTIVATES THE ENZYME 3124 02:16:14,560 --> 02:16:17,360 AND PREVENTS YOU FROM 3125 02:16:17,360 --> 02:16:20,680 DISCOVERING ACTIVATORS AND LET 3126 02:16:20,680 --> 02:16:24,360 ME SHOW YOU QUICKLY FOR TYPES WE 3127 02:16:24,360 --> 02:16:26,160 DISCOVERED AND SHOW YOU WHAT 3128 02:16:26,160 --> 02:16:30,760 HAPPENS WHEN WE TAKE JUST THE 3129 02:16:30,760 --> 02:16:31,800 N370S MUTANT, THE COURSE COULD 3130 02:16:31,800 --> 02:16:42,200 BE THE REKUM BAN RE-- AND THE DS 3131 02:16:42,200 --> 02:16:44,440 SHOWN IN LIGHT AND IN BOLD IS 3132 02:16:44,440 --> 02:16:44,800 THE SPLEEN DATA. 3133 02:16:44,800 --> 02:16:47,040 HERE IS THE COMPOUND WHICH SHOWS 3134 02:16:47,040 --> 02:16:49,680 INHIBITION OF THESE TWO 3135 02:16:49,680 --> 02:16:52,840 SUBSTRATES ONLY WHEN YOU USE THE 3136 02:16:52,840 --> 02:16:54,200 SPLEEN. 3137 02:16:54,200 --> 02:16:57,200 THIS COMPOUND SHO SHOWS ACTIVITY 3138 02:16:57,200 --> 02:16:58,520 WHEN YOU USE THE PROTEIN. 3139 02:16:58,520 --> 02:17:00,840 THIS IS GREAT FROM CHEMISTS 3140 02:17:00,840 --> 02:17:03,440 BECAUSE NO MATTER WHAT YOU DO IT 3141 02:17:03,440 --> 02:17:03,920 WORKS. 3142 02:17:03,920 --> 02:17:07,120 IF YOU REMEMBER, THE DISEASE IS 3143 02:17:07,120 --> 02:17:13,200 DUE TO TO DEFICIENCY OF THE PRO. 3144 02:17:13,200 --> 02:17:15,080 WE WANT A COUPLE POUND THAT 3145 02:17:15,080 --> 02:17:17,240 ACTIVATES THE PROTEIN OR AS I 3146 02:17:17,240 --> 02:17:18,680 MENTIONED, INCREASES THE 3147 02:17:18,680 --> 02:17:19,600 CONCENTRATION OF THE PROTEIN AND 3148 02:17:19,600 --> 02:17:21,240 SO THIS WAS A COMPOUND WE 3149 02:17:21,240 --> 02:17:22,440 DISCOVERED WHERE WE SAW 3150 02:17:22,440 --> 02:17:24,320 ACTIVATION WITH RESPECT TO THE 3151 02:17:24,320 --> 02:17:26,000 HYDROLYSIS OF THIS ARTIFICIAL 3152 02:17:26,000 --> 02:17:29,240 SUBSTRATE AND WE SAW ACTIVATION 3153 02:17:29,240 --> 02:17:31,640 ONLY WITH THE BLUE SUBSTRATE. 3154 02:17:31,640 --> 02:17:35,320 AND IN FACT, THIS ASSAY WAS THEN 3155 02:17:35,320 --> 02:17:38,320 UTILIZED TO EXECUTE A MEDICINAL 3156 02:17:38,320 --> 02:17:39,560 CHEM TREE COMPOUND WITH THE 3157 02:17:39,560 --> 02:17:41,360 SPLEEN AND THE BLUE SUBSTRATE 3158 02:17:41,360 --> 02:17:43,080 AND WE DID, WHY WANT TO GO 3159 02:17:43,080 --> 02:17:44,880 THROUGH THIS DATA, WE PUBLISHED 3160 02:17:44,880 --> 02:17:46,320 THIS A LONG TIME BACK BUT WE HAD 3161 02:17:46,320 --> 02:17:51,600 A COMPOUND ML198 WHICH WAS MUCH 3162 02:17:51,600 --> 02:17:53,680 MORE POTENT THAN THE STARTING 3163 02:17:53,680 --> 02:17:54,960 HIT THAT WE HAD AND THEN WE WENT 3164 02:17:54,960 --> 02:17:56,720 ON TO PUBLISH THIS DATA AND THEN 3165 02:17:56,720 --> 02:17:58,240 I'LL SHOW YOU FURTHER 3166 02:17:58,240 --> 02:17:59,600 CHARACTERIZATION WE DID TO THIS 3167 02:17:59,600 --> 02:17:59,840 COMPOUND. 3168 02:17:59,840 --> 02:18:01,840 I WANT TO SHOW YOU THESE 3169 02:18:01,840 --> 02:18:02,800 COMPOUNDS BECAUSE I'M GOING TO 3170 02:18:02,800 --> 02:18:03,840 TALK ABOUT THIS AGAIN. 3171 02:18:03,840 --> 02:18:05,720 WE MADE THESE COMPOUNDS WHERE WE 3172 02:18:05,720 --> 02:18:08,160 REPLACED THE RING WITH FOUND 3173 02:18:08,160 --> 02:18:10,440 THEY HAD NOT THAT GREAT POTENCY 3174 02:18:10,440 --> 02:18:12,560 BUT THEY WERE IMPORTANT LATER 3175 02:18:12,560 --> 02:18:12,760 ON. 3176 02:18:12,760 --> 02:18:16,200 SO WE TOOK THIS COMPOUND AND 3177 02:18:16,200 --> 02:18:17,480 ACTUALLY SHOWED THAT NOT ONLY IS 3178 02:18:17,480 --> 02:18:19,280 THE COMPOUND ABLE TO ACTUALLY 3179 02:18:19,280 --> 02:18:23,480 INCREASE THE CONCENTRATION OF 3180 02:18:23,480 --> 02:18:24,920 PATIENT DERIVED MACROPHAGES. 3181 02:18:24,920 --> 02:18:26,800 SO MACROPHAGES ARE THE CELLS 3182 02:18:26,800 --> 02:18:28,360 WHERE THE DISEASE ACTUALLY 3183 02:18:28,360 --> 02:18:32,160 HAPPENS AND WHERE WE SEE VERY 3184 02:18:32,160 --> 02:18:35,160 LOW QUANTITIES OF IN GROWN AND 3185 02:18:35,160 --> 02:18:37,160 WHEN YOU ADD THE COMPOUND YOU 3186 02:18:37,160 --> 02:18:38,360 CAN SEE THE GREEN GOES TO THE 3187 02:18:38,360 --> 02:18:39,880 RED THAT'S WHY YOU HAVE THIS 3188 02:18:39,880 --> 02:18:41,800 YELLOW SIGNAL SO WITH THE 3189 02:18:41,800 --> 02:18:45,080 COMPOUND INCUBATION WE SEE A 3190 02:18:45,080 --> 02:18:46,640 PROFOUND INCREASE IN THE 3191 02:18:46,640 --> 02:18:52,440 CONCENTRATION OF THE ENZYME IN 3192 02:18:52,440 --> 02:18:53,520 THE LICE OWE SOMEWHERE WE WANT 3193 02:18:53,520 --> 02:18:56,200 IT TO GO. 3194 02:18:56,200 --> 02:19:04,960 THERE'S AN OWESSIZATION OF ASSON 3195 02:19:04,960 --> 02:19:07,040 PARKINSON'S DISEASE, WE TOOK 3196 02:19:07,040 --> 02:19:08,120 SHELLS FROM PARKINSON PATIENTS 3197 02:19:08,120 --> 02:19:11,360 AND FIBER BLAST AND GENERATE 3198 02:19:11,360 --> 02:19:14,840 IPSCs AND DIFFERENTIATE THEM 3199 02:19:14,840 --> 02:19:16,440 INTO NEURONS AND WE WERE ABLE TO 3200 02:19:16,440 --> 02:19:19,040 SHOW THIS COMPOUND IS ABLE TO 3201 02:19:19,040 --> 02:19:21,800 CLEAR AGO AGREE GATES FROM THESE 3202 02:19:21,800 --> 02:19:23,480 NEURONS AND I CAN TALK ABOUT 3203 02:19:23,480 --> 02:19:26,120 THIS IF YOU GUYS ARE INTERESTED. 3204 02:19:26,120 --> 02:19:28,080 ESSENTIALLY WHAT I'M SHOWING 3205 02:19:28,080 --> 02:19:28,920 YOU. 3206 02:19:28,920 --> 02:19:30,160 THESE ASSAYS ARE DIFFICULT TO DO 3207 02:19:30,160 --> 02:19:31,360 AND YOU HAVE TO AGE THE NEURONS 3208 02:19:31,360 --> 02:19:32,800 FOR SEVERAL DAYS TO SEE THE 3209 02:19:32,800 --> 02:19:36,000 PHONE OWE TYPE AND WHAT OUR 3210 02:19:36,000 --> 02:19:37,400 BIOLOGY COLLEAGUES WERE ABLE TO 3211 02:19:37,400 --> 02:19:41,880 SHOW THIS COMPOUND ACTUALLY IS 3212 02:19:41,880 --> 02:19:44,360 ABLE TO DO THIS PROFOUND 3213 02:19:44,360 --> 02:19:50,040 REDUCTION OF AGO AGREE GATES. 3214 02:19:50,040 --> 02:19:52,040 IT WAS LICENSED TO ACCOMPANY AND 3215 02:19:52,040 --> 02:19:54,040 THEY MODIFIED THIS TO THIS 3216 02:19:54,040 --> 02:19:55,200 CLINICAL COMPOUND WHICH WENT TO 3217 02:19:55,200 --> 02:20:05,720 THE CLINIC AND I'M LOOKING TO 3218 02:20:08,760 --> 02:20:11,360 SEE WHAT HAPPENS WITH THIS CHEMO 3219 02:20:11,360 --> 02:20:11,960 TYPE. 3220 02:20:11,960 --> 02:20:14,200 SORRY, I HAVE TO ADVERTISE THIS. 3221 02:20:14,200 --> 02:20:18,400 SO THIS IS AN ARTICLE I ACTUALLY 3222 02:20:18,400 --> 02:20:20,880 WROTE WITH POSTDOCTORAL TRAINEES 3223 02:20:20,880 --> 02:20:27,280 IN MY GROUP. 3224 02:20:27,280 --> 02:20:29,440 WE WENT THROUGH SIMILAR CASE 3225 02:20:29,440 --> 02:20:31,280 STUDIES TO SHOWCASE THAT THE 3226 02:20:31,280 --> 02:20:33,680 ASSAY IS IMPORTANT IN DEFINING 3227 02:20:33,680 --> 02:20:35,400 THE BIOLOGY OF YOUR COMPOUNDS 3228 02:20:35,400 --> 02:20:37,400 AND LASTLY, I JUST WANT TO HIT 3229 02:20:37,400 --> 02:20:40,040 HOME THE SAME MESSAGE THAT YOU 3230 02:20:40,040 --> 02:20:41,560 KNOW, ASSAYS CAN BE REALLY 3231 02:20:41,560 --> 02:20:44,160 IMPORTANT AND WORK WITH YOUR 3232 02:20:44,160 --> 02:20:45,080 BIOLOGIST TO UNDERSTAND THEM. 3233 02:20:45,080 --> 02:20:46,520 THEY CAN MAKE OR BREAK A 3234 02:20:46,520 --> 02:20:47,720 CAMPAIGN AND OBVIOUSLY, I HAVE 3235 02:20:47,720 --> 02:20:49,040 TO THANK A NUMBER OF PEOPLE WHO 3236 02:20:49,040 --> 02:20:50,400 ARE WORKING THESE PROJECTS AND 3237 02:20:50,400 --> 02:20:55,200 THANK YOU FOR YOUR TIME. 3238 02:20:55,200 --> 02:20:58,440 >>THANK YOU SO MUCH, SAM, FOR A 3239 02:20:58,440 --> 02:20:59,880 WONDERFUL TALK AS USUAL. 3240 02:20:59,880 --> 02:21:04,040 WE HAVE QUESTIONS HERE. 3241 02:21:04,040 --> 02:21:09,680 >>THANK YOU FOR A WONDERFUL AND 3242 02:21:09,680 --> 02:21:19,000 JUST REFERENCES WOULD THAT 3243 02:21:19,000 --> 02:21:29,520 FOLLOW THE SAME PRINCIPLES THE 3244 02:21:35,280 --> 02:21:36,200 CONTRACTOR, THE CONCEPT WHERE 3245 02:21:36,200 --> 02:21:39,280 YOU LOOK AT THINGS LIKE, YOU 3246 02:21:39,280 --> 02:21:41,120 KNOW, WHERE YOU HAVE A KINASE 3247 02:21:41,120 --> 02:21:46,560 THAT PHOSPHORYLATION A BUNCH OF 3248 02:21:46,560 --> 02:21:48,160 DIFFERENT KINASES TO SEE HOW 3249 02:21:48,160 --> 02:21:50,760 THEY'RE IMPACTED WITH THE DRUG. 3250 02:21:50,760 --> 02:21:55,080 SIMILARLY, WOULD THAT APPLY TO 3251 02:21:55,080 --> 02:21:57,800 THE POST TRANSLATIONAL 3252 02:21:57,800 --> 02:22:00,440 MODIFICATION AS WELL OR IS IT 3253 02:22:00,440 --> 02:22:01,680 JUST FOR PHOSPHORYLATION. 3254 02:22:01,680 --> 02:22:03,960 >>IF THERE'S BIOLOGY THAT LOOKS 3255 02:22:03,960 --> 02:22:05,240 AT DOWNSTREAM EVENTS, I WOULD 3256 02:22:05,240 --> 02:22:07,640 ALWAYS PREFER THAT WE MONITOR 3257 02:22:07,640 --> 02:22:12,000 THAT SO WHEN YOU ARE ESSENTIALLY 3258 02:22:12,000 --> 02:22:13,080 OPTIMIZING COMPOUNDS WHICH ARE 3259 02:22:13,080 --> 02:22:18,760 DIRECTLY BINDING WITH THE 3260 02:22:18,760 --> 02:22:20,120 TARGET, THE DOWNSTREAM EVENTS 3261 02:22:20,120 --> 02:22:21,240 TAKE PLACE AS WELL. 3262 02:22:21,240 --> 02:22:22,160 SOMETIMES IT CAN BE DIFFICULT 3263 02:22:22,160 --> 02:22:29,680 BECAUSE DESIGNING ASSAYS FOR 3264 02:22:29,680 --> 02:22:32,840 DOWNSTREAM AT LEAST TARGET 3265 02:22:32,840 --> 02:22:35,080 ENGAGEMENT IF THE COMPOUND IS 3266 02:22:35,080 --> 02:22:36,520 MAKING IS ENGAGING WITH THE 3267 02:22:36,520 --> 02:22:38,400 TARGET BECAUSE YOU CAN DO ASSAYS 3268 02:22:38,400 --> 02:22:42,840 IN HIGH-THROUGHPUT WE HAVE FOLKS 3269 02:22:42,840 --> 02:22:53,000 HERE WHO IT COULD BE LIMITEDDED 3270 02:22:53,000 --> 02:23:01,120 WHAT IS AVAILABLE TO YOU AND 3271 02:23:01,120 --> 02:23:11,240 YEAH. 3272 02:23:12,440 --> 02:23:13,800 >>DO YOU LIKE AT WHAT WAS 3273 02:23:13,800 --> 02:23:14,640 DRIVING THAT? 3274 02:23:14,640 --> 02:23:16,600 >>THAT'S A GREAT QUESTION, THIS 3275 02:23:16,600 --> 02:23:18,080 IS SOMETHING THAT PEOPLE OFTEN 3276 02:23:18,080 --> 02:23:20,520 ASKED ME BECAUSE I SHOWED 3277 02:23:20,520 --> 02:23:22,080 DIFFERENT ENZYME SOURCES AND 3278 02:23:22,080 --> 02:23:25,840 JUST WHAT IS THE STRUCTURAL 3279 02:23:25,840 --> 02:23:28,920 BASIS AND IN THE KINASE ASSAY, 3280 02:23:28,920 --> 02:23:31,600 WE, I CAN SPECULATE THAT PERHAPS 3281 02:23:31,600 --> 02:23:33,400 IN THIS THE CELLULAR ENVIRONMENT 3282 02:23:33,400 --> 02:23:35,800 IT'S MORE PHYSIOLOGICALLY 3283 02:23:35,800 --> 02:23:39,520 RELEVANT AND IT'S PROBABLY 3284 02:23:39,520 --> 02:23:49,720 INTERACTING. 3285 02:23:50,480 --> 02:23:54,800 YEAH -- I WOULD BELIEVE THE CELL 3286 02:23:54,800 --> 02:23:55,720 BASE YEAH. 3287 02:23:55,720 --> 02:23:57,320 THEY HAVE MARK HERE WHO WAS 3288 02:23:57,320 --> 02:23:59,560 HEADING THE BIOLOGY THERE AND HE 3289 02:23:59,560 --> 02:24:01,360 CAN TELL YOU WHAT HE BELIEVES. 3290 02:24:01,360 --> 02:24:04,920 YEAH, I WOULD BELIEVE THAT. 3291 02:24:04,920 --> 02:24:10,520 [OFF MIC] 3292 02:24:10,520 --> 02:24:12,160 >>YES, THAT IS TRUE. 3293 02:24:12,160 --> 02:24:15,360 YOU ARE RIGHT. 3294 02:24:15,360 --> 02:24:16,440 I DON'T THINK -- THAT'S WHY I 3295 02:24:16,440 --> 02:24:18,560 HAVE TO WORK WITH PEOPLE LIKE 3296 02:24:18,560 --> 02:24:18,880 YOU. 3297 02:24:18,880 --> 02:24:21,520 THE QUESTION WAS HERE, NATHAN 3298 02:24:21,520 --> 02:24:24,080 SAID, THAT THE ATP CONCENTRATION 3299 02:24:24,080 --> 02:24:26,520 AND THE BIOCHEMICAL ASSAY COULD 3300 02:24:26,520 --> 02:24:28,360 BE IMPORTANT AND MAYBE THAT WAS 3301 02:24:28,360 --> 02:24:30,120 ONE OF THE REASONS WHY WE DIDN'T 3302 02:24:30,120 --> 02:24:31,440 SEE ACTIVITY AGAINST THE TWO 3303 02:24:31,440 --> 02:24:35,400 OTHER BIOLOGICAL TARGETS. 3304 02:24:35,400 --> 02:24:36,720 THAT IS WHY YOU HAVE TO WORK 3305 02:24:36,720 --> 02:24:40,320 WITH FOLKS LIKE YOU. 3306 02:24:40,320 --> 02:24:50,800 WHO WILL READ THE PORT CAL. 3307 02:25:00,320 --> 02:25:03,000 [OFF MIC] 3308 02:25:03,000 --> 02:25:36,520 YES. 3309 02:25:36,520 --> 02:25:39,880 [OFF MIC] 3310 02:25:39,880 --> 02:25:40,040 YES. 3311 02:25:40,040 --> 02:25:44,240 SO THIS IS A VERY IMPORTANT 3312 02:25:44,240 --> 02:25:48,040 POINT. 3313 02:25:48,040 --> 02:25:50,240 SOMETIMES THE FOLKS WHO ACTUALLY 3314 02:25:50,240 --> 02:25:51,760 DEVELOP THESE PLATFORMS, THEY 3315 02:25:51,760 --> 02:25:54,040 WILL LOOK AT SPECIFIC SEQUENCES 3316 02:25:54,040 --> 02:25:57,040 OF PEPTIDES OF THE SUBSTRATE AND 3317 02:25:57,040 --> 02:25:58,120 DESIGN THE PEPTIDE SEQUENCE. 3318 02:25:58,120 --> 02:26:00,920 IF YOU HAVE THAT IT'S MUCH MORE 3319 02:26:00,920 --> 02:26:01,880 RELEVANT THAN SUBSTRATE. 3320 02:26:01,880 --> 02:26:05,320 SOMETIMES YOU GO WITH WHAT YOU 3321 02:26:05,320 --> 02:26:05,520 HAVE. 3322 02:26:05,520 --> 02:26:11,800 [OFF MIC] 3323 02:26:11,800 --> 02:26:14,520 >>YES. 3324 02:26:14,520 --> 02:26:17,160 SO THAT'S A VERY -- YOU PICKED 3325 02:26:17,160 --> 02:26:18,560 THAT UP. 3326 02:26:18,560 --> 02:26:22,200 SO, EVEN WITHOUT THE DIMER, YOU 3327 02:26:22,200 --> 02:26:27,440 CAN ADD IN A SUBSTRATE AND IT 3328 02:26:27,440 --> 02:26:29,440 FOSS FOR LATES AND IT DOES THE 3329 02:26:29,440 --> 02:26:33,880 AUTO PHOSPHORYLATION. 3330 02:26:33,880 --> 02:26:34,160 WONDERFUL. 3331 02:26:34,160 --> 02:26:35,680 THANK YOU SO MUCH AGAIN, SAM. 3332 02:26:35,680 --> 02:26:43,080 [APPLAUSE] 3333 02:26:43,080 --> 02:26:44,040 ALL RIGHT. 3334 02:26:44,040 --> 02:26:52,840 OUR NEXT SPEAKER IS Dr. SOFIE 3335 02:26:52,840 --> 02:26:54,560 A SENIOR SCIENTIST AT PFIZER 3336 02:26:54,560 --> 02:26:58,440 WITH WHERE SHE SUPPOSE SCREENING 3337 02:26:58,440 --> 02:26:59,640 PLATFORM AND HIT DISCOVERY 3338 02:26:59,640 --> 02:27:02,800 WITHIN THE PRIMARY PHARMACOLOGY 3339 02:27:02,800 --> 02:27:03,160 GROUP. 3340 02:27:03,160 --> 02:27:05,160 HER TALK TODAY IS ENTITLED 3341 02:27:05,160 --> 02:27:09,320 BUILDING THE EFFICIENT DISCOVERY 3342 02:27:09,320 --> 02:27:10,960 OF ACTIONABLE CHEMICAL MATTER 3343 02:27:10,960 --> 02:27:14,080 FROM DNA-EN CODED LIBRARIES. 3344 02:27:14,080 --> 02:27:24,520 THANK YOU SO MUCH, SOPHIA. 3345 02:27:30,040 --> 02:27:31,360 >>THANK YOU FOR THE KIND 3346 02:27:31,360 --> 02:27:32,240 INTRODUCTION AND THANK YOU FOR 3347 02:27:32,240 --> 02:27:34,480 GIVING ME THE OPPORTUNITY TO BE 3348 02:27:34,480 --> 02:27:35,800 HERE AT THIS WORKSHOP. 3349 02:27:35,800 --> 02:27:38,720 AS THE TITLE INDICATES, MY TALK 3350 02:27:38,720 --> 02:27:40,200 BEFORE WAS ON THE UNCODED 3351 02:27:40,200 --> 02:27:41,800 LIBRARY SCREENING AND THE WORK 3352 02:27:41,800 --> 02:27:44,720 FLOW THAT THE PFIZER GROUP 3353 02:27:44,720 --> 02:27:45,720 IMPLEMENTED IN THE LAST COUPLE 3354 02:27:45,720 --> 02:27:47,120 OF YEARS TO ENSURE THE 3355 02:27:47,120 --> 02:27:48,800 ACTIONABLE CHEMICAL MATTER AT 3356 02:27:48,800 --> 02:27:50,640 THE END OF THE SCREENING. 3357 02:27:50,640 --> 02:27:53,480 NOW BEFORE I START, HOW MANY OF 3358 02:27:53,480 --> 02:27:54,760 YOU ARE FAMILIAR WITH THE 3359 02:27:54,760 --> 02:27:57,600 SCREENING, RAISE YOUR HANDS? 3360 02:27:57,600 --> 02:27:59,800 A FEW OF YOU, THAT'S NOT BAD. 3361 02:27:59,800 --> 02:28:00,800 SO FOR PEOPLE WHO ARE NOT 3362 02:28:00,800 --> 02:28:02,040 FAMILIAR WITH IT DON'T WORRY 3363 02:28:02,040 --> 02:28:03,400 BECAUSE I'M GOING TO START MY 3364 02:28:03,400 --> 02:28:05,320 TALK WITH A BRIEF INTRODUCTION 3365 02:28:05,320 --> 02:28:06,920 TO THE SCREENING. 3366 02:28:06,920 --> 02:28:08,440 AND AFTER THAT, I'LL SHOW YOU 3367 02:28:08,440 --> 02:28:11,840 HOW WE AT PFIZER VALIDATE OUR 3368 02:28:11,840 --> 02:28:14,760 PROTEIN REAGENTS BEFORE THE 3369 02:28:14,760 --> 02:28:16,320 SELECTION AND LASTLY I'LL TALK 3370 02:28:16,320 --> 02:28:18,200 ABOUT THE WORK FLOW THAT WE 3371 02:28:18,200 --> 02:28:20,280 IMPLEMENTED TO ENSURE THAT WE 3372 02:28:20,280 --> 02:28:22,200 HAVE HIGH HEAT CONFIRMATION 3373 02:28:22,200 --> 02:28:24,200 RATES WHEN WE SYNTHESIZE THE 3374 02:28:24,200 --> 02:28:27,800 COMPOUNDS OF DNA AND ENSURE THE 3375 02:28:27,800 --> 02:28:31,000 COMPOUNDS HAVE GOOD DRUG-LIKE 3376 02:28:31,000 --> 02:28:32,600 PROPERTIES AND HOPEFULLY IN THE 3377 02:28:32,600 --> 02:28:34,640 LAST COUPLE OF MINUTES I'LL TALK 3378 02:28:34,640 --> 02:28:38,400 ABOUT CELL BASED SCREENING AS 3379 02:28:38,400 --> 02:28:41,800 WELL AND MEMBRANE PROTEINS. 3380 02:28:41,800 --> 02:28:43,680 LET ME START WITH A BRIEF 3381 02:28:43,680 --> 02:28:44,160 INTRODUCTION. 3382 02:28:44,160 --> 02:28:47,280 SO, THE DNA-ENCODED LIBRARY 3383 02:28:47,280 --> 02:28:49,160 SCREENING IS A RELATIVELY NEW 3384 02:28:49,160 --> 02:28:49,480 TECHNOLOGY. 3385 02:28:49,480 --> 02:28:51,520 IT WAS DEVELOPED IN THE LAST 3386 02:28:51,520 --> 02:28:52,920 20-25 YEARS BUT IT REALLY CAUGHT 3387 02:28:52,920 --> 02:28:55,680 ON IN THE LAST DECADE OR SO. 3388 02:28:55,680 --> 02:28:57,960 SO, THOSE ARE A COLLECTION OF 3389 02:28:57,960 --> 02:29:01,080 COMPOUNDS THAT ARE LINKED TO A 3390 02:29:01,080 --> 02:29:03,840 DNA TAG AND SO, EVERY MOLECULE 3391 02:29:03,840 --> 02:29:05,960 IN YOUR LIBRARY WHICH WE CALL 3392 02:29:05,960 --> 02:29:08,520 WAR HEADS HAS A UNIQUE DNA TAG 3393 02:29:08,520 --> 02:29:12,320 WHO SEQUENCES DESCRI DESCRIBES E 3394 02:29:12,320 --> 02:29:14,000 BUILDING BLOCKS IN THE LIBRARY 3395 02:29:14,000 --> 02:29:16,080 SO FOR EXAMPLE THE BLUE BUILDING 3396 02:29:16,080 --> 02:29:18,440 BLOCKS IS ENCODED BY THE BLUE 3397 02:29:18,440 --> 02:29:20,000 SEQUENCE AND THE RED BY THE RED 3398 02:29:20,000 --> 02:29:22,760 SEQUENCE AND SO ON. 3399 02:29:22,760 --> 02:29:26,800 NOW, MY TALK WILL FOCUS ON AND 3400 02:29:26,800 --> 02:29:28,920 SCREENING AND NOT SYNTHESIS BUT 3401 02:29:28,920 --> 02:29:30,560 STILL I WANTED TO SHOW YOU HOW 3402 02:29:30,560 --> 02:29:31,960 THESE COMPOUNDS ARE MADE BECAUSE 3403 02:29:31,960 --> 02:29:41,720 I THINK IT'S QUITE WE REFER TO 3404 02:29:41,720 --> 02:29:43,760 THIS A METHODOLOGY SO WE START 3405 02:29:43,760 --> 02:29:47,320 WITH A DNA HAT PIECE KICKED TO A 3406 02:29:47,320 --> 02:29:48,440 LINKER AND WE SPLIT THAT AND 3407 02:29:48,440 --> 02:29:52,280 PLACE IT INTO A MORE PLATE AND 3408 02:29:52,280 --> 02:29:53,720 LET'S SAY A PLATE. 3409 02:29:53,720 --> 02:29:56,320 NOW, AFTER THAT, WE DO OUR FIRST 3410 02:29:56,320 --> 02:29:58,360 LIGATION OF THE DNA TAG. 3411 02:29:58,360 --> 02:30:01,360 SO, AT THIS STEP, EVERY SINGLE 3412 02:30:01,360 --> 02:30:03,560 WELL WE RECEIVED A UNIQUE DNA 3413 02:30:03,560 --> 02:30:05,520 SEQUENCE SO AFTER YOUR LIGATION, 3414 02:30:05,520 --> 02:30:09,720 YOU END UP WITH 96 UNIQUE DNA 3415 02:30:09,720 --> 02:30:10,200 SEQUENCES. 3416 02:30:10,200 --> 02:30:12,000 SO WHEN YOU LIGATE YOUR FIRST 3417 02:30:12,000 --> 02:30:13,800 DNA TAG, YOU ADD YOUR FIRST 3418 02:30:13,800 --> 02:30:15,840 BUILDING BLOCK AND AGAIN, EACH 3419 02:30:15,840 --> 02:30:17,480 WELL WE RECEIVED THE FIRST 3420 02:30:17,480 --> 02:30:20,240 BUILDING BLOCKS SO AFTER YOUR 3421 02:30:20,240 --> 02:30:22,080 SYNTHESIS, YOU WIND UP WITH 96 3422 02:30:22,080 --> 02:30:24,160 DIFFERENT COMPOUNDS WITH ITS OWN 3423 02:30:24,160 --> 02:30:26,040 UNIQUE DNA SEQUENCES. 3424 02:30:26,040 --> 02:30:30,480 SO AFTER THIS SYNTHESIS, YOU 3425 02:30:30,480 --> 02:30:32,520 POOL ALL YOUR TOGETHER INTO A 3426 02:30:32,520 --> 02:30:38,200 TUBE AND MIX THEM EFFICIENTLY 3427 02:30:38,200 --> 02:30:39,320 AND REDISTRIBUTE THEM. 3428 02:30:39,320 --> 02:30:41,120 EVERY SINGLE WELL IN YOUR PLATE 3429 02:30:41,120 --> 02:30:42,480 WILL HAVE ALL THE COMPOUNDS THAT 3430 02:30:42,480 --> 02:30:44,720 YOU JUST SYNTHESIZED IN THE 3431 02:30:44,720 --> 02:30:45,400 FIRST ROUND. 3432 02:30:45,400 --> 02:30:47,200 AND THEN YOU REPEAT THIS CYCLE A 3433 02:30:47,200 --> 02:30:48,040 COUPLE MORE TIMES. 3434 02:30:48,040 --> 02:30:49,960 SO AGAIN, YOU ARE GOING TO ADD 3435 02:30:49,960 --> 02:30:52,440 YOUR SECOND DNA TAG, YOUR 3436 02:30:52,440 --> 02:30:53,720 BUILDING BUILDING BLOCK AND 3437 02:30:53,720 --> 02:30:56,080 EVERY SINGLE WILL RECEIVE ITS 3438 02:30:56,080 --> 02:30:58,000 UNIQUE BLOCKS AND SO YOU REPEAT 3439 02:30:58,000 --> 02:30:59,160 THIS TWO OR THREE TIMES 3440 02:30:59,160 --> 02:31:00,600 DEPENDING ON THE COMPOUNDS THAT 3441 02:31:00,600 --> 02:31:02,400 YOU WANT TO SYNTHESIZE SO FOR 3442 02:31:02,400 --> 02:31:03,840 EXAMPLE, AT THE END HERE, YOU 3443 02:31:03,840 --> 02:31:06,000 WILL HAVE A COMPOUND WITH THROW 3444 02:31:06,000 --> 02:31:06,920 DIFFERENT BUILDING BLOCKS AND 3445 02:31:06,920 --> 02:31:09,560 IT'S OWN UNIQUE DNA TAG. 3446 02:31:09,560 --> 02:31:12,160 NOW JUST SOMETHING TO KEEP IN 3447 02:31:12,160 --> 02:31:14,400 MIND, SINCE WE'RE DOING ORGAN 3448 02:31:14,400 --> 02:31:16,480 SYNTHESIS IN THE PRESENCE OF DNA 3449 02:31:16,480 --> 02:31:18,200 WE HAVE TO HAVE WATER PRESENT SO 3450 02:31:18,200 --> 02:31:20,680 IT'S POSSIBLE THAT UNDER THESE 3451 02:31:20,680 --> 02:31:22,240 CONDITIONS, NOT EVERY CHEMICAL 3452 02:31:22,240 --> 02:31:23,800 REACTION WE PROCEED AS EXPECTED, 3453 02:31:23,800 --> 02:31:25,040 AND SOME OF YOUR BUILDING BLOCKS 3454 02:31:25,040 --> 02:31:27,920 JUST WON'T BE INCORPORATED. 3455 02:31:27,920 --> 02:31:29,560 SO, THE NICE THING ABOUT THE 3456 02:31:29,560 --> 02:31:30,960 SPLIT AND PULL TECHNOLOGY THAT 3457 02:31:30,960 --> 02:31:35,600 JUST BY USING A SINGLE PLATE WE 3458 02:31:35,600 --> 02:31:37,120 COULD MAKE A MILLION COMPOUNDS 3459 02:31:37,120 --> 02:31:39,600 AND WE REALLY NEEDED JUST 96 FOR 3460 02:31:39,600 --> 02:31:43,000 THAT AND NOT ONE MILLION 3461 02:31:43,000 --> 02:31:43,360 DIFFERENT VIALS. 3462 02:31:43,360 --> 02:31:47,760 NOW, AGAIN, I'M NOT GOING TO 3463 02:31:47,760 --> 02:31:49,000 TALK ABOUT DELL SYNTHESIS AND 3464 02:31:49,000 --> 02:31:51,280 GREAT PUBLICATIONS OUT THERE 3465 02:31:51,280 --> 02:31:56,800 THAT YOU COULD JUST READ UP ON. 3466 02:31:56,800 --> 02:32:00,640 THE LIBRARIES ARE POISE TOGETHER 3467 02:32:00,640 --> 02:32:05,160 AND THIS IS WHAT WE USE FOR 3468 02:32:05,160 --> 02:32:15,720 SCREENING AND SO WE INCUBATE THE 3469 02:32:23,080 --> 02:32:24,960 PROTEIN AND WHEN YOU REACH 3470 02:32:24,960 --> 02:32:26,440 BINDING EQUITY BIOME YOU CAN 3471 02:32:26,440 --> 02:32:28,480 JUST USE THE MAGNET AND PULL 3472 02:32:28,480 --> 02:32:30,760 EVERYTHING DOWN, YOU PULL DOWN 3473 02:32:30,760 --> 02:32:33,080 YOUR BEADS, PROTEIN, BIO 3474 02:32:33,080 --> 02:32:36,440 COMPOUNDS AND REMOVE THE UNBOUND 3475 02:32:36,440 --> 02:32:36,720 COMPOUNDS. 3476 02:32:36,720 --> 02:32:39,480 AFTER THAT, YOU TAKE YOUR BEADS 3477 02:32:39,480 --> 02:32:40,600 AND YOUR PROTEINS THROUGH A 3478 02:32:40,600 --> 02:32:45,120 COUPLE OF WASH CYCLES TO REMOVE 3479 02:32:45,120 --> 02:32:55,960 THE BINDE BINDERS SO WE RUN A CE 3480 02:32:59,720 --> 02:33:02,240 OF SELECTIONS WHICH MEANS THAT 3481 02:33:02,240 --> 02:33:03,560 WE TAKE THESE SPECIFIC ALLUDED 3482 02:33:03,560 --> 02:33:05,320 COMPOUNDS FROM ROUND ONE AND WE 3483 02:33:05,320 --> 02:33:08,000 TAKE THEM INTO ROUND TWO AS 3484 02:33:08,000 --> 02:33:09,760 INPUT COMPOUNDS. 3485 02:33:09,760 --> 02:33:11,280 AND RUNNING A COUPLE ROUNDS OF 3486 02:33:11,280 --> 02:33:12,960 SELECTION IS NECESSARY BECAUSE 3487 02:33:12,960 --> 02:33:17,320 YOU WANT TO REACH YOUR SPECIFIC 3488 02:33:17,320 --> 02:33:18,840 LIG ANTS ENOUGH AND REUSE YOUR 3489 02:33:18,840 --> 02:33:22,040 BACKGROUND SO YOU CAN DOSE YOUR 3490 02:33:22,040 --> 02:33:24,880 COMPOUNDS ON A SEQUENCING CHIP. 3491 02:33:24,880 --> 02:33:28,160 NOW, THE SELECTION TAKES A 3492 02:33:28,160 --> 02:33:29,720 COUPLE OF HOURS BUT WE DON'T 3493 02:33:29,720 --> 02:33:31,840 HAVE A SCIENTIST CHAINED TO THE 3494 02:33:31,840 --> 02:33:33,000 BENCH AND RUNNING THIS FOR HOURS 3495 02:33:33,000 --> 02:33:33,600 AND HOURS. 3496 02:33:33,600 --> 02:33:35,400 AT THIS POINT, EVERYTHING IS 3497 02:33:35,400 --> 02:33:36,840 AUTOMATED SO YOU LITERALLY JUST 3498 02:33:36,840 --> 02:33:39,200 TAKE YOUR PLATE, PIPE IT IN YOUR 3499 02:33:39,200 --> 02:33:40,640 REACH AND THE REST IS TAKEN CARE 3500 02:33:40,640 --> 02:33:43,480 OF BY AN INSTRUMENT. 3501 02:33:43,480 --> 02:33:45,000 THE ADVANTAGE OF -- THEY HAVE 3502 02:33:45,000 --> 02:33:46,880 SEVERAL ADVANTAGES COMPARED TO 3503 02:33:46,880 --> 02:33:49,240 TRADITIONAL HDS FOR EXAMPLE WE 3504 02:33:49,240 --> 02:33:51,120 REQUIRE LESS PROTEIN REAGENT 3505 02:33:51,120 --> 02:33:53,400 COMPARED TO HDS IN OUR CASE, WE 3506 02:33:53,400 --> 02:33:55,240 USUALLY ASK FOR TWO OR THROW 3507 02:33:55,240 --> 02:33:56,560 MILL A GRAMS OF PROTEIN AND THAT 3508 02:33:56,560 --> 02:33:58,320 IS ENOUGH FOR REGION VALIDATION 3509 02:33:58,320 --> 02:34:01,760 AND FOR SCREENING AND FOR HIT 3510 02:34:01,760 --> 02:34:03,880 VALIDATION AND FOR THE 3511 02:34:03,880 --> 02:34:08,840 OCCASIONAL MESS UPS AND ASSAY 3512 02:34:08,840 --> 02:34:10,640 REPEATS. 3513 02:34:10,640 --> 02:34:11,440 NOW, ALSO DELL LIBRARIES DON'T 3514 02:34:11,440 --> 02:34:13,680 HAVE ANY BIAS AGAINST THEM. 3515 02:34:13,680 --> 02:34:16,920 AND WHAT I MEAN BY THAT, IS FOR 3516 02:34:16,920 --> 02:34:18,040 EXAMPLE, IF YOUR COMPANY HAS 3517 02:34:18,040 --> 02:34:20,160 BEEN HEAVILY WORKED IN 3518 02:34:20,160 --> 02:34:22,080 NEUROSCIENCE SPACE, YOU PROBABLY 3519 02:34:22,080 --> 02:34:25,720 HAVE A CHEMICAL MATTER WHICH HAS 3520 02:34:25,720 --> 02:34:28,560 BIAS AGAINST COMPOUNDS THAT HAVE 3521 02:34:28,560 --> 02:34:29,520 SPECIFIC PHYSICAL CHEMICAL 3522 02:34:29,520 --> 02:34:31,480 PROPERTIES THAT ALLOWS THEM TO 3523 02:34:31,480 --> 02:34:33,480 CROSS THE BROAD BAIN BARRIERS 3524 02:34:33,480 --> 02:34:35,640 AND YOUR CHEMICAL METHOD DOESN'T 3525 02:34:35,640 --> 02:34:36,960 COVER ALL THE OTHER CHEMICAL 3526 02:34:36,960 --> 02:34:39,080 SPACE HOWEVER THIS IS NOT TRUE 3527 02:34:39,080 --> 02:34:41,000 FOR DEL SO THEY DON'T HAVE BIAS 3528 02:34:41,000 --> 02:34:42,560 AGAINST THEM AND THEY COVER A 3529 02:34:42,560 --> 02:34:45,360 WIDE CHEMICAL SPACE. 3530 02:34:45,360 --> 02:34:47,720 NOW, DEL VALIDATION AND 3531 02:34:47,720 --> 02:34:49,760 SCREENING ITSELF IS RELATIVELY 3532 02:34:49,760 --> 02:34:51,320 FAST AND WE CAN RUN THESE IN 3533 02:34:51,320 --> 02:34:53,440 JUST A COUPLE OF WEEKS. 3534 02:34:53,440 --> 02:34:58,160 AND ALSO, WE CAN PENALIZE 3535 02:34:58,160 --> 02:34:59,720 MULTIPLE CONDITIONS BUT I'LL GET 3536 02:34:59,720 --> 02:35:02,080 BACK TO THAT A LITTLE BIT LATER. 3537 02:35:02,080 --> 02:35:05,160 BEFORE I GET INTO THE DETAILS, 3538 02:35:05,160 --> 02:35:07,240 OF SCREENING AND THESE ANALYZES, 3539 02:35:07,240 --> 02:35:09,280 LET ME JUST WALK YOU THROUGH 3540 02:35:09,280 --> 02:35:12,960 WHAT IT TAKES TO BE A HIT IN DEL 3541 02:35:12,960 --> 02:35:14,520 SELECTION AND WHAT IS EXPECTED 3542 02:35:14,520 --> 02:35:16,080 OUTCOME THAT YOU WOULD SEE BASED 3543 02:35:16,080 --> 02:35:18,320 ON THE BIO CHEM TREES THAT IS 3544 02:35:18,320 --> 02:35:25,720 GOING ON IN YOUR BINDING 3545 02:35:25,720 --> 02:35:35,720 REACTION. 3546 02:35:35,720 --> 02:35:37,960 IN BOTH CASES, THE CONCENTRATION 3547 02:35:37,960 --> 02:35:40,000 OF EITHER THE LIG ANT OR THE 3548 02:35:40,000 --> 02:35:42,120 PROTEIN HAS TO EXCEED THE KD OF 3549 02:35:42,120 --> 02:35:43,520 THE INTERACTION. 3550 02:35:43,520 --> 02:35:47,200 SO, IN TRADITIONAL HDS, YOU HAVE 3551 02:35:47,200 --> 02:35:54,640 YOUR ACCESS AND SO YOU WORK WITH 3552 02:35:54,640 --> 02:35:57,080 LIGANDS AND YOU LOW 3553 02:35:57,080 --> 02:35:57,960 CONCENTRATION OF PROTEIN. 3554 02:35:57,960 --> 02:36:04,240 SO YOU CAN EXPRESSION IT AND 3555 02:36:04,240 --> 02:36:05,040 THIS IS SWITCHED. 3556 02:36:05,040 --> 02:36:07,160 WE HAVE A HIGH CONCENTRATION OF 3557 02:36:07,160 --> 02:36:13,560 PROTEIN, USUALLY IN THE AND LOW 3558 02:36:13,560 --> 02:36:14,800 CONCENTRATION OF COMPOUNDS SO IF 3559 02:36:14,800 --> 02:36:17,160 YOU WANT TO HAVE THE BOUNDS YOU 3560 02:36:17,160 --> 02:36:19,680 COULD USE THE REST TO DO THAT. 3561 02:36:19,680 --> 02:36:23,400 NOW, IN PRACTICE, WE HAVE ONE 3562 02:36:23,400 --> 02:36:25,640 MILLION COPIES PAIR COMPOUNDS IN 3563 02:36:25,640 --> 02:36:26,200 OUR LIBRARY. 3564 02:36:26,200 --> 02:36:28,920 IF YOU WANT TO PUT THAT INTO 3565 02:36:28,920 --> 02:36:34,640 CONCENTRATION, THAT IS IN THE 3566 02:36:34,640 --> 02:36:37,600 MOLAR IMAGINE 10 TO THE MINUS 12 3567 02:36:37,600 --> 02:36:39,000 AND 10 TO THE MINUS 15. 3568 02:36:39,000 --> 02:36:41,480 SO WE HAVE A SMALL AMOUNT OF 3569 02:36:41,480 --> 02:36:42,440 COMPOUNDS IN OUR ASSAYS. 3570 02:36:42,440 --> 02:36:45,280 YOU CAN SEE IN DEL THE BINDING 3571 02:36:45,280 --> 02:36:47,600 IS DRIVEN BY PROTEIN 3572 02:36:47,600 --> 02:36:49,760 CONCENTRATION AND HDS THE 3573 02:36:49,760 --> 02:36:53,400 BINDING IS DRIVEN BY LIGAND 3574 02:36:53,400 --> 02:36:55,040 CONCENTRATION AND WE UTILIZE 3575 02:36:55,040 --> 02:36:58,520 THIS DURING THE SELECTION SO WE 3576 02:36:58,520 --> 02:37:00,160 ACTUALLY HAVE SAMPLES WHICH HAVE 3577 02:37:00,160 --> 02:37:02,480 HIGH PRE TEEN CONCENTRATION 3578 02:37:02,480 --> 02:37:04,880 AROUND TWO, TWO AND A HALF MACRO 3579 02:37:04,880 --> 02:37:06,240 MOLAR AND WE HAVE CONDITIONS 3580 02:37:06,240 --> 02:37:08,000 WHERE WE HAVE LOW PROTEIN 3581 02:37:08,000 --> 02:37:10,280 CONCENTRATIONS AND LET'S SAY .5 3582 02:37:10,280 --> 02:37:12,440 OR MICRO MORAL AND THIS GIVES US 3583 02:37:12,440 --> 02:37:14,600 AN IDEA OF THE RELATIVE AFFINITY 3584 02:37:14,600 --> 02:37:15,960 OF THE COMPOUNDS SO IF YOU HAVE 3585 02:37:15,960 --> 02:37:19,760 A COMPOUND THAT BINDS TO BOTH 3586 02:37:19,760 --> 02:37:22,280 SAMPLES THAN THE COMPOUND HAS 3587 02:37:22,280 --> 02:37:24,280 HIGHER AFFINITY. 3588 02:37:24,280 --> 02:37:26,960 IN A TYPICAL DELL SCREENING WE 3589 02:37:26,960 --> 02:37:30,720 OBSERVE BETWEEN 200 AND 500 FULL 3590 02:37:30,720 --> 02:37:31,640 ENRICHMENT PAIR ROUND OF 3591 02:37:31,640 --> 02:37:35,320 SELECTION SO THAT MEANS WE HAVE 3592 02:37:35,320 --> 02:37:37,800 TO RUN. 3593 02:37:37,800 --> 02:37:39,160 >>Alex: SELECTION TO REACH OUM 3594 02:37:39,160 --> 02:37:40,400 BOUNDS AND DRIVE THE BACKGROUND 3595 02:37:40,400 --> 02:37:42,840 DOWN SO HERE ON THE BOTTOM RIGHT 3596 02:37:42,840 --> 02:37:44,960 THERE'S JUST A SELECTION 3597 02:37:44,960 --> 02:37:47,800 STIMULATION AND ARE YOU SEE THE 3598 02:37:47,800 --> 02:37:50,560 ROUNDS OF DEL SELECTION FOR 3599 02:37:50,560 --> 02:37:52,240 COMPOUNDS WITH VARIOUS 3600 02:37:52,240 --> 02:37:55,120 AFFINITIES AND YOU CAN SEE IS 3601 02:37:55,120 --> 02:37:56,960 THAT THE COMPOUND THAT IS A 3602 02:37:56,960 --> 02:38:02,560 KNOWN BINDER SO IT HAS 100 MICRO 3603 02:38:02,560 --> 02:38:04,200 MOLARS IT'S WASHED OUT THE 3604 02:38:04,200 --> 02:38:05,200 SOLUTION AFTER THE SECOND ROUND 3605 02:38:05,200 --> 02:38:06,720 OF SELECTION SO AT THIS POINT, 3606 02:38:06,720 --> 02:38:10,400 WHAT YOU HAVE ARE COMPOUNDS WITH 3607 02:38:10,400 --> 02:38:12,880 10 MICRO MOLAR OR LOWER. 3608 02:38:12,880 --> 02:38:15,720 NOW, ESSENTIALLY, YOU COULD RUN 3609 02:38:15,720 --> 02:38:17,560 ENOUGH ROUNDS OF DEL SELECTION 3610 02:38:17,560 --> 02:38:19,640 TO REMOVE ALL OF YOUR LOWER 3611 02:38:19,640 --> 02:38:21,160 AFFINITY COMPOUNDS AND THE 3612 02:38:21,160 --> 02:38:22,840 REMAININGS ARE JUST THE HIGH 3613 02:38:22,840 --> 02:38:24,640 AFFINITY COMPOUNDS, HOWEVER IN 3614 02:38:24,640 --> 02:38:27,360 PRACTICE, WHAT WE DO IS WE LOWER 3615 02:38:27,360 --> 02:38:29,680 THE TOTAL NUMBER OF COMPOUNDS 3616 02:38:29,680 --> 02:38:31,400 JUST ENOUGH SO THAT WE CAN LOAD 3617 02:38:31,400 --> 02:38:33,640 THEM ONTO THE SEQUENCING CHIP 3618 02:38:33,640 --> 02:38:35,760 AND SO WE USUALLY RUN TWO OR 3619 02:38:35,760 --> 02:38:38,120 THREE ROUNDS OF SELECTION AND ON 3620 02:38:38,120 --> 02:38:39,840 UNDER THESE CONDITIONS, OUR HITS 3621 02:38:39,840 --> 02:38:43,120 HAVE EITHER SINGLE DIGIT MICRO 3622 02:38:43,120 --> 02:38:47,240 MOLAR AFFINITY SO NO MOLAR 3623 02:38:47,240 --> 02:38:50,160 AFFINITY SO THIS IS JUST AFTER 3624 02:38:50,160 --> 02:38:53,000 OVERVIEW OF THE VALIDATION AND 3625 02:38:53,000 --> 02:38:54,080 SCREENING PROCESS AND WE START 3626 02:38:54,080 --> 02:39:00,800 WITH A PRO SELECTION VALIDATION 3627 02:39:00,800 --> 02:39:11,320 AND AND WE RESYNTHESIZE ON DNA 3628 02:39:17,720 --> 02:39:19,800 TO CONFIRM THOSE HITS AND IF WE 3629 02:39:19,800 --> 02:39:21,520 CONFIRM THEY MAKE THEM OF DNA AS 3630 02:39:21,520 --> 02:39:24,160 WELL AND THEY'RE PUT INTO SOME 3631 02:39:24,160 --> 02:39:27,400 KIND OF BIOCHEMICAL AS SAY. 3632 02:39:27,400 --> 02:39:28,840 IT PROVED TO BE IMPORTANT IN OUR 3633 02:39:28,840 --> 02:39:30,120 WORK FLOW AND I WILL TALK ABOUT 3634 02:39:30,120 --> 02:39:33,280 THAT AS WELL A LITTLE BIT LATER. 3635 02:39:33,280 --> 02:39:35,560 AND LET ME START BY TELLING YOU 3636 02:39:35,560 --> 02:39:41,720 HOW WE VALIDATE OUR TARGETS AT 3637 02:39:41,720 --> 02:39:42,200 PFIZER. 3638 02:39:42,200 --> 02:39:45,040 EVERY SUCCESSFUL HEAT ID 3639 02:39:45,040 --> 02:39:47,040 PLATFORM DEPENDS ON THE REGIONS 3640 02:39:47,040 --> 02:39:48,840 AND THIS IS TRUE FOR DAL SINCE I 3641 02:39:48,840 --> 02:39:50,480 AS I MENTIONED, BINDING IS 3642 02:39:50,480 --> 02:39:52,240 DRIVEN BY PROTEIN CONCENTRATION 3643 02:39:52,240 --> 02:39:54,120 AND SO YOU REALLY WANT THE 3644 02:39:54,120 --> 02:39:58,280 MAJORITY OF YOUR PROTEINS TO BE 3645 02:39:58,280 --> 02:39:58,960 WELL BEHAVED. 3646 02:39:58,960 --> 02:40:03,880 SO, WITH THIS IN MIND, WE SET UP 3647 02:40:03,880 --> 02:40:05,400 A WORK FLOW BEFORE WE TAKE THEM 3648 02:40:05,400 --> 02:40:06,960 INTO THE SCREENING AND 3649 02:40:06,960 --> 02:40:08,080 ESSENTIALLY, WHAT WE DO OR TRY 3650 02:40:08,080 --> 02:40:10,760 TO DO IS TO QUANTIFY THE ACTIVE 3651 02:40:10,760 --> 02:40:14,160 SIDES OF THE PROTEIN AND BEFORE 3652 02:40:14,160 --> 02:40:21,080 I AM MOBILIZIMMOBILIZE, THE ACTN 3653 02:40:21,080 --> 02:40:23,360 BE LOST FOR SEVERAL REASONS, FOR 3654 02:40:23,360 --> 02:40:24,360 EXAMPLE, BECAUSE OF LOSS OF 3655 02:40:24,360 --> 02:40:26,480 TRANSITION OF FREEDOM IN THE 3656 02:40:26,480 --> 02:40:28,520 SPACE AND BECAUSE YOUR PROTEIN 3657 02:40:28,520 --> 02:40:30,920 REGION IS NOT JUST COMFORTABLE 3658 02:40:30,920 --> 02:40:35,400 WITH THE DEL REGIONS LIKE HIGH 3659 02:40:35,400 --> 02:40:37,160 SOLUBLE OR DNA. 3660 02:40:37,160 --> 02:40:38,480 THIS GRAPH I'M SHOWING YOU HERE 3661 02:40:38,480 --> 02:40:41,280 IS ONE OF THE OUR EARLIER TIMES 3662 02:40:41,280 --> 02:40:43,280 TO HAVE DATE A TARGET AND THIS 3663 02:40:43,280 --> 02:40:46,160 IS A FLUORESCENCE POLARIZATION 3664 02:40:46,160 --> 02:40:48,400 OF A COMPOUND BINDING TO THE 3665 02:40:48,400 --> 02:40:49,840 PROTEIN AND THIS IS PROTEIN IN 3666 02:40:49,840 --> 02:40:51,320 SOLUTIONS SO WITHOUT I AM 3667 02:40:51,320 --> 02:40:53,040 MOBILIZATION AND THIS IS HERE 3668 02:40:53,040 --> 02:40:54,880 WITH I AM MOBILIZATION AND NOW, 3669 02:40:54,880 --> 02:40:57,720 YOU CAN SEE THAT THE 3670 02:40:57,720 --> 02:40:59,120 PHARMACOLOGY MORE OR LESS REMAIN 3671 02:40:59,120 --> 02:41:01,120 THE SAME SO WE HAVE THE SAME 3672 02:41:01,120 --> 02:41:02,720 IC50 VALUE HOWEVER IF YOU LOOK 3673 02:41:02,720 --> 02:41:04,320 AT THE SIGNAL WINDOW IT'S 3674 02:41:04,320 --> 02:41:06,880 SIGNIFICANTLY DECREASES FOR THE 3675 02:41:06,880 --> 02:41:08,160 IMMOBILIZED PROTEIN. 3676 02:41:08,160 --> 02:41:11,120 WE'RE NOT SURE WHETHER THIS WAS 3677 02:41:11,120 --> 02:41:11,960 AN ARTIFACT OF THE ASSAYS OR 3678 02:41:11,960 --> 02:41:14,680 WHETHER MOST OF OUR PROTEINS 3679 02:41:14,680 --> 02:41:16,080 JUST DEMATURES AFTER 3680 02:41:16,080 --> 02:41:17,280 MOBILIZATION AND IT COULDN'T 3681 02:41:17,280 --> 02:41:18,800 BIND TO THE COMPOUND. 3682 02:41:18,800 --> 02:41:21,600 SO IT WAS CLEAR THAT THIS MATTER 3683 02:41:21,600 --> 02:41:24,880 IS NOT THE WAY FORWARD TO 3684 02:41:24,880 --> 02:41:28,200 VALIDATE OUR REGIONS FOR DEL 3685 02:41:28,200 --> 02:41:32,240 SINCE THIS IS NOT QUANTITIVE. 3686 02:41:32,240 --> 02:41:35,720 THE WORK WENT AHEAD AND SET UP A 3687 02:41:35,720 --> 02:41:36,920 WORKING FLOW AND IT'S CALLED 3688 02:41:36,920 --> 02:41:37,440 BELLY. 3689 02:41:37,440 --> 02:41:39,320 AND THE WORK FLOW IS A VERY 3690 02:41:39,320 --> 02:41:41,000 SIMILAR TO THE ACTUAL DEL WORK 3691 02:41:41,000 --> 02:41:43,200 FLOW SO YOU TAKE YOUR RECEPTORS 3692 02:41:43,200 --> 02:41:46,160 AND YOU IMMOBILIZE IT ON 3693 02:41:46,160 --> 02:41:50,800 AFFINITY BEADS AND AFTER I AM 3694 02:41:50,800 --> 02:41:53,240 MOBILIZATION YOU -- YOU SEPARATE 3695 02:41:53,240 --> 02:41:55,440 THE UNBOUND LIGAND AND THEN YOU 3696 02:41:55,440 --> 02:41:59,280 HAVE YOUR BOUNDING ON THE BEAT 3697 02:41:59,280 --> 02:42:00,960 FRACTION AND YOU USE BOTH OF 3698 02:42:00,960 --> 02:42:02,920 THESE FRACTION AND SOME KIND OF 3699 02:42:02,920 --> 02:42:10,200 DETECTION METHOD 3700 02:42:10,200 --> 02:42:11,000 THAT MEANS IT 3701 02:42:11,000 --> 02:42:12,400 DIDN'T BIND TO RECEPTOR. 3702 02:42:12,400 --> 02:42:15,560 HOWEVER, IF IT ENDED UP IN 3703 02:42:15,560 --> 02:42:17,480 THE BEAD OR FRACTION, THAT 3704 02:42:17,480 --> 02:42:18,840 INDICATE BINDING TO YOUR 3705 02:42:18,840 --> 02:42:22,040 PROTEIN. SO HERE ON THE TOP 3706 02:42:22,040 --> 02:42:24,520 LEFT, THIS IS A GRAPH USING 3707 02:42:24,520 --> 02:42:26,320 THE SAME PROTEIN 3708 02:42:26,320 --> 02:42:29,000 FLUORESCENCE LIGAND WE USED 3709 02:42:29,000 --> 02:42:31,040 BEFORE. WE USE ONE SINGLE 3710 02:42:31,040 --> 02:42:32,800 CONCENTRATION OF PROTEIN, 3711 02:42:32,800 --> 02:42:34,880 AND WE TITRATED IT WITH THE 3712 02:42:34,880 --> 02:42:37,560 TRACER. AFTER THAT WE 3713 02:42:37,560 --> 02:42:41,080 MEASURE FLUORESCENCE IN BOTH 3714 02:42:41,080 --> 02:42:44,560 THE ELUOTE AND SUPERNATANT 3715 02:42:44,560 --> 02:42:45,400 CONDITION. IN THESE 3716 02:42:45,400 --> 02:42:46,640 CONDITIONS THE BINDING 3717 02:42:46,640 --> 02:42:48,480 SHOULD BE STOIC METRIC AND 3718 02:42:48,480 --> 02:42:50,480 THE FLUORESCENT TRACER 3719 02:42:50,480 --> 02:42:54,040 NICELY SHOWED UP IN THE 3720 02:42:54,040 --> 02:42:55,760 ELUOTE FRACTION AS EXPECTED 3721 02:42:55,760 --> 02:42:57,440 AND THAT WAS INFLECTION 3722 02:42:57,440 --> 02:42:59,400 POINT OF 500 NANOMOLAR WHICH 3723 02:42:59,400 --> 02:43:00,880 IS CONCENTRATION OF PROTEIN 3724 02:43:00,880 --> 02:43:02,800 WE PUT INTO THE ASSAY. SO 3725 02:43:02,800 --> 02:43:05,240 THIS IS A VERY NICE DATA, WE 3726 02:43:05,240 --> 02:43:07,240 WERE HAPPY TO SEE THIS. 3727 02:43:07,240 --> 02:43:08,160 HOWEVER IF YOU THINK ABOUT 3728 02:43:08,160 --> 02:43:09,880 IT FOR MOST OF YOUR TARGET 3729 02:43:09,880 --> 02:43:11,400 FOR MOST OF YOUR PROJECT, 3730 02:43:11,400 --> 02:43:13,080 YOU ARE NOT GOING TO HAVE A 3731 02:43:13,080 --> 02:43:15,800 FLUORESCENT LIGAND. IT WOULD 3732 02:43:15,800 --> 02:43:17,720 TAKE JUST TOO MUCH TIME AND 3733 02:43:17,720 --> 02:43:19,840 EFFORT TO SYNTHESIZE 3734 02:43:19,840 --> 02:43:21,680 FLUORESCENT LIGAND. TO 3735 02:43:21,680 --> 02:43:23,960 ADDRESS THIS, THE GROUP 3736 02:43:23,960 --> 02:43:25,320 QUESTIONED WHETHER WE COULD 3737 02:43:25,320 --> 02:43:27,200 USE MASS SPEC AS DETECTION 3738 02:43:27,200 --> 02:43:29,000 METHOD TOWARD THIS BALI WORK 3739 02:43:29,000 --> 02:43:30,320 FLOW. SO WE REPEATED THE 3740 02:43:30,320 --> 02:43:32,360 ASSAY USING THE SAME 3741 02:43:32,360 --> 02:43:35,760 PROTEIN, BUT THE STANDARD 3742 02:43:35,760 --> 02:43:38,600 LIGAND DIDN'T VICE PRESIDENT 3743 02:43:38,600 --> 02:43:41,200 LIGAND ON IT, SEPARATED AND 3744 02:43:41,200 --> 02:43:43,040 SEPARATED THE COMPOUND USING 3745 02:43:43,040 --> 02:43:45,920 MASS SPMASS SPEC. YOU CAN SEE 3746 02:43:45,920 --> 02:43:46,880 ACCUMULATION OF COMPOUND 3747 02:43:46,880 --> 02:43:49,080 WITH INFLECTION POINT AROUND 3748 02:43:49,080 --> 02:43:49,920 750 NANOMOLAR. THE 3749 02:43:49,920 --> 02:43:51,600 INFLECTION POINT BETWEEN THE 3750 02:43:51,600 --> 02:43:53,120 TWO EXPERIMENT SHIFT AD 3751 02:43:53,120 --> 02:43:54,400 LITTLE BIT SIMPLY BECAUSE WE 3752 02:43:54,400 --> 02:43:57,400 PUT MORE PROTEIN INTO THE 3753 02:43:57,400 --> 02:44:00,920 BALI MS EXPERIMENT. SO THIS 3754 02:44:00,920 --> 02:44:03,200 BALI MS BECAME THE CORNER E 3755 02:44:03,200 --> 02:44:04,040 STONE OF REGION VALIDATION 3756 02:44:04,040 --> 02:44:06,000 AT PFIZER, IT IS VERY QUICK 3757 02:44:06,000 --> 02:44:09,640 TO RUN, IF YOUR PROTEIN IS 3758 02:44:09,640 --> 02:44:10,880 MAL BEHAVED WE CAN VALIDATE 3759 02:44:10,880 --> 02:44:15,800 WITHIN A WEEK OR TWO. SO YOU 3760 02:44:15,800 --> 02:44:18,560 VALIDATED YOUR REAGENT, YOU 3761 02:44:18,560 --> 02:44:20,120 RAN YOUR SCREEN AND AT THE 3762 02:44:20,120 --> 02:44:21,920 END YOU HAVE NO HITS. OR YOU 3763 02:44:21,920 --> 02:44:23,720 HAVE HITS WITH POOR PHYSICAL 3764 02:44:23,720 --> 02:44:25,320 CHEMICAL PROPERTIES OR YOU 3765 02:44:25,320 --> 02:44:27,480 MAKE YOUR HIT OF DNA AND 3766 02:44:27,480 --> 02:44:28,520 THEY JUST CONFIRM. SO WHAT 3767 02:44:28,520 --> 02:44:30,760 DO YOU DO NEXT? TO ADDRESS 3768 02:44:30,760 --> 02:44:33,040 ALL OF THESE QUESTIONS, THE 3769 02:44:33,040 --> 02:44:34,640 PFIZER GROUP SET UP A WORK 3770 02:44:34,640 --> 02:44:36,800 FLOW, BUT BEFORE I GET INTO 3771 02:44:36,800 --> 02:44:39,240 THE DETAILS LET ME SHOW YOU 3772 02:44:39,240 --> 02:44:40,400 HOW WIAL VISUALIZE AND 3773 02:44:40,400 --> 02:44:41,440 ANALYZE THE DATA SINCE I'M 3774 02:44:41,440 --> 02:44:43,960 GOING TO BE SHOWING THESE 3775 02:44:43,960 --> 02:44:45,440 PLOT BY PLOT. SO THIS IS A 3776 02:44:45,440 --> 02:44:47,440 3D PLOT WE CALL THIS THE 3777 02:44:47,440 --> 02:44:49,960 CUBE. AND ON THE ACCESS OF 3778 02:44:49,960 --> 02:44:51,280 THE CUBE YOU HAVE YOUR 3779 02:44:51,280 --> 02:44:52,320 BUILDING BLOCKS YOUR 3780 02:44:52,320 --> 02:44:54,920 MONOMERS. SO THIS MEANS THAT 3781 02:44:54,920 --> 02:44:56,720 EVERY SINGLE COMPOUND IN 3782 02:44:56,720 --> 02:44:58,120 YOUR LIBRARY, WILL HAVE A 3783 02:44:58,120 --> 02:44:59,800 UNIQUE PHYSICAL ADDRESS IN 3784 02:44:59,800 --> 02:45:03,240 THE 3D SPAC SPACE. SO THESE 3785 02:45:03,240 --> 02:45:04,560 SPHERES YOU SEE HERE ARE 3786 02:45:04,560 --> 02:45:06,600 COMPOUNDS. AND THEY ARE 3787 02:45:06,600 --> 02:45:07,960 SIZED BY THE READING COUNT 3788 02:45:07,960 --> 02:45:09,960 WE GOT OUT OF NEXT 3789 02:45:09,960 --> 02:45:12,400 GENERATION SEQUENCING. NOW 3790 02:45:12,400 --> 02:45:14,680 WHEN MEDICINAL CHEMIST LOOKS 3791 02:45:14,680 --> 02:45:17,400 AT THIS DATA THEY LOOK FOR 3792 02:45:17,400 --> 02:45:19,040 FEATURES LIKE LINE FEATURE 3793 02:45:19,040 --> 02:45:20,640 OR A PLANE FEATURE. SO FOR 3794 02:45:20,640 --> 02:45:24,720 EXAMPLE THIS LINE FEATURE IS 3795 02:45:24,720 --> 02:45:26,080 MEDICINAL CHEMIST IS 3796 02:45:26,080 --> 02:45:27,600 INTERESTED IN. THESE 3797 02:45:27,600 --> 02:45:29,120 FEATURES CAN OCCUR FOR 3798 02:45:29,120 --> 02:45:31,760 VARIOUS REASON. IT CAN BE IN 3799 02:45:31,760 --> 02:45:34,360 THIS CASE IT CAN BE STR 3800 02:45:34,360 --> 02:45:36,480 AROUND THIS PINK BUILDING 3801 02:45:36,480 --> 02:45:37,960 BLOCK OR IT ALSO THE 3802 02:45:37,960 --> 02:45:39,160 POSSIBLE THE BUILDING BLOCK 3803 02:45:39,160 --> 02:45:40,400 THE PINK BUILDING BLOCK IS 3804 02:45:40,400 --> 02:45:41,920 NOT IN THE BINDING POCKET SO 3805 02:45:41,920 --> 02:45:42,880 IT DOES NOT CONTRIBUTE TO 3806 02:45:42,880 --> 02:45:46,280 BINDING. OR THE ASSERTION 3807 02:45:46,280 --> 02:45:48,840 IS MAYBE BECAUSE OF FAIR 3808 02:45:48,840 --> 02:45:50,800 CHEMISTRY OR TRUNCATIONS AND 3809 02:45:50,800 --> 02:45:51,640 I WILL MENTION THAT A LITTLE 3810 02:45:51,640 --> 02:45:56,160 BIT LATER AS WELL. SO AS I 3811 02:45:56,160 --> 02:45:57,840 SAID AT THE BEGINNING, AT 3812 02:45:57,840 --> 02:46:00,480 THE END OF OUR SCREEN IF YOU 3813 02:46:00,480 --> 02:46:01,480 WANT TO END UP WITH 3814 02:46:01,480 --> 02:46:03,320 COMPOUNDS WITH GOOD PHYSICAL 3815 02:46:03,320 --> 02:46:05,240 CHEMICAL PROPERTIES, DRUG 3816 02:46:05,240 --> 02:46:06,880 LIKE PROPERTIES SOMETHING 3817 02:46:06,880 --> 02:46:08,480 MEDICINAL CHEMIST CAN 3818 02:46:08,480 --> 02:46:09,520 CONVERT TO FURTHER DEVELOP 3819 02:46:09,520 --> 02:46:11,120 AND DO ASSAY ALL AROUND. NOW 3820 02:46:11,120 --> 02:46:12,920 A LOT OF THINGS THAT 3821 02:46:12,920 --> 02:46:14,480 ATTRIBUTES TO THESE -- A LOT 3822 02:46:14,480 --> 02:46:16,000 OF PROPERTIES THAT 3823 02:46:16,000 --> 02:46:17,840 ATTRIBUTES TO THIS, 3824 02:46:17,840 --> 02:46:21,560 MOLECULAR WEIGHT, HYDROGEN 3825 02:46:21,560 --> 02:46:23,240 COUNTS, AUTOMATIC RINGS C 3826 02:46:23,240 --> 02:46:25,440 LOG D, SO ON. SO WE 3827 02:46:25,440 --> 02:46:28,240 ESTABLISHED AN MPO SCORING 3828 02:46:28,240 --> 02:46:30,080 SYSTEM TO GUIDE OUR HIT 3829 02:46:30,080 --> 02:46:33,680 SELECTION DURING ANALYSIS. 3830 02:46:33,680 --> 02:46:34,560 THIS SCORING SYSTEM RELIES 3831 02:46:34,560 --> 02:46:38,240 ON MOLECULAR WEIGHT M 3832 02:46:38,240 --> 02:46:40,240 HYDROGEN BOND, AUTOMATIC 3833 02:46:40,240 --> 02:46:42,520 RING COUNTS, AND USE 3834 02:46:42,520 --> 02:46:43,960 VISUALIZATION TOOLS TO HELP 3835 02:46:43,960 --> 02:46:45,040 THE SELECTION. SO THIS IS 3836 02:46:45,040 --> 02:46:46,440 JUST AN EXAMPLE AT THE 3837 02:46:46,440 --> 02:46:49,000 BOTTOM. SO THIS LINE 3838 02:46:49,000 --> 02:46:51,000 FEATURE, WAS A HIT IN ONE OF 3839 02:46:51,000 --> 02:46:52,400 OUR SELECTION. AND YOU CAN 3840 02:46:52,400 --> 02:46:54,560 SEE THAT THE BIGGEST 3841 02:46:54,560 --> 02:46:56,000 SPHERES, THE ONES THAT HAVE 3842 02:46:56,000 --> 02:46:57,320 THE MOST COUNTS, THEY ARE 3843 02:46:57,320 --> 02:46:59,000 ACTUALLY COLORED RED AND 3844 02:46:59,000 --> 02:47:00,000 ORANGE AND THAT MEANS THAT 3845 02:47:00,000 --> 02:47:02,440 THEY HAVE A LOW MPO SCORE, 3846 02:47:02,440 --> 02:47:03,600 SO THEY HAVE VERY POOR 3847 02:47:03,600 --> 02:47:05,200 PHYSICAL CHEMICAL 3848 02:47:05,200 --> 02:47:07,240 PROPERTIES. HOWEVER, ONCE WE 3849 02:47:07,240 --> 02:47:09,400 APPLY MPO SCORE BASE 3850 02:47:09,400 --> 02:47:11,720 FILTERING WE REMAIN THESE 3851 02:47:11,720 --> 02:47:12,640 GREENISH COLOR SPHERES, 3852 02:47:12,640 --> 02:47:14,640 THESE ARE COMPOUNDS THAT 3853 02:47:14,640 --> 02:47:16,560 HAVE BETTER PHYSICAL 3854 02:47:16,560 --> 02:47:17,400 CHEMICAL PROPERTIES 3855 02:47:17,400 --> 02:47:18,000 SOMETHING YOU WANTS TO WORK 3856 02:47:18,000 --> 02:47:20,360 WITH T AT THE END. NOW, I 3857 02:47:20,360 --> 02:47:22,680 SHOWED THIS FILTERING JUST 3858 02:47:22,680 --> 02:47:24,200 FOR DEMONSTRATION PURPOSES, 3859 02:47:24,200 --> 02:47:26,120 IN REALITY WE DON'T USE THE 3860 02:47:26,120 --> 02:47:28,080 MPO SCORING TO FILTER OUT 3861 02:47:28,080 --> 02:47:30,960 DATA, WE JUST USE IT AS A 3862 02:47:30,960 --> 02:47:33,200 TOOL TO GUIDE HIT 3863 02:47:33,200 --> 02:47:36,800 SELECTIONS. OKAY. SO AT THE 3864 02:47:36,800 --> 02:47:37,880 BEGINNING WHEN THE GROUP 3865 02:47:37,880 --> 02:47:39,240 JUST STARTED A COUPLE OF 3866 02:47:39,240 --> 02:47:42,360 YEARS AGO, THE -- OUR 3867 02:47:42,360 --> 02:47:44,240 SCREENING PROCESS CONSISTED 3868 02:47:44,240 --> 02:47:46,520 OF DOING THE SELECTION, 3869 02:47:46,520 --> 02:47:48,320 ANALYZE THE DATA, CALLING 3870 02:47:48,320 --> 02:47:50,120 THE HITS, THEN RIGHT AWAY 3871 02:47:50,120 --> 02:47:51,600 SYNTHESIZE THE COMPOUNDS OF 3872 02:47:51,600 --> 02:47:52,960 DNA AND PUT THEM INTO SOME 3873 02:47:52,960 --> 02:47:54,560 KIND OF FUNCTIONALIZED 3874 02:47:54,560 --> 02:47:58,120 BINDING ASSAY. WHAT WE SAW 3875 02:47:58,120 --> 02:48:00,200 IS THAT AFTER ALL THAT 3876 02:48:00,200 --> 02:48:01,360 EFFORT AND RESOURCES THAT WE 3877 02:48:01,360 --> 02:48:03,840 PUT INTO MAKING THE 3878 02:48:03,840 --> 02:48:05,760 COMPOUNDS OF DNA, MOST OF 3879 02:48:05,760 --> 02:48:07,000 THEM DIDN'T CONFIRM. AND 3880 02:48:07,000 --> 02:48:08,960 THEY REALLY HAD A -- WE HAD 3881 02:48:08,960 --> 02:48:10,480 A LOW CONFIRMATION RATE. SO 3882 02:48:10,480 --> 02:48:13,880 NOW HOW IS THIS POSSIBLE 3883 02:48:13,880 --> 02:48:15,480 THAT BALI INDICATED THE 3884 02:48:15,480 --> 02:48:17,320 COMPOUND IS A HIT AND MAKE 3885 02:48:17,320 --> 02:48:19,720 IT OF DNA AND IT DOESN'T 3886 02:48:19,720 --> 02:48:22,360 CONFIRM? REMEMBER AT THE 3887 02:48:22,360 --> 02:48:25,360 BEGINNING I SAID EVERY DNA 3888 02:48:25,360 --> 02:48:27,680 TAG WAS ONE COMPOUND. BUT I 3889 02:48:27,680 --> 02:48:30,600 LIED. IN REALITY, WHAT 3890 02:48:30,600 --> 02:48:32,400 HAPPENS IS THAT DNA LIGATION 3891 02:48:32,400 --> 02:48:33,880 OCCURS WITH HIGH FIDELITY AS 3892 02:48:33,880 --> 02:48:35,960 YOU WOULD EXPECT IT. 3893 02:48:35,960 --> 02:48:36,640 HOWEVER, THIS IS NOT TRUE 3894 02:48:36,640 --> 02:48:38,640 FOR CHEMICAL SYNTHESIS. 3895 02:48:38,640 --> 02:48:39,840 ESPECIALLY UNDER CONDITIONS 3896 02:48:39,840 --> 02:48:41,720 THAT WE USE, TO PREVENT DNA 3897 02:48:41,720 --> 02:48:44,280 DAMAGE. SO FOR EXAMPLE IN 3898 02:48:44,280 --> 02:48:45,800 THREE CYCLE CHEMISTRY, IF 3899 02:48:45,800 --> 02:48:48,040 THIS IS YOUR TARGET 3900 02:48:48,040 --> 02:48:49,120 COMPOUND, IT IS POSSIBLE 3901 02:48:49,120 --> 02:48:52,040 SOMETHING GOES BAD DURING 3902 02:48:52,040 --> 02:48:56,480 CYCLE 1, AND SAY THE NYTRIL 3903 02:48:56,480 --> 02:48:57,680 GROUP IS NOT REDUCED TO 3904 02:48:57,680 --> 02:48:59,440 AMINE OR FIRST BLOCK IS NOT 3905 02:48:59,440 --> 02:49:00,360 INCORPORATED. 3906 02:49:00,360 --> 02:49:03,120 FOR THIS ONE FINAL COMPOUND, 3907 02:49:03,120 --> 02:49:04,880 POTENTIALLY YOU COULD GET ALL OF 3908 02:49:04,880 --> 02:49:06,600 THESE CYCLE DUCTS YOU CAN SEE ON 3909 02:49:06,600 --> 02:49:10,360 THE SCREEN, AND IT FORMED ALL 3910 02:49:10,360 --> 02:49:11,800 THE CENTER DUCTS HAVE THE SAME 3911 02:49:11,800 --> 02:49:15,400 DNA TAG. SO THE ONE DNA CODE 3912 02:49:15,400 --> 02:49:17,280 EQUALS ONE SMALL MOLECULE 3913 02:49:17,280 --> 02:49:19,960 STATEMENT IS NOT TRUE. AND THIS 3914 02:49:19,960 --> 02:49:22,520 FAILED CHEMISTRY AND TRUNCATED 3915 02:49:22,520 --> 02:49:24,320 STRUCTURES NEEDS TO BE TAKEN 3916 02:49:24,320 --> 02:49:27,000 ACCOUNT DURING DATA ANALYSIS. 3917 02:49:27,000 --> 02:49:28,720 BUT EXPERIENCE MEDICINAL 3918 02:49:28,720 --> 02:49:32,080 SCIENTIST CAN ACTUALLY DO THAT. 3919 02:49:32,080 --> 02:49:36,160 OKAY. SO NOW WE KNOW THAT WE 3920 02:49:36,160 --> 02:49:38,720 HAVE PROBABLY ALL THESE TYPE OF 3921 02:49:38,720 --> 02:49:40,600 REACTION MIXTURE, SO WE HAD TO 3922 02:49:40,600 --> 02:49:42,840 SET UP A WORK FLOW TO ADDRESS 3923 02:49:42,840 --> 02:49:44,320 OUR LOW CONFIRMATION RATE. AND 3924 02:49:44,320 --> 02:49:46,680 I'M GOING TO USE THIS COMPOUND 3925 02:49:46,680 --> 02:49:50,600 BOND AS AN EXAMPLE. SO FROM THIS 3926 02:49:50,600 --> 02:49:53,720 SLIDE FEATURE COMPOUND ONE WAS 3927 02:49:53,720 --> 02:49:56,040 CALLED AS A HIT BECAUSE OF GOOD 3928 02:49:56,040 --> 02:49:57,520 PHYSICAL CHEMICAL PROPERTIES 3929 02:49:57,520 --> 02:50:00,120 THAT IT HAS. HOWEVER, WE MADE 3930 02:50:00,120 --> 02:50:03,080 THEM OF DNA AND IT JUST DIDN'T 3931 02:50:03,080 --> 02:50:05,640 CONFIRM. SO THE GROUP WENT AND 3932 02:50:05,640 --> 02:50:07,600 RESYNTHESIZED THIS COMPOUND ON 3933 02:50:07,600 --> 02:50:09,880 THE DNA HEADPIECE,S USING THE 3934 02:50:09,880 --> 02:50:11,920 EXACT SAME CHEMICAL RULE THAT 3935 02:50:11,920 --> 02:50:14,000 WAS USED DURING THE ORIGINAL 3936 02:50:14,000 --> 02:50:17,240 SYNTHESIS. SO AFTER 3937 02:50:17,240 --> 02:50:18,720 RESYNTHESIZING THIS, THEY 3938 02:50:18,720 --> 02:50:20,520 ANALYZED THE COMPOUND MIXTURE, 3939 02:50:20,520 --> 02:50:22,880 AND THIS IS JUST THE HPLC TRACE 3940 02:50:22,880 --> 02:50:25,200 FOR THAT MIXTURE. AND 3941 02:50:25,200 --> 02:50:26,720 SURPRISINGLY COMPOUND ONE WASN'T 3942 02:50:26,720 --> 02:50:30,120 EVEN THERE. INSTEAD, WE HAD TWO 3943 02:50:30,120 --> 02:50:32,440 MAJOR CYCLE DUCTS WHERE THE 3944 02:50:32,440 --> 02:50:34,720 FIRST BUILDING BLOCK WASN'T 3945 02:50:34,720 --> 02:50:35,680 INCORPORATED, OR THE SECOND 3946 02:50:35,680 --> 02:50:37,280 BUILDING BLOCK WAS INCORPORATED 3947 02:50:37,280 --> 02:50:40,600 TWICE. SO WE ENDED UP WITH THIS 3948 02:50:40,600 --> 02:50:43,400 COMPOUND 2 AND COMPOUND 3, WHICH 3949 02:50:43,400 --> 02:50:45,440 HAD REVERSE CHEMICAL PHYSICAL 3950 02:50:45,440 --> 02:50:47,880 PROPERTY -- WORSE PROPERTIES 3951 02:50:47,880 --> 02:50:49,120 THAN COMPOUND ONE. THIS IS 3952 02:50:49,120 --> 02:50:52,320 GREAT, WE KNOW WHAT WAS IN OUR 3953 02:50:52,320 --> 02:50:53,480 ORIGINAL SELECTION BUT THIS 3954 02:50:53,480 --> 02:50:55,360 STILL DOESN'T GIVE THE ANSWER 3955 02:50:55,360 --> 02:50:58,320 WHICH COMPOUND BOUND TO PROTEIN. 3956 02:50:58,320 --> 02:51:00,680 SO WE WENT AHEAD AND UTILIZED 3957 02:51:00,680 --> 02:51:04,480 THIS BALI MS PET PLATFORM FOR 3958 02:51:04,480 --> 02:51:07,000 DNA CONFIRMATION AS WELL. SO AS 3959 02:51:07,000 --> 02:51:10,760 BEFORE, WE IMMOBILIZE OUR 3960 02:51:10,760 --> 02:51:12,760 RECEPTOR UNDER BEADS THEN 3961 02:51:12,760 --> 02:51:14,640 INCUBATED WITH THE OWN DNA 3962 02:51:14,640 --> 02:51:17,200 COMPOUND MIXTURE. AFTER 3963 02:51:17,200 --> 02:51:19,400 REACHING REWILLIE YUM WE 3964 02:51:19,400 --> 02:51:21,000 SEPARATE SUPERNATANT FROM BEADS 3965 02:51:21,000 --> 02:51:25,040 AND INJECT INTO LCMS. DURING 3966 02:51:25,040 --> 02:51:26,600 INJECTION YOU USE INTERNAL 3967 02:51:26,600 --> 02:51:29,240 SENDER SO THAT ALLOWS US TO 3968 02:51:29,240 --> 02:51:31,480 QUANTIFY TO NORMALIZE OUR DATA 3969 02:51:31,480 --> 02:51:33,240 AND ACCESS THE DATA 3970 02:51:33,240 --> 02:51:36,000 QUANTITATIVELY. SO BASED ON THE 3971 02:51:36,000 --> 02:51:38,680 LCMS DATA, WE SAW A FIVE FOLD 3972 02:51:38,680 --> 02:51:43,280 ENRICHMENT FOR COMPOUND 2 AND 3973 02:51:43,280 --> 02:51:46,160 SURFACE 3, 4, INRICHMENT FOR 3974 02:51:46,160 --> 02:51:47,160 COMPOUND 3 SO IT IS HIGHLY 3975 02:51:47,160 --> 02:51:49,520 LIKELY THE ORIGINAL WAS THE SUM 3976 02:51:49,520 --> 02:51:54,160 OF BOTH OF THESE CONTRIBUTIONS. 3977 02:51:54,160 --> 02:51:57,360 SO NOW, INSTEAD OF GOING 3978 02:51:57,360 --> 02:52:00,040 STRAIGHT TO OFF DNA SYNTHESIS, 3979 02:52:00,040 --> 02:52:02,280 FIRST WE RESYNTHESIZED OUR DEL 3980 02:52:02,280 --> 02:52:04,920 HITS ON DNA TO GET THIS HIT TREE 3981 02:52:04,920 --> 02:52:06,640 THEN ANALYZE THAT MIXTURE USING 3982 02:52:06,640 --> 02:52:10,680 OUR BALI MS PLATFORM. THIS THOSE 3983 02:52:10,680 --> 02:52:12,880 COMPOUNDS -- IF THOSE COMPOUNDS 3984 02:52:12,880 --> 02:52:14,560 CONFIRM WE MAKE THEM OF DNA AS 3985 02:52:14,560 --> 02:52:15,720 WELL AND PUT THEM INTO A 3986 02:52:15,720 --> 02:52:19,240 FUNCTIONAL ASSAY. SO WITH THIS 3987 02:52:19,240 --> 02:52:22,200 APPROACH WE GOT HIGHER 3988 02:52:22,200 --> 02:52:23,640 CONFIRMATION RATE AND THIS ALSO 3989 02:52:23,640 --> 02:52:25,760 ALLOWED US TO FOLLOW-UP ON MORE 3990 02:52:25,760 --> 02:52:28,840 CHEMICAL SERIES AND MAKE FEWER 3991 02:52:28,840 --> 02:52:30,960 OF DNA COMPOUNDS SO ESSENTIALLY 3992 02:52:30,960 --> 02:52:35,440 SAVING THESE SOURCES FOR THE 3993 02:52:35,440 --> 02:52:36,840 PROAPOLOGETIC. NOW THAT YOU KNOW 3994 02:52:36,840 --> 02:52:38,360 ALL ABOUT DEL, I WANTED TO SHOW 3995 02:52:38,360 --> 02:52:40,920 YOU AN EXAMPLE OF SELECTION 3996 02:52:40,920 --> 02:52:45,000 PLAN. AS I MENTIONED BEFORE, WE 3997 02:52:45,000 --> 02:52:46,800 CAN PENALIZE CONDITIONS DURING 3998 02:52:46,800 --> 02:52:48,760 DEL SELECTION. SO THAT MEANS 3999 02:52:48,760 --> 02:52:51,160 THAT YOU CAN HAVE YOUR FULL 4000 02:52:51,160 --> 02:52:52,680 PROTEIN FOR EXAMPLE IN ONE 4001 02:52:52,680 --> 02:52:53,800 CONDITION AND IN OTHER CONDITION 4002 02:52:53,800 --> 02:52:55,560 MAYBE YOU HAVE THE PROTEIN IN 4003 02:52:55,560 --> 02:52:56,600 COMPLEX OR MAYBE YOU HAVE THE 4004 02:52:56,600 --> 02:52:59,720 MUTANT PROTEIN. BUT IT'S ALSO 4005 02:52:59,720 --> 02:53:01,480 POSSIBLE YOU WANT TO ADD 4006 02:53:01,480 --> 02:53:03,240 SUPPLEMENTS LIKE CHAPERONE OR 4007 02:53:03,240 --> 02:53:05,040 CO-FACTOR OR SOME SUBSTRATE AS 4008 02:53:05,040 --> 02:53:07,400 WELL TO SEE COMPETITION WITH THE 4009 02:53:07,400 --> 02:53:08,640 SUBSTRATE. I MENTIONED BEFORE 4010 02:53:08,640 --> 02:53:11,080 THAT WE USUALLY RUN TWO 4011 02:53:11,080 --> 02:53:13,120 CONCENTRATIONS, HIGHER AND LOWER 4012 02:53:13,120 --> 02:53:15,320 CONCENTRATION, TO GET THE IDEA 4013 02:53:15,320 --> 02:53:18,240 OF THE RELATIVE AFFINITY OF THE 4014 02:53:18,240 --> 02:53:19,520 COMPOUNDS. I DIDN'T MENTION 4015 02:53:19,520 --> 02:53:22,560 REALLY THIS, BUT WE TRY TO ALSO 4016 02:53:22,560 --> 02:53:23,960 INCLUDE A POSITIVE CONTROL IN 4017 02:53:23,960 --> 02:53:27,120 OUR ASSAY. SO IF THE PROTEIN OF 4018 02:53:27,120 --> 02:53:28,680 INTEREST HAS SOME KIND OF 4019 02:53:28,680 --> 02:53:30,400 CHEMICAL MATTER, THAT BINDS TO 4020 02:53:30,400 --> 02:53:33,480 THE PROTEIN WITH RELATIVELY HIGH 4021 02:53:33,480 --> 02:53:35,440 AFFINITY, WE CAN MAKE THAT 4022 02:53:35,440 --> 02:53:37,120 COMPOUND ON THE DNA SPIKE IT 4023 02:53:37,120 --> 02:53:40,160 INTO THE DNA DEL POOL, AND THEN 4024 02:53:40,160 --> 02:53:43,280 USE IT DURING SELECTION. SO 4025 02:53:43,280 --> 02:53:44,720 THAT'S HIGH COMPOUND WILL SERVE 4026 02:53:44,720 --> 02:53:47,160 AS POSITIVE CONTROL, AND WE CAN 4027 02:53:47,160 --> 02:53:48,760 MONITOR IT DURING SELECTION AND 4028 02:53:48,760 --> 02:53:51,280 SEE IF IT WAS ENRICHED. SO FOR 4029 02:53:51,280 --> 02:53:52,920 EXAMPLE, IF AT THE END OF THE 4030 02:53:52,920 --> 02:53:55,080 DAY YOU DON'T GET ANY HIT OUT OF 4031 02:53:55,080 --> 02:53:57,560 YOUR DEL SELECTION BUT YOUR 4032 02:53:57,560 --> 02:53:59,320 POSITIVE CONTROL WAS ENRICHED, 4033 02:53:59,320 --> 02:54:02,400 YOU CAN BE SURE THAT THE DEL 4034 02:54:02,400 --> 02:54:03,960 SELECTION WENT WELL JUST 4035 02:54:03,960 --> 02:54:06,200 SOMETHING ELSE HAPPENING THAT 4036 02:54:06,200 --> 02:54:08,440 DIDN'T GIVE YOU ANY HITS. THIS 4037 02:54:08,440 --> 02:54:09,600 SMALL TABLE IS TO GIVE YOU AN 4038 02:54:09,600 --> 02:54:11,360 IDEA OF THE SELECTION PLAN, SO 4039 02:54:11,360 --> 02:54:13,800 FOR EXAMPLE IN SAMPLE ONE AND 4040 02:54:13,800 --> 02:54:16,280 TWO, WE HAVE HIGH PROTEIN 4041 02:54:16,280 --> 02:54:18,040 CONCENTRATION, FOR SAMPLE TWO WE 4042 02:54:18,040 --> 02:54:20,400 ADDED A NON-BINDER AS WELL TO 4043 02:54:20,400 --> 02:54:22,320 IDENTIFY DEL COMPOUNDS THAT 4044 02:54:22,320 --> 02:54:23,320 COULD COMPETE WITH THIS 4045 02:54:23,320 --> 02:54:24,520 NON-BINDER. SO ESSENTIALLY BIND 4046 02:54:24,520 --> 02:54:28,280 TO THE SAME BINDING POCKET. AND 4047 02:54:28,280 --> 02:54:30,560 THEN IN FOUR SAMPLES 3 AND 4 WE 4048 02:54:30,560 --> 02:54:31,760 HAVE THE LOW PROTEIN 4049 02:54:31,760 --> 02:54:33,760 CONCENTRATION AND AGAIN FOR 4 WE 4050 02:54:33,760 --> 02:54:36,600 ADDED A NON-BINDER. FOR SAMPLE 5 4051 02:54:36,600 --> 02:54:38,840 THAT'S BASICALLY YOUR NEGATIVE 4052 02:54:38,840 --> 02:54:40,400 CONTROL, YOU DON'T ADD ANY 4053 02:54:40,400 --> 02:54:42,040 PROTEIN AND YOU JUST WORK WITH 4054 02:54:42,040 --> 02:54:44,080 THE BEADS TO IDENTIFY BINDERS 4055 02:54:44,080 --> 02:54:46,360 THAT ARE BEING CAPTURED BY 4056 02:54:46,360 --> 02:54:49,840 MAGNETIC BEADS. SO I THINK I 4057 02:54:49,840 --> 02:54:53,280 HAVE SOME TIME LEFT TO MENTION 4058 02:54:53,280 --> 02:54:56,000 CELL BASED SCREENING AND THUS 4059 02:54:56,000 --> 02:54:58,040 SCREENING FOR MEMBRANE PROTEINS 4060 02:54:58,040 --> 02:54:59,400 SO WHY ARE WE INTERESTED IN 4061 02:54:59,400 --> 02:55:00,880 MEMBRANE PROTEIN? 4062 02:55:00,880 --> 02:55:02,760 BECAUSE THEY PERFORM A MYRIAD OF 4063 02:55:02,760 --> 02:55:05,040 BIOLOGICAL FUNCTIONS AND THEY 4064 02:55:05,040 --> 02:55:08,560 ARE IMPORTANT DRUG TARGETS. SO 4065 02:55:08,560 --> 02:55:11,000 THIS GRAPH IS SHOWS THE FD 4066 02:55:11,000 --> 02:55:12,720 APPROVED SMALL MOLECULAR DRUGS 4067 02:55:12,720 --> 02:55:15,000 TARGETING MAJOR PROTEIN 4068 02:55:15,000 --> 02:55:18,000 FAMILIES. IT IS FROM 2017 SO 4069 02:55:18,000 --> 02:55:19,040 NUMBERS MIGHT BE A LITTLE BIT 4070 02:55:19,040 --> 02:55:20,640 OFF BUT YOU CAN SO E THAT MORE 4071 02:55:20,640 --> 02:55:23,320 THAN 50% OF THE DRUGS ARE 4072 02:55:23,320 --> 02:55:26,440 TARGETING MEMBRANE PROTEINS. 4073 02:55:26,440 --> 02:55:28,000 UNFORTUNATELY, RUNNING THE DEL 4074 02:55:28,000 --> 02:55:29,520 SCREEN FOR MEMBRANE PROTEINS IS 4075 02:55:29,520 --> 02:55:32,360 VERY CHALLENGING. SO WHY IS 4076 02:55:32,360 --> 02:55:33,880 THAT? YOU BASICALLY HAVE -- YOU 4077 02:55:33,880 --> 02:55:35,920 HAVE A COUPLE OF CHOICES OF HOW 4078 02:55:35,920 --> 02:55:37,960 YOU RUN THIS SELECTION. IF YOU 4079 02:55:37,960 --> 02:55:39,840 ARE LUCKY AND YOU CAN FORTIFY 4080 02:55:39,840 --> 02:55:40,960 YOUR PROTEIN AND STABLE ENOUGH 4081 02:55:40,960 --> 02:55:44,040 YOU CAN TRY TO DO THAT. HOWEVER, 4082 02:55:44,040 --> 02:55:45,080 UNFORTUNATELY MOST OF THE TIME 4083 02:55:45,080 --> 02:55:47,760 IT IS NOT GOING TO HAPPEN. YOU 4084 02:55:47,760 --> 02:55:49,040 WILL HAVE PA HARD TIME 4085 02:55:49,040 --> 02:55:52,360 FORTIFYING THE PROTEIN, KEEPING 4086 02:55:52,360 --> 02:55:57,320 IT IN SOLUTION AND FOLDED. SO 4087 02:55:57,320 --> 02:55:58,680 ANOTHER OPTION WOULD BE RUNNING 4088 02:55:58,680 --> 02:56:04,080 IT ON USING CELLS. WELL YOU CAN 4089 02:56:04,080 --> 02:56:05,520 MENTION THAT IF YOU WANT TO USE 4090 02:56:05,520 --> 02:56:07,800 CELL FOR DEL SELECTION YOUR 4091 02:56:07,800 --> 02:56:09,080 BACKGROUND WOULD BE MUCH HIGHER 4092 02:56:09,080 --> 02:56:10,440 DUE TO NON-SPECIFIC BINDING BUT 4093 02:56:10,440 --> 02:56:11,160 THIS IS SOMETHING YOU CAN 4094 02:56:11,160 --> 02:56:12,680 ADDRESS WITH THE RIGHT NEGATIVE 4095 02:56:12,680 --> 02:56:14,800 CONTROL, FOR EXAMPLE USING THE 4096 02:56:14,800 --> 02:56:17,040 PARENTAL CELL LINE. HOWEVER, THE 4097 02:56:17,040 --> 02:56:19,560 REAL CHALLENGE HERE COMES FROM 4098 02:56:19,560 --> 02:56:23,040 THE FACT THAT AS I SAID, BALI -- 4099 02:56:23,040 --> 02:56:24,640 BINDING IN DEL IS DRIVEN BY THE 4100 02:56:24,640 --> 02:56:26,920 PROTEIN CONCENTRATION. WHICH 4101 02:56:26,920 --> 02:56:29,600 MEANS THAT CELL BASED SELECTION 4102 02:56:29,600 --> 02:56:31,080 WE WOULD NEED CELLS THAT 4103 02:56:31,080 --> 02:56:34,080 EXPRESSES THE RECEPTOR OF 4104 02:56:34,080 --> 02:56:36,160 INTEREST IN VERY HIGH 4105 02:56:36,160 --> 02:56:38,920 CONCENTRATCONCENTRATION. SO THEA 4106 02:56:38,920 --> 02:56:40,840 COUPLE OF PAPERS OUT THERE I 4107 02:56:40,840 --> 02:56:44,240 WANTED TO MENTION QUICKLY. FOR 4108 02:56:44,240 --> 02:56:46,680 EXAMPLE, DUKE UNIVERSITY IN 4109 02:56:46,680 --> 02:56:48,800 COLLABORATION WITH LEXICON, THEY 4110 02:56:48,800 --> 02:56:53,680 WORKED WITH THE BETA ADRENERGIC 4111 02:56:53,680 --> 02:56:57,120 RECEPTOR, THEY FORTIFY AND ON 4112 02:56:57,120 --> 02:56:59,240 MAGNETE SURFACE AND KEPT THE 4113 02:56:59,240 --> 02:57:01,840 RECEPTOR IN A SOLUTION. ASTRA 4114 02:57:01,840 --> 02:57:04,360 ZENECA USED A SIMILAR METHOD FOR 4115 02:57:04,360 --> 02:57:05,400 THEIR COMPANY BUT IN THIS CASE 4116 02:57:05,400 --> 02:57:08,720 THEY INTRODUCED POINT MUTATIONS 4117 02:57:08,720 --> 02:57:11,920 TO STABILIZE THE PROTEIN IN AN 4118 02:57:11,920 --> 02:57:14,200 ANTIBODY STABLE CONFIRMATION. 4119 02:57:14,200 --> 02:57:17,760 AND AGAIN, THEY USE (INAUDIBLE) 4120 02:57:17,760 --> 02:57:19,520 TO KEEP PROTEIN IN SOLUTION. AS 4121 02:57:19,520 --> 02:57:21,640 I SAID BEFORE THIS IS A POSSIBLE 4122 02:57:21,640 --> 02:57:23,800 ROUTE BUT IT IS VERY LIKELY THAT 4123 02:57:23,800 --> 02:57:26,080 IT IS LIKELY YOU WON'T BE ABLE 4124 02:57:26,080 --> 02:57:27,880 TO EXPRESS AND FORTIFY YOUR 4125 02:57:27,880 --> 02:57:30,880 PROTEIN AND ALSO ALL OF THESE 4126 02:57:30,880 --> 02:57:33,000 DATA CHARTS THAT THE DIFFERENT 4127 02:57:33,000 --> 02:57:34,480 GROUPS USE, THEIR COMPATIBILITY 4128 02:57:34,480 --> 02:57:36,520 WAS NEVER ACTUALLY STUDIED WITH 4129 02:57:36,520 --> 02:57:37,720 DEL SO WE HAVE NO INFORMATION 4130 02:57:37,720 --> 02:57:39,720 ABOUT THEM. THERE ARE A COUPLE 4131 02:57:39,720 --> 02:57:42,960 OF PUBLICATIONS THAT USE SLIDE 4132 02:57:42,960 --> 02:57:45,960 CELLS AS WELL. SO FOR EXAMPLE 4133 02:57:45,960 --> 02:57:49,720 HERE, THE GROUP EXPRESSED FOLATE 4134 02:57:49,720 --> 02:57:51,880 RECEPTOR IN CELLS AND FOLATE HAD 4135 02:57:51,880 --> 02:57:54,320 A DNA TAG ATTACHED TO IT. SO 4136 02:57:54,320 --> 02:57:57,240 THIS DNA TAG ESSENTIALLY 4137 02:57:57,240 --> 02:57:59,200 HYBRIDIZES WITH THE DEL TAG 4138 02:57:59,200 --> 02:58:01,200 ITSELF AND ESSENTIALLY GUIDING 4139 02:58:01,200 --> 02:58:03,440 THE COMPOUNDS TO THE RECEPTOR 4140 02:58:03,440 --> 02:58:05,960 AND INITIATING THE BINDING. SO 4141 02:58:05,960 --> 02:58:07,280 IN THIS CASE THEY DIDN'T NEED TO 4142 02:58:07,280 --> 02:58:08,640 EXPRESS THE PROTEIN AT HIGH 4143 02:58:08,640 --> 02:58:12,160 CONCENTRATION. AND THERE WAS A 4144 02:58:12,160 --> 02:58:15,600 NICE ELEGANT WORK FROM GSK 4145 02:58:15,600 --> 02:58:19,280 WORKING WITH NK 3 RECEPTOR, 4146 02:58:19,280 --> 02:58:23,600 WHERE THEY TRANSDUCED HEP 293 4147 02:58:23,600 --> 02:58:24,760 CELL WITHBACTER VIRUS AND THEY 4148 02:58:24,760 --> 02:58:27,960 WERE ABLE TO GET 500 RECEPTORS 4149 02:58:27,960 --> 02:58:29,520 PER CELL, VERY HIGH 4150 02:58:29,520 --> 02:58:31,040 CONCENTRATION AND THEN RAN THE 4151 02:58:31,040 --> 02:58:33,840 DEL SELECTION ON THESE SLIVE 4152 02:58:33,840 --> 02:58:37,160 CELLS. THE GSK PAPER SUGGESTS TO 4153 02:58:37,160 --> 02:58:39,840 HAVE AT LEAST 100,000 COPIES PER 4154 02:58:39,840 --> 02:58:41,520 CELL, BUT THE MORE YOU HAVE THE 4155 02:58:41,520 --> 02:58:43,000 BETTER CHANCE YOU ARE GOING TO 4156 02:58:43,000 --> 02:58:45,680 GET FOR DEL SELECTION. AND THIS 4157 02:58:45,680 --> 02:58:47,280 METHOD IS ACTUALLY SOMETHING 4158 02:58:47,280 --> 02:58:48,920 THAT NOW WE ARE EXPLORING AT 4159 02:58:48,920 --> 02:58:50,400 PFIZER AS WELL AND TRYING TO 4160 02:58:50,400 --> 02:58:55,360 LEARN MORE ABOUT IT. NOW, JUST A 4161 02:58:55,360 --> 02:58:58,560 QUICK SUMMARY. DEL SCREENS ARE 4162 02:58:58,560 --> 02:58:59,720 RELATIVELY LOW COST ONCE YOU 4163 02:58:59,720 --> 02:59:02,440 HAVE ESTABLISH YOUR LIBRARY AND 4164 02:59:02,440 --> 02:59:03,760 YOUR PLATFORM AND CAN BE 4165 02:59:03,760 --> 02:59:05,000 COMPLETED IN JUST A COUPLE OF 4166 02:59:05,000 --> 02:59:07,000 MONTHS AS WELL. IT IS USUALLY 4167 02:59:07,000 --> 02:59:09,880 RESULTS IN MINIMAL OFF DNA 4168 02:59:09,880 --> 02:59:12,120 SYNTHESIS SO TEN COMPOUNDS OR 4169 02:59:12,120 --> 02:59:14,320 SO. MAKE SURE THAT YOU VALIDATE 4170 02:59:14,320 --> 02:59:16,240 YOUR REAGENTS BEFORE DEL 4171 02:59:16,240 --> 02:59:18,240 SCREENING, THAT SHOULD BE THE 4172 02:59:18,240 --> 02:59:21,200 CORNERSTONE OF YOUR WORK FLOW. 4173 02:59:21,200 --> 02:59:24,920 AND USING MPO SCORING AND ON DNA 4174 02:59:24,920 --> 02:59:27,240 RESYNTHESIS YOU CAN INCREASE 4175 02:59:27,240 --> 02:59:28,120 CONFIRMATION THE HIT 4176 02:59:28,120 --> 02:59:29,760 CONFIRMATION RATE AND ENSURE YOU 4177 02:59:29,760 --> 02:59:31,120 HAVE A GOOD DRUG LIKE PROPERTY 4178 02:59:31,120 --> 02:59:35,360 AT THE END OF IT. NOW, DEL IS 4179 02:59:35,360 --> 02:59:39,000 REALLY A TEAM WORK AND I WANT TO 4180 02:59:39,000 --> 02:59:40,600 ACKNOWLEDGE EVERYBODY AT PFIZER 4181 02:59:40,600 --> 02:59:44,720 AND ALSO COLLABORATORS AT HIT 4182 02:59:44,720 --> 02:59:47,720 GEN. AS I SAID DURING THE DEL 4183 02:59:47,720 --> 02:59:50,840 SELECTION BIOLOGICAL CHEMISTS 4184 02:59:50,840 --> 02:59:54,480 HAVE TO WORK TOGETHER TO ENSURE 4185 02:59:54,480 --> 02:59:55,320 SUCCESSFUL COMPANY. THANK YOU 4186 02:59:55,320 --> 02:59:57,120 FOR YOUR ATTENTION AND HAPPY TO 4187 02:59:57,120 --> 02:59:57,560 ANSWER ANY QUESTIONS. 4188 02:59:57,560 --> 03:00:02,320 [APPLAUSE] 4189 03:00:02,320 --> 03:00:04,840 >>THANK YOU SO MUCH SOFIA FOR A 4190 03:00:04,840 --> 03:00:08,720 WONDERFUL TALK. QUESTIONS FOR 4191 03:00:08,720 --> 03:00:12,520 SOPHIA? 4192 03:00:12,520 --> 03:00:14,000 >>DO YOU MIND APPROACHING THE 4193 03:00:14,000 --> 03:00:17,640 MIC OVER THERE? 4194 03:00:17,640 --> 03:00:18,960 (OFF MIC) 4195 03:00:18,960 --> 03:00:20,240 >>THAT WAS REALLY EXCELLENT 4196 03:00:20,240 --> 03:00:22,640 TALK AND EXPLAINED A LOT ABOUT 4197 03:00:22,640 --> 03:00:24,680 DEL LIBRARIES WHICH ARE REALLY 4198 03:00:24,680 --> 03:00:26,080 INTERESTING. ONE THING THAT I 4199 03:00:26,080 --> 03:00:29,560 HAVE NEVER QUITE UNDERSTOOD IS 4200 03:00:29,560 --> 03:00:30,960 HOW AFTER AMPLIFICATION AFTER 4201 03:00:30,960 --> 03:00:34,840 THE FIRST ROUND YOU RESYNTHESIZE 4202 03:00:34,840 --> 03:00:36,520 YOUR COMPOUND LIBRARY ALONG WITH 4203 03:00:36,520 --> 03:00:39,160 THE PCR PRODUCT. CAN YOU DISCUSS 4204 03:00:39,160 --> 03:00:41,800 THAT? 4205 03:00:41,800 --> 03:00:43,400 >>I'M NOT SURE I UNDERSTAND 4206 03:00:43,400 --> 03:00:46,560 YOUR QUESTION. SO AFTER YOU DID 4207 03:00:46,560 --> 03:00:49,240 YOUR ROUND OF DEL SELECTIONS? 4208 03:00:49,240 --> 03:00:49,800 >>YES. 4209 03:00:49,800 --> 03:00:51,440 >>OKAY. SO AFTER THAT YOU 4210 03:00:51,440 --> 03:00:53,480 BASICALLY AMPLIFY YOUR DNA TAGS 4211 03:00:53,480 --> 03:00:55,520 DURING PCR. AND THEN YOU HAVE TO 4212 03:00:55,520 --> 03:00:58,400 TAKE IT INTO NEXT GENERATION 4213 03:00:58,400 --> 03:00:59,880 SEQUENCING. SO THERE WERE A 4214 03:00:59,880 --> 03:01:01,520 COUPLE OF STEPS INVOLVED HERE SO 4215 03:01:01,520 --> 03:01:04,200 AFTER PCR AMPLIFICATION YOU HAVE 4216 03:01:04,200 --> 03:01:06,120 TO ADD SOME KIND OF ADAPTER THAT 4217 03:01:06,120 --> 03:01:08,120 WILL ALLOW YOUR DNA TAG TO 4218 03:01:08,120 --> 03:01:09,840 ATTACH TO THE SEQUENCING CHIP AS 4219 03:01:09,840 --> 03:01:12,840 WELL. BUT AT THAT -- AT THIS 4220 03:01:12,840 --> 03:01:15,200 POINT YOU ARE LOOKING AT THIS 4221 03:01:15,200 --> 03:01:17,560 LIKE DNA SEQUENCE. YOU KIND OF 4222 03:01:17,560 --> 03:01:18,680 -- YOU FORGET ABOUT THIS BEING 4223 03:01:18,680 --> 03:01:20,400 COMPOUNDS, YOU ARE GOING TO JUST 4224 03:01:20,400 --> 03:01:22,960 AMPLIFY YOUR SEQUENCE USING PCR 4225 03:01:22,960 --> 03:01:26,360 AND WORK WITH THAT LATER ON. 4226 03:01:26,360 --> 03:01:30,560 >>THEN SO THE SECOND ROUND OF 4227 03:01:30,560 --> 03:01:33,280 SELECTION, YOU REPEAT THE 96 4228 03:01:33,280 --> 03:01:34,080 WELL PLATE COMBINATORIAL 4229 03:01:34,080 --> 03:01:34,360 CHEMISTRY? 4230 03:01:34,360 --> 03:01:37,520 >>NO. SO IN THE SECOND -- AFTER 4231 03:01:37,520 --> 03:01:40,160 THE FIRST ROUND OF SELECTION, 4232 03:01:40,160 --> 03:01:42,520 YOU ADD TO YOUR COMPOUNDS. YOU 4233 03:01:42,520 --> 03:01:44,120 HAVE SOME YOU ADD THEN YOU TAKE 4234 03:01:44,120 --> 03:01:45,920 THE EXACT SAME COMPOUNDS AND 4235 03:01:45,920 --> 03:01:48,040 TAKE INTO YOUR SECOND ROUND OF 4236 03:01:48,040 --> 03:01:48,880 SELECTION AS AN INPUT. 4237 03:01:48,880 --> 03:01:50,440 >>OKAY. THANKS. 4238 03:01:50,440 --> 03:01:53,920 >>YEP. 4239 03:01:53,920 --> 03:01:58,400 >>THANK YOU FOR YOUR TALK. SO I 4240 03:01:58,400 --> 03:01:59,640 WANT TO KNOW MORE ABOUT BUILD 4241 03:01:59,640 --> 03:02:01,520 SCREENING FOR THE DNA BINDING 4242 03:02:01,520 --> 03:02:03,960 PROTEINS. SO SOME OF THE 4243 03:02:03,960 --> 03:02:06,800 PROTEINS COULD HAVE DNA BINDING 4244 03:02:06,800 --> 03:02:08,720 POCKET OR WHATEVER YOU CALL, SO 4245 03:02:08,720 --> 03:02:10,080 DO YOU THINK THE CHANCE OF 4246 03:02:10,080 --> 03:02:13,400 GETTING MORE OFF TARGET OR I 4247 03:02:13,400 --> 03:02:14,040 WANT TO KNOW MORE ABOUT THAT. 4248 03:02:14,040 --> 03:02:14,520 THANK YOU. 4249 03:02:14,520 --> 03:02:16,320 >>THAT IS ACTUALLY REALLY GOOD 4250 03:02:16,320 --> 03:02:18,160 QUESTION. WE HAD A COUPLE OF 4251 03:02:18,160 --> 03:02:20,120 TARGETS WHICH WERE EITHER DNA OR 4252 03:02:20,120 --> 03:02:22,880 RNA BINDING PROTEIN. IN THAT 4253 03:02:22,880 --> 03:02:25,640 CASE WE WERE LUCKY, SO WE DID 4254 03:02:25,640 --> 03:02:28,840 SEE BINDING TO DNA TAG BUT THAT 4255 03:02:28,840 --> 03:02:30,880 BINDING WAS NON-SPECIFIC. SO 4256 03:02:30,880 --> 03:02:33,640 THAT MEANS IT WASN'T ENRICHED 4257 03:02:33,640 --> 03:02:34,880 DURING SELECTION AND IT IS JUST 4258 03:02:34,880 --> 03:02:36,560 RESULTING IN HIGH BACKGROUND 4259 03:02:36,560 --> 03:02:38,120 WHICH WE COULD GET RID OF DURING 4260 03:02:38,120 --> 03:02:41,440 THE DATA ANALYSIS. HOWEVER, FOR 4261 03:02:41,440 --> 03:02:44,200 EXAMPLE, HITGEN OUR 4262 03:02:44,200 --> 03:02:45,440 COLLABORATOR, THEY HAVEN'T 4263 03:02:45,440 --> 03:02:48,200 ESTABLISHED WORK FLOW, WELL THEY 4264 03:02:48,200 --> 03:02:50,360 ACTUALLY ADD -- SO IN TERMS OF 4265 03:02:50,360 --> 03:02:51,920 RNA BINDING PROTEINS YOUR 4266 03:02:51,920 --> 03:02:54,320 PROBLEM IS THAT THE DNA TAG HAS 4267 03:02:54,320 --> 03:02:57,320 A SINGLE CHAIN REGION AS WELL. 4268 03:02:57,320 --> 03:02:59,880 AND HITGEN CAME UP WITH AN 4269 03:02:59,880 --> 03:03:02,040 APPROACH WHERE THEY HYDROLYZE 4270 03:03:02,040 --> 03:03:05,400 THE SINGLE CHAIN INTO SMALL RNA 4271 03:03:05,400 --> 03:03:07,640 PIECES AND THEN RUN THE DEL 4272 03:03:07,640 --> 03:03:08,640 SELECTION ON THAT, SO THEY WERE 4273 03:03:08,640 --> 03:03:10,320 ABLE TO REDUCE THE BACKGROUND. 4274 03:03:10,320 --> 03:03:12,920 BUT AS I SAID IN OUR CASE, WE 4275 03:03:12,920 --> 03:03:14,280 NEVER -- WE DIDN'T HAVE PROBLEM 4276 03:03:14,280 --> 03:03:16,360 BECAUSE BINDING WAS ALWAYS 4277 03:03:16,360 --> 03:03:18,360 NON-SPECIFIC SO YES IT INCREASED 4278 03:03:18,360 --> 03:03:19,720 OUR BACKGROUND BUT WE COULD GET 4279 03:03:19,720 --> 03:03:20,720 RID OF THAT DURING DATA 4280 03:03:20,720 --> 03:03:24,400 ANALYSIS. 4281 03:03:24,400 --> 03:03:26,560 >>ACTUALLY HAVE TWO QUESTIONS. 4282 03:03:26,560 --> 03:03:37,760 SO THE FIRST ONE, SO (INAU -- 4283 03:03:40,960 --> 03:03:44,280 >>GOT YOU. SORRY ABOUT THAT. I 4284 03:03:44,280 --> 03:03:45,560 HAVE TWO QUESTIONS. FIRST WHEN 4285 03:03:45,560 --> 03:03:47,040 YOU POOL YOUR LIBRARIES TOGETHER 4286 03:03:47,040 --> 03:03:49,800 LIKE YOU WORRY ABOUT THE SIZE 4287 03:03:49,800 --> 03:03:51,840 DIFFERENCE OF THE LIBRARIES? SAY 4288 03:03:51,840 --> 03:03:54,960 LIKE YOU POOL A LIBRARY THAT HAS 4289 03:03:54,960 --> 03:03:56,760 BEADS OF MOLECULES AND YOU PUT 4290 03:03:56,760 --> 03:03:58,320 IT TOGETHER WITH A LIBRARY THAT 4291 03:03:58,320 --> 03:03:59,440 ONLY HAS MILLIONS OF MOLECULE, 4292 03:03:59,440 --> 03:04:01,360 DO YOU WORRY THAT THE LARGE 4293 03:04:01,360 --> 03:04:02,240 LIBRARY WILL BE DILUTED IN THIS 4294 03:04:02,240 --> 03:04:02,960 WAY? 4295 03:04:02,960 --> 03:04:05,400 >>THAT IS A GOOD POINT. AND 4296 03:04:05,400 --> 03:04:07,720 THEN WHEN YOU POOL THEM TOGETHER 4297 03:04:07,720 --> 03:04:08,960 FOR SELECTION, WE PULL IT IN A 4298 03:04:08,960 --> 03:04:13,040 WAY THAT AT THE END EVERY SINGLE 4299 03:04:13,040 --> 03:04:14,640 COMPOUND WE HAVE COPY, SO THAT 4300 03:04:14,640 --> 03:04:15,560 IS NOT SOMETHING YOU HAVE TO 4301 03:04:15,560 --> 03:04:16,920 WORRY ABOUT BECAUSE THAT IS A 4302 03:04:16,920 --> 03:04:17,280 GOOD POINT. 4303 03:04:17,280 --> 03:04:19,720 >>MY SECOND QUESTION IS SO FOR 4304 03:04:19,720 --> 03:04:23,720 THE CELL BASED DEL ASSAY, SO CAN 4305 03:04:23,720 --> 03:04:26,520 YOU ADD SOME COMMENTS ON LIKE 4306 03:04:26,520 --> 03:04:27,760 POSSIBLE NO TARGET CONTROL LIKE 4307 03:04:27,760 --> 03:04:29,120 YOU JUST USE THE SAME CELL LINE 4308 03:04:29,120 --> 03:04:30,560 THAT DOESN'T EXPRESS THE PROTEIN 4309 03:04:30,560 --> 03:04:34,280 OR YOU HAVE LIKE OTHER -- LIKE 4310 03:04:34,280 --> 03:04:35,480 NO TARGET CONTROLS? 4311 03:04:35,480 --> 03:04:37,640 >>YEAH, SO AS I SAID WE ARE 4312 03:04:37,640 --> 03:04:38,880 JUST GETTING TO THIS SPACE AND 4313 03:04:38,880 --> 03:04:40,800 WE ARE LEARNING ABOUT IT. BUT 4314 03:04:40,800 --> 03:04:43,000 RIGHT NOW OUR STRATEGY IS TO USE 4315 03:04:43,000 --> 03:04:45,120 THE PARENTAL CELL LINE OR THE 4316 03:04:45,120 --> 03:04:47,120 CELL LINE THAT WAS TRANSDUCED 4317 03:04:47,120 --> 03:04:48,440 NEGATIVE CONTROL VIRUS OR 4318 03:04:48,440 --> 03:04:49,800 SOMETHING LIKE THAT. 4319 03:04:49,800 --> 03:04:54,480 >>GOT YOU. THANKS. 4320 03:04:54,480 --> 03:04:58,480 >>SO GPCR PROJECTS THAT 4321 03:04:58,480 --> 03:05:00,280 INTERESTING DO THEY TALK ABOUT 4322 03:05:00,280 --> 03:05:02,240 WHETHER THEY HAVE TO CALL 4323 03:05:02,240 --> 03:05:04,360 AMPLIFY OR CO-TRANSFECT IN THE G 4324 03:05:04,360 --> 03:05:05,880 ALPHA COMPONENT WITH THE ACTUAL 4325 03:05:05,880 --> 03:05:06,520 RECEPTOR ITSELF? 4326 03:05:06,520 --> 03:05:09,480 >>YES, SO IN THESE PAPERS, THEY 4327 03:05:09,480 --> 03:05:11,640 DIDN'T DO THAT. BUT THAT IS 4328 03:05:11,640 --> 03:05:13,000 ACTUALLY AN OPTION AS WELL THAT 4329 03:05:13,000 --> 03:05:15,000 WE ARE EXPLORING. SO THAT IS A 4330 03:05:15,000 --> 03:05:19,520 VERY GOOD POINT. YES. 4331 03:05:19,520 --> 03:05:21,440 >>ALL RIGHT. THANK YOU VERY 4332 03:05:21,440 --> 03:05:28,880 MUCH, SOPHIA, AGAIN. SO COUPLE 4333 03:05:28,880 --> 03:05:30,520 OF QUICK ANNOUNCEMENTS BEFORE WE 4334 03:05:30,520 --> 03:05:33,320 GO TO LUNCH, ONE THING I WANT TO 4335 03:05:33,320 --> 03:05:35,640 REMIND YOU ALL THAT WE ACTUALLY 4336 03:05:35,640 --> 03:05:38,160 HAVE TOPIC FOCUSED WORKSHOP ON 4337 03:05:38,160 --> 03:05:41,880 DEL TWO DAYS OF VIDEOS YOU CAN 4338 03:05:41,880 --> 03:05:47,200 WATCH ONLINE, ON DEMAND NIH 4339 03:05:47,200 --> 03:05:49,760 VIDEOCAST WEBSITE, WE'LL SHARE 4340 03:05:49,760 --> 03:05:50,840 ALL THESE IF THAT IS SOMETHING 4341 03:05:50,840 --> 03:05:52,520 OF INTEREST AND YOU WANT TO DIVE 4342 03:05:52,520 --> 03:05:56,240 MORE INTO DNA LIBRARIES. SO WE 4343 03:05:56,240 --> 03:05:57,960 ARE -- WE ARE GOING TO TAKE AN 4344 03:05:57,960 --> 03:06:00,520 HOUR BREAK NOW. WE WILL 4345 03:06:00,520 --> 03:06:02,320 RECONVENE AT 1:30. WE HAVE 4346 03:06:02,320 --> 03:06:03,920 EXCELLENT TALKS THIS AFTERNOON. 4347 03:06:03,920 --> 03:06:07,680 SO COME BACK, WE WILL HAVE TALKS 4348 03:06:07,680 --> 03:06:09,840 ON HTS DATA ANALYSIS. DRUG 4349 03:06:09,840 --> 03:06:11,920 SCREENING USING 3D TISSUE 4350 03:06:11,920 --> 03:06:14,000 MODELS. COUPLE OF MORE EXCITING 4351 03:06:14,000 --> 03:06:14,680 TALKS AWAITING YOU THIS 4352 03:06:14,680 --> 03:06:17,600 AFTERNOON. WE WILL SEE YOU AT 4353 03:06:17,600 --> 03:06:19,160 1:30. THANK YOU. 4354 03:06:19,160 --> 03:06:21,720 WELCOME BACK, 4355 03:06:21,720 --> 03:06:24,240 EVERYONE. OUR NEXT SPEAKER IS 4356 03:06:24,240 --> 03:06:29,760 DR. THOMAS CHUNG. DR. CHUNG IS 4357 03:06:29,760 --> 03:06:31,680 THE DIRECTOR OF TRANSLATIONAL 4358 03:06:31,680 --> 03:06:34,840 PROGRAMS OUTREACH AT THE PREBYS 4359 03:06:34,840 --> 03:06:39,040 CENTER FOR CHEMICAL GENOMICS AT 4360 03:06:39,040 --> 03:06:40,960 SANFORD PREBYS MEDICAL DISCOVERY 4361 03:06:40,960 --> 03:06:44,200 INSTITUTE SVP IN LA JOLLA, 4362 03:06:44,200 --> 03:06:45,880 CALIFORNIA. DR. CHUNG IS A LONG 4363 03:06:45,880 --> 03:06:49,080 STANDING EDITORIAL BOARD MEMBER 4364 03:06:49,080 --> 03:06:52,240 OF THE AGM AND HAS MULTIPLE 4365 03:06:52,240 --> 03:06:54,360 RECOGNIZED CHAPTERS IN THE AGM 4366 03:06:54,360 --> 03:06:55,920 SO WE ARE EXCITED TO HAVE HIM 4367 03:06:55,920 --> 03:06:57,960 HERE TODAY. HIS TALK IS 4368 03:06:57,960 --> 03:07:00,600 ENTITLED BASIC ASSAYS 4369 03:07:00,600 --> 03:07:02,080 STATISTICS, DATA ANALYSIS AND 4370 03:07:02,080 --> 03:07:04,160 RULES OF THUMB. TC, THE FLOOR IS 4371 03:07:04,160 --> 03:07:10,120 YOURS. 4372 03:07:10,120 --> 03:07:14,280 [APPLAUSE] 4373 03:07:14,280 --> 03:07:17,080 >>I HAVE HAD A LONG HISTORY 4374 03:07:17,080 --> 03:07:19,440 WITH THE SOAS AND THE AGM SO IT 4375 03:07:19,440 --> 03:07:23,320 IS A PLEASURE. TODAY I'M TRYING 4376 03:07:23,320 --> 03:07:28,720 TO TALK BASIC FOUNDATIONS, 4377 03:07:28,720 --> 03:07:30,760 ADEQUATELY COVERED DEVELOPMENT 4378 03:07:30,760 --> 03:07:33,200 AND VARIOUS THINGS. WHAT I'M 4379 03:07:33,200 --> 03:07:34,440 HERE TO GIVE YOU IS RULES OF 4380 03:07:34,440 --> 03:07:36,400 THUMB, WHAT I'M PRESENTING IS 4381 03:07:36,400 --> 03:07:38,400 REALLY A DILUTED STATISTICS 4382 03:07:38,400 --> 03:07:42,520 CLASS, IT IS REALLY A REAL 4383 03:07:42,520 --> 03:07:44,080 STATISTICIAN PROBABLY CRUCIFY 4384 03:07:44,080 --> 03:07:47,240 ME. REALLY I WANT TO HELP 4385 03:07:47,240 --> 03:07:48,560 EVERYONE THINK HOW WHAT YOU MEAN 4386 03:07:48,560 --> 03:07:52,760 BY ASSAY ROBUSTNESS. AND WHAT 4387 03:07:52,760 --> 03:07:54,560 VALIDATION MEANS IN TERMS OF 4388 03:07:54,560 --> 03:07:57,600 ROBUSTNESS OF ASSAY. SO YOU HAVE 4389 03:07:57,600 --> 03:07:58,920 HEARD SEVERAL TIMES THERE'S 4390 03:07:58,920 --> 03:08:01,720 ASSAYS AND THE ASSAYS ARE 4391 03:08:01,720 --> 03:08:04,240 EMPLOYED IN HTS HEALTHCARE YOU 4392 03:08:04,240 --> 03:08:05,320 HAVE ONE CHANCE TO TEST THAT 4393 03:08:05,320 --> 03:08:07,680 COMPOUND SO YOU WANT TO BE AT 4394 03:08:07,680 --> 03:08:10,400 LEAST SURE THAT THE ASSAY ITSELF 4395 03:08:10,400 --> 03:08:15,280 WASN'T -- WAS BEEHIVING W BEHAVO 4396 03:08:15,280 --> 03:08:18,240 THEY HAVE TO BE RIGOROUSLY 4397 03:08:18,240 --> 03:08:20,200 VALIDATED THESE ASSAYS FOR 4398 03:08:20,200 --> 03:08:21,520 VIABILITY AS WELL AS 4399 03:08:21,520 --> 03:08:23,240 PHARMACOLOGICAL EVENTS NOT JUST 4400 03:08:23,240 --> 03:08:24,480 IN THE DISEASE STATE BUT 4401 03:08:24,480 --> 03:08:28,720 ACTUALLY IN TERMS OF THE 4402 03:08:28,720 --> 03:08:30,240 PHENOMENOLOGICAL AFFECT WHEN 4403 03:08:30,240 --> 03:08:34,920 COMPOUND RICH REACHES THE TARGEU 4404 03:08:34,920 --> 03:08:36,800 HEARD FROM NATHAN AND TIM ABOUT 4405 03:08:36,800 --> 03:08:38,960 VERIFYING REAGENTS STABILITY FOR 4406 03:08:38,960 --> 03:08:41,640 STORAGE AS WELL AS DURING THE 4407 03:08:41,640 --> 03:08:45,520 ASSAY. AND THINGS LIKE ROBUST 4408 03:08:45,520 --> 03:08:48,600 FREEZE THAWING AND -- TRY THIS. 4409 03:08:48,600 --> 03:08:52,600 THAT'S GOOD. FREEZE THAWING AND 4410 03:08:52,600 --> 03:08:54,720 LONG TERM TEMPERATURES. WITH 4411 03:08:54,720 --> 03:08:56,200 THESE ASSAYS WHEN YOU START 4412 03:08:56,200 --> 03:08:57,680 DOING BIG CAMPAIGNS, YOU ARE 4413 03:08:57,680 --> 03:09:00,280 TRYING TO DO IT IN RAPID 4414 03:09:00,280 --> 03:09:02,200 SUCCESSION AND YOU HAVE TO WORK 4415 03:09:02,200 --> 03:09:05,760 WITH -- YOU HAVE TO HAVE THE 4416 03:09:05,760 --> 03:09:06,360 STOCK SOLUTION MADE TEMPORARY 4417 03:09:06,360 --> 03:09:07,760 WORKING STOCKS AND U YOU WANT TO 4418 03:09:07,760 --> 03:09:09,240 MAKE SURE THEY ARE USEFUL AND 4419 03:09:09,240 --> 03:09:12,240 STABLE FOR THAT. WHAT DO YOU DO 4420 03:09:12,240 --> 03:09:13,480 WITH THE LEFT OVERS? SOMETIMES 4421 03:09:13,480 --> 03:09:15,480 YOU JUST THROW THEM AWAY BUT 4422 03:09:15,480 --> 03:09:16,880 OCCASIONALLY YOU CAN REFREEZE 4423 03:09:16,880 --> 03:09:18,000 THEM AND FIND THAT YOU CAN USE 4424 03:09:18,000 --> 03:09:21,120 IT FOR FOLLOW UP STUDIES. FOR 4425 03:09:21,120 --> 03:09:23,000 ASSAY ROBUSTNESS AND PERFORMANCE 4426 03:09:23,000 --> 03:09:24,840 OTHER THINGS TO DO IS YOU WANT 4427 03:09:24,840 --> 03:09:26,480 TO IDENTIFY AND ELIMINATE THE 4428 03:09:26,480 --> 03:09:29,120 SOURCES OF SYSTEMIC ERRORS SO 4429 03:09:29,120 --> 03:09:30,400 YOU HAVE HEARD ABOUT CHECKING 4430 03:09:30,400 --> 03:09:34,360 FOR DMS COMPATIBILITY. THE OTHER 4431 03:09:34,360 --> 03:09:36,400 IS TO CHECK PHYSICAL UNIFORMITY, 4432 03:09:36,400 --> 03:09:37,760 NOW DAYS THE VENDORS ARE GOOD 4433 03:09:37,760 --> 03:09:39,440 ABOUT THOSE. IN THE OLD DAYS 4434 03:09:39,440 --> 03:09:41,080 PLATES WARPED DEPENDING THE 4435 03:09:41,080 --> 03:09:42,800 VENDOR YOU WENT TO. BUT THERE IS 4436 03:09:42,800 --> 03:09:46,240 OTHER EFFECTS LIKE IF YOU THINGS 4437 03:09:46,240 --> 03:09:48,800 EDGES OF PLACE TEND TO BE HEATED 4438 03:09:48,800 --> 03:09:50,480 SOONER THAN THE CENTERS OF THE 4439 03:09:50,480 --> 03:09:53,240 PLATE BECAUSE PLASTIC IS VERY 4440 03:09:53,240 --> 03:09:55,880 PORE CONDUCTOR OF HEAT. 4441 03:09:55,880 --> 03:09:58,000 DEPENDING UPON YOUR LIQUID 4442 03:09:58,000 --> 03:10:00,400 HANDLING SYSTEMS YOU CAN SEE 4443 03:10:00,400 --> 03:10:02,200 SYSTEMIC CLOG TIPS WILL LEAF A 4444 03:10:02,200 --> 03:10:03,480 STRIPING. I WILL SHOW YOU 4445 03:10:03,480 --> 03:10:05,120 EXAMPLES LATER. YOU WANT TO 4446 03:10:05,120 --> 03:10:07,080 CHECK FOR INNER PLATE AND 4447 03:10:07,080 --> 03:10:09,760 INTERDAY VARIATIONS, TRYING TO 4448 03:10:09,760 --> 03:10:12,560 MINIMIZE THAT. THERE'S WAYS OF 4449 03:10:12,560 --> 03:10:13,720 USING DIFFERENT TYPES OF 4450 03:10:13,720 --> 03:10:16,480 STATISTICS FOR THAT. THE OTHER 4451 03:10:16,480 --> 03:10:17,840 THING IS YOU WANT TO REALLY 4452 03:10:17,840 --> 03:10:19,480 CHECK THAT YOU ARE ON THE LINEAR 4453 03:10:19,480 --> 03:10:22,040 RANGE IF ENZYMATIC OR BINDING 4454 03:10:22,040 --> 03:10:25,600 ASSAY YOU ARE LINEAR PART OF THE 4455 03:10:25,600 --> 03:10:26,480 RESPONSE CURVE WHERE IT IS THE 4456 03:10:26,480 --> 03:10:27,760 MOST SENSITIVE RATHER THAN 4457 03:10:27,760 --> 03:10:34,800 PLATEAUING WHERE RECENTLY 4458 03:10:34,800 --> 03:10:36,200 ASIMILAR TAUPE. SO YOU ARE 4459 03:10:36,200 --> 03:10:37,880 TRYING TO DEAL WITH RANDOM 4460 03:10:37,880 --> 03:10:39,760 ERRORS AN ELIMINATING THE 4461 03:10:39,760 --> 03:10:40,480 SOURCES THINGS YOU CAN CORRECT 4462 03:10:40,480 --> 03:10:42,680 FOR. THE REASON FOR THAT IS 4463 03:10:42,680 --> 03:10:44,600 RANDOM ERRORS YOU CAN'T DO 4464 03:10:44,600 --> 03:10:46,280 ANYTHING ABOUT BUT YOU CAN TREAT 4465 03:10:46,280 --> 03:10:52,200 THEM STATISTICALLY. SO TYPICAL 4466 03:10:52,200 --> 03:10:54,400 ASSAY IS YO YOU HAVE A PRIMARY 4467 03:10:54,400 --> 03:10:56,920 ASSAY THAT MAPS BASICALLY LARGE 4468 03:10:56,920 --> 03:10:58,400 NUMBERS OF COMPOUNDS TO 4469 03:10:58,400 --> 03:11:00,280 BIOLOGICAL ASSAY, YOU HAVE TO 4470 03:11:00,280 --> 03:11:01,960 DEFINE HITS, THRESHOLD. AND 4471 03:11:01,960 --> 03:11:03,640 FOLLOWED BY HITS YOU WILL 4472 03:11:03,640 --> 03:11:05,240 SECONDARY ASSAYS THESE 4473 03:11:05,240 --> 03:11:07,120 ORTHOGONAL ASSAYS, THE HIT 4474 03:11:07,120 --> 03:11:09,960 TRIAGE THAT WAS DISCUSSED FROM 4475 03:11:09,960 --> 03:11:12,080 SEVERAL SPEAKERS. THIS IS A VERY 4476 03:11:12,080 --> 03:11:14,840 TYPICAL FORMAT IF YOU ARE 96 4477 03:11:14,840 --> 03:11:17,280 FORMAT YOU HAVE TYPICALLY MOST 4478 03:11:17,280 --> 03:11:19,680 PEOPLE USE A FIRST TWO COLUMNS 4479 03:11:19,680 --> 03:11:23,080 AS EMPTY OR FIRST AND LAST 4480 03:11:23,080 --> 03:11:25,120 COLUMN IS EMPTY. AND THESE ARE 4481 03:11:25,120 --> 03:11:26,680 WHERE YOUR CONTROLS ARE PUT, AND 4482 03:11:26,680 --> 03:11:28,800 WHEN YOUR CONTROLS ARE PUT, THEY 4483 03:11:28,800 --> 03:11:30,240 GIVE YOU THE HIGH SIGNAL AND THE 4484 03:11:30,240 --> 03:11:33,800 LOW SIGNAL. AND IN THE PAST 4485 03:11:33,800 --> 03:11:35,400 VARIOUS ASSAY PARAMETERS HAVE 4486 03:11:35,400 --> 03:11:38,600 BEEN CALCULATED FOR THESE SUCH 4487 03:11:38,600 --> 03:11:40,720 AS NORMALIZE INHIBITION, THAT'S 4488 03:11:40,720 --> 03:11:42,480 WHAT WE DO, WE ACTUALLY 4489 03:11:42,480 --> 03:11:45,920 NORMALIZE IT TO HIGH AND LOW, 4490 03:11:45,920 --> 03:11:47,680 TAKE IT AS PERCENTAGE OF THAT, 4491 03:11:47,680 --> 03:11:49,760 PUT IT ON THE SAME SCALE TO 4492 03:11:49,760 --> 03:11:51,920 COMPARE THINGS. WE USE A Z SCORE 4493 03:11:51,920 --> 03:11:53,360 WHICH IS FORMALLY UNITS OF 4494 03:11:53,360 --> 03:11:54,640 STANDARD DEVIATION AWAY FROM THE 4495 03:11:54,640 --> 03:11:57,960 MEAN, THAT'S SOMETHING IS, AND 4496 03:11:57,960 --> 03:12:00,200 YOU USE THAT IN TERMS -- YOU 4497 03:12:00,200 --> 03:12:01,680 OFTEN USE THAT WHEN YOU DON'T 4498 03:12:01,680 --> 03:12:05,440 HAVE A HIGH AND LOW CONTROL 4499 03:12:05,440 --> 03:12:07,400 ABILITY CONTROL AND MORE 4500 03:12:07,400 --> 03:12:08,200 COMPLICATED THINGS CAN BE SCORE. 4501 03:12:08,200 --> 03:12:10,080 SO THESE ARIST H I CAN ARE. 4502 03:12:10,080 --> 03:12:12,080 THESE EXIST BECAUSE BUT MOST 4503 03:12:12,080 --> 03:12:14,080 DONE TALK ABOUT THE B SCORE. YOU 4504 03:12:14,080 --> 03:12:18,680 DO NPIs AND PEOPLE OFTEN TALK 4505 03:12:18,680 --> 03:12:21,200 Z SCORE. THE REASON WHY THE B 4506 03:12:21,200 --> 03:12:22,680 SCORES, IT IS HARD CONCEPT AND 4507 03:12:22,680 --> 03:12:25,000 TAKES A LOT OF MATHEMATICAL 4508 03:12:25,000 --> 03:12:28,960 CALCULATIONS. SO PEOPLE LOOK FOR 4509 03:12:28,960 --> 03:12:30,600 SIMPLER THINGS. SO LET ME JUST 4510 03:12:30,600 --> 03:12:33,040 GET A LITTLE BIT OF A PRIMER ON 4511 03:12:33,040 --> 03:12:35,640 STATISTICS. SO THERE'S 4512 03:12:35,640 --> 03:12:36,800 STATISTICS ARE JUST COMPARISONS 4513 03:12:36,800 --> 03:12:39,120 AND THERE'S LOT OF USE AND 4514 03:12:39,120 --> 03:12:42,120 MISUSE OF THAT. AND I'M NOT SURE 4515 03:12:42,120 --> 03:12:45,840 IF YOU STUDY AMERICAN LITERATURE 4516 03:12:45,840 --> 03:12:46,360 BUT MARC TWAIN HAS BEEN 4517 03:12:46,360 --> 03:12:47,400 DESCRIBED TO SAY THERE'S THREE 4518 03:12:47,400 --> 03:12:50,960 KINDS OF LIES. THERE'S LIES, 4519 03:12:50,960 --> 03:12:52,520 DATA LIES AND STATISTICS. IF YOU 4520 03:12:52,520 --> 03:12:54,320 LOOK FOR THAT, THERE'S ENGLISH 4521 03:12:54,320 --> 03:12:55,360 PERSON THAT FIRST TALKED ABOUT 4522 03:12:55,360 --> 03:12:57,920 THAT. SO YOU NEED TO HAVE RULES 4523 03:12:57,920 --> 03:12:59,280 OF THUMB ABOUT STATISTICS AND 4524 03:12:59,280 --> 03:13:01,560 SIMPLE MEASURES FOR THE ASSAY 4525 03:13:01,560 --> 03:13:03,280 ROBUSTNESS. FIRST IN TERMS OF 4526 03:13:03,280 --> 03:13:05,160 WHAT IS THE STATISTICS? HERE IS 4527 03:13:05,160 --> 03:13:08,080 THE FORMULA DEFINITION FROM THIS 4528 03:13:08,080 --> 03:13:10,520 BOOK THAT I ALWAYS CITE ON THE 4529 03:13:10,520 --> 03:13:13,560 BOTTOM OF ALL MY SLIDES. IT IS A 4530 03:13:13,560 --> 03:13:14,880 COLLECTION OF METHODS FROM 4531 03:13:14,880 --> 03:13:18,640 PLANNING YOUR EXPERIMENTS TO 4532 03:13:18,640 --> 03:13:20,320 OBTAIN AND ORGANIZING YOUR DATA 4533 03:13:20,320 --> 03:13:23,480 SO SUMMARIZE RESULTS SO THAT YOU 4534 03:13:23,480 --> 03:13:24,880 CAN SAY YOU CAN DRAW CONCLUSIONS 4535 03:13:24,880 --> 03:13:27,120 THAT'S WHAT YOU ARE MEASURING IS 4536 03:13:27,120 --> 03:13:27,960 DIFFERENT HAHN THE NOISE WITH 4537 03:13:27,960 --> 03:13:30,680 THE BACKGROUND OR UNPERTURBED 4538 03:13:30,680 --> 03:13:33,640 SYSTEMS. YOU BASICALLY MAKE A 4539 03:13:33,640 --> 03:13:36,080 BUNCH OF MEASUREMENTS OR MAKE 4540 03:13:36,080 --> 03:13:37,360 BUNCH OF DETERMINATIONS AND EACH 4541 03:13:37,360 --> 03:13:39,960 OF THOSE MEASUREMENTS COMPRISE 4542 03:13:39,960 --> 03:13:44,200 SOMETHING CALLED POPULATION 4543 03:13:44,200 --> 03:13:46,400 ENSEMBLE OF MEASUREMENTS SO 4544 03:13:46,400 --> 03:13:47,880 STATISTICS HERE WOULD BE THERE 4545 03:13:47,880 --> 03:13:49,280 IS ABOUT A HUNDRED OF US HERE, 4546 03:13:49,280 --> 03:13:52,080 WE EACH HAVE A HEIGHT AND IF YOU 4547 03:13:52,080 --> 03:13:53,240 POP ON THE TABLE THAT'S YOUR 4548 03:13:53,240 --> 03:13:56,000 POPULATION METRIC. CENSUS IS 4549 03:13:56,000 --> 03:13:57,560 WHEN YOU HAVE A REALLY LARGE 4550 03:13:57,560 --> 03:13:58,480 POPULATION YOU CAN'T 4551 03:13:58,480 --> 03:14:00,080 INDIVIDUALLY TEST EVERYONE BUT 4552 03:14:00,080 --> 03:14:03,120 YOU DO SAMPLE GROUPS AND BLEND 4553 03:14:03,120 --> 03:14:04,520 THEM TOGETHER AND YOU CAN THEN 4554 03:14:04,520 --> 03:14:05,640 ASSUME THAT REPRESENTS THE WHOLE 4555 03:14:05,640 --> 03:14:07,080 POPULATION. THERE'S WAYS OF 4556 03:14:07,080 --> 03:14:10,720 TESTING FOR THAT. (INAUDIBLE) IS 4557 03:14:10,720 --> 03:14:12,600 A SUB COLLECTION OF ELEMENTS 4558 03:14:12,600 --> 03:14:14,360 FROM THAT POPULATION. AND THE 4559 03:14:14,360 --> 03:14:17,080 PARAMETER IS THE ACTUAL 4560 03:14:17,080 --> 03:14:18,000 NUMERICAL MEASUREMENT YOU MAKE 4561 03:14:18,000 --> 03:14:22,760 TO CHARACTERIZE THAT POPULATION. 4562 03:14:22,760 --> 03:14:24,760 A STATISTIC IS NUMERICAL 4563 03:14:24,760 --> 03:14:26,320 MEASUREMENT THAT DESCRIBES 4564 03:14:26,320 --> 03:14:28,480 CHARACTERISTIC OF THE SAMPLE. 4565 03:14:28,480 --> 03:14:30,560 THERE'S TWO KINDS OF DATA, WE 4566 03:14:30,560 --> 03:14:33,200 MOSTLY DEAL WITH QUANTITATIVE 4567 03:14:33,200 --> 03:14:37,080 DATA FOR INHIBITION OF CERTAIN 4568 03:14:37,080 --> 03:14:40,880 VALUE. BUT STATISTICS ON 4569 03:14:40,880 --> 03:14:42,440 CATEGORICAL DATA, A DIFFERENT 4570 03:14:42,440 --> 03:14:43,600 COLOR OR SOMETHING LARGER OR 4571 03:14:43,600 --> 03:14:51,520 LESS THAN SOMETHING ELSE. HERE 4572 03:14:51,520 --> 03:14:52,640 ARE EXAMPLES OF MISUSE OF 4573 03:14:52,640 --> 03:14:54,880 STATISTICS THE USE OF STATISTICS 4574 03:14:54,880 --> 03:14:59,040 IS TO TALK ABOUT QUALITY 4575 03:14:59,040 --> 03:15:00,640 CONTROL. IF SOMETHING HAS TIGHT 4576 03:15:00,640 --> 03:15:04,200 QUALITY IT HAS VERY LITTLE 4577 03:15:04,200 --> 03:15:06,040 VARIANCE, DOESN'T CHANGE DAY TO 4578 03:15:06,040 --> 03:15:08,920 DAY. YOU WANT TO BE ABLE TO GIVE 4579 03:15:08,920 --> 03:15:14,960 A NUMBER THAT ESTIMATES THAT 4580 03:15:14,960 --> 03:15:18,880 STATISTIC THEY HAVE SO SAY HOW 4581 03:15:18,880 --> 03:15:20,600 MANY PEOPLE IT AFFECTS WHAT IS 4582 03:15:20,600 --> 03:15:24,160 MY ASSURETY OF THAT THING. SO 4583 03:15:24,160 --> 03:15:26,840 POPULATION YOU SAY SOMETHING YOU 4584 03:15:26,840 --> 03:15:29,760 HAVE TO COUNT THEM AND CHECK ON 4585 03:15:29,760 --> 03:15:31,440 TRENDS. BASE STATISTICS WHEN 4586 03:15:31,440 --> 03:15:33,240 THEY SET A SPEED LIMIT IT IS NOT 4587 03:15:33,240 --> 03:15:36,280 JUST COMPLETELY ARBITRARY THEY 4588 03:15:36,280 --> 03:15:38,120 SAY HOW MANY ACCIDENTS AT THIS 4589 03:15:38,120 --> 03:15:39,600 SPEED THEY THINK IT IS NOT SAFE 4590 03:15:39,600 --> 03:15:42,200 THEY REDUCE IT SO THIS STATISTIC 4591 03:15:42,200 --> 03:15:44,440 THERE IS NUMBER OF ACCIDENTS PER 4592 03:15:44,440 --> 03:15:48,160 NUMBER OF MILES DRIVEN. SO YOU 4593 03:15:48,160 --> 03:15:50,560 HAVE TO ANALYZE. ESPECIALLY IN 4594 03:15:50,560 --> 03:15:52,160 OUR INDUSTRY WE HAVE TO BE ABLE 4595 03:15:52,160 --> 03:15:57,000 TO RECOGNIZE AND WHETHER STORY 4596 03:15:57,000 --> 03:15:58,360 STATISTICS. SO THERE ARE LOTS 4597 03:15:58,360 --> 03:16:00,200 OF ABUSES. SOME ABUSE IS LIKE 4598 03:16:00,200 --> 03:16:02,920 YOU HAVE SMALL OR BAD SAMPLE AND 4599 03:16:02,920 --> 03:16:06,120 IT IS KIND OF LINE, YOU CAN SAY 4600 03:16:06,120 --> 03:16:08,040 SEVEN OUT OF TEN DENTISTS. 4601 03:16:08,040 --> 03:16:12,880 SOUND LIKE 70% BUT THEY ONLY 4602 03:16:12,880 --> 03:16:15,000 ENTER TEN DENTISTS SO THAT IS A 4603 03:16:15,000 --> 03:16:17,240 SMALL STATISTIC OR EITHER YET 4604 03:16:17,240 --> 03:16:18,760 YOU FIND THEY WERE RESPOND CORED 4605 03:16:18,760 --> 03:16:21,040 BY DRUG COMPANIES SO THEIR PAID 4606 03:16:21,040 --> 03:16:23,080 SO THAT IS A BIAS STATISTIC. THE 4607 03:16:23,080 --> 03:16:25,000 OTHER ONE THAT IS COMMON, THIS 4608 03:16:25,000 --> 03:16:27,200 IS REFERS TO SIGNIFICANT FIGURES 4609 03:16:27,200 --> 03:16:29,360 AS WELL. THERE IS PRECISE 4610 03:16:29,360 --> 03:16:30,880 NUMBERS APPARENTLY PRECISE WHICH 4611 03:16:30,880 --> 03:16:34,400 GIVES YOU A FALSE SENSE OF 4612 03:16:34,400 --> 03:16:39,440 ACCURACY. LIKE $37,700. IS THAT 4613 03:16:39,440 --> 03:16:41,000 REALLY, IS IT REALLY BETTER TO 4614 03:16:41,000 --> 03:16:44,000 GIVE THEM ALL THESE ADDITIONAL 4615 03:16:44,000 --> 03:16:45,280 VIEW -- UNLESS YOU ARE COUNTING 4616 03:16:45,280 --> 03:16:46,480 THAT'S NOT IMPORTANT BECAUSE YOU 4617 03:16:46,480 --> 03:16:49,240 TRY TO SAY SOMETHING MORE 4618 03:16:49,240 --> 03:16:51,280 IMPORTANT. I HAVE BEEN IN 4619 03:16:51,280 --> 03:16:53,000 MEETINGS WITH VERY SERIOUS 4620 03:16:53,000 --> 03:16:55,640 PEOPLE WILL ARGUE WHAT IS LIKE 4621 03:16:55,640 --> 03:17:02,360 IC 50 OF 3.25 VERSUS 3.21. I ASK 4622 03:17:02,360 --> 03:17:03,800 HOW MANY SIGNIFICANT FIGURES 4623 03:17:03,800 --> 03:17:06,400 THEY RUN THAT COMPOUND OUT TO, 4624 03:17:06,400 --> 03:17:08,920 ATELL ME TWO MILLIGRAMS PLUS OR 4625 03:17:08,920 --> 03:17:10,640 MINUS ONE MILLIGRAM. SO THERE IS 4626 03:17:10,640 --> 03:17:13,240 ONLY ONE SIGNIFICANT FIGURE IN 4627 03:17:13,240 --> 03:17:16,320 THAT ENTIRE THING. SO ALWAYS ASK 4628 03:17:16,320 --> 03:17:19,480 THAT. THE ONLY TIME I USE 4629 03:17:19,480 --> 03:17:21,360 BALANCES YOU CAN MEASURE THINGS 4630 03:17:21,360 --> 03:17:23,520 TO THREE SIGNIFICANT FIGURES, 4631 03:17:23,520 --> 03:17:26,720 ACTUALLY FOUR IN TERMS OF 4632 03:17:26,720 --> 03:17:29,640 HUNDREDS OF MILLIGRAM. BUT IF 4633 03:17:29,640 --> 03:17:31,200 YOU PUT A FINGER PRINT YOU 4634 03:17:31,200 --> 03:17:33,760 CHANGE THE WAY SO JUST HAVE TO 4635 03:17:33,760 --> 03:17:35,320 DEAL WIT. THE OTHER ONE THAT'S 4636 03:17:35,320 --> 03:17:37,000 POPULAR IN JOURNALS IS SOMETHING 4637 03:17:37,000 --> 03:17:41,200 LIKE ON THE RIGHT. THIS IS THE 4638 03:17:41,200 --> 03:17:44,120 SAME DATA BUT PRESENTED THIS. SO 4639 03:17:44,120 --> 03:17:45,480 IF YOU ARE NOT CAREFUL YOU THINK 4640 03:17:45,480 --> 03:17:56,000 THAT IS A BIG CHANGE. IT IS A 4641 03:18:01,880 --> 03:18:02,640 SMALL PERCENT DIFFERENCE. THIS 4642 03:18:02,640 --> 03:18:05,200 IS THE THING TO WORRY ABOUT AND 4643 03:18:05,200 --> 03:18:06,720 IF YOU READ JOURNAL ARTICLES YOU 4644 03:18:06,720 --> 03:18:12,640 WILL SEE THIS. FOR NOW -- 4645 03:18:12,640 --> 03:18:15,240 ANALYSIS OF HDS DATA WE DO HAVE 4646 03:18:15,240 --> 03:18:18,960 TO PREPROCESS DATA, THAT IS 4647 03:18:18,960 --> 03:18:21,440 NORMALIZATION. BUT BEFORE 4648 03:18:21,440 --> 03:18:22,640 NORMALIZATION LOOK AT THE RAW 4649 03:18:22,640 --> 03:18:27,800 DATA. TO SEE IF THERE IS SOME 4650 03:18:27,800 --> 03:18:29,840 BIAS TO THAT. WITH THE KINDS OF 4651 03:18:29,840 --> 03:18:31,840 CORRECTIONS THE REASON TO DONOR 4652 03:18:31,840 --> 03:18:35,280 MALLIZATION IS TO CHECK PLATE BY 4653 03:18:35,280 --> 03:18:35,960 PLATED BUT SOME OF THE STANDARD 4654 03:18:35,960 --> 03:18:38,360 CONTROLS IN TERMS OF CORRECTING 4655 03:18:38,360 --> 03:18:40,760 FOR SYSTEMATIC ERRORS ARE 4656 03:18:40,760 --> 03:18:42,400 EXPERIMENTAL ERRORS TO SEE IF 4657 03:18:42,400 --> 03:18:48,120 THERE'S OUTLIERS IN THE CONTROL, 4658 03:18:48,120 --> 03:18:49,160 WHETHER THERE'S POSITIONAL 4659 03:18:49,160 --> 03:18:51,240 EFFECTS THE WAY YOU CHECK 4660 03:18:51,240 --> 03:18:52,680 POSITIONAL AFFECTS IF YOU TAKE 4661 03:18:52,680 --> 03:18:54,520 THE PLATE AND READ IT AND FLIP 4662 03:18:54,520 --> 03:18:57,680 360 DEGREES AND SEE IF LOW WELLS 4663 03:18:57,680 --> 03:19:01,720 TRANSPOSE WITH THE GEOMETRY 4664 03:19:01,720 --> 03:19:04,200 VERSUS THE ACTUAL PLATE ITSELF. 4665 03:19:04,200 --> 03:19:09,880 THEN YOU WANT TO LOOK AT CONTROL 4666 03:19:09,880 --> 03:19:12,080 WELLS AND OVERALL PLATE AND SEE 4667 03:19:12,080 --> 03:19:15,160 IF IT DRIFTS. ALL THIS WILL 4668 03:19:15,160 --> 03:19:19,080 EFFECT HOW YOU SELECT. RIGHT? 4669 03:19:19,080 --> 03:19:23,840 YOU WANT TO GET RID OF THOSE TO 4670 03:19:23,840 --> 03:19:26,400 ANALYSIS DISTRIBUTION. SO THE 4671 03:19:26,400 --> 03:19:28,720 VARIABILITY REPLICATE CONTROLS 4672 03:19:28,720 --> 03:19:35,080 IS USED TO ESTIMATE THE 4673 03:19:35,080 --> 03:19:36,200 VARIABILITY. I WANT TO 4674 03:19:36,200 --> 03:19:37,800 EMPHASIZE, WHEN YOU ARE STARTING 4675 03:19:37,800 --> 03:19:40,120 ASSAY YOU CAN TAKE WHOLE PLATE, 4676 03:19:40,120 --> 03:19:44,120 HIGH AND LOW CONTROL AND YOU CAN 4677 03:19:44,120 --> 03:19:45,080 CALCULATE THIS VARIABILITY FROM 4678 03:19:45,080 --> 03:19:47,960 THAT. WHEN YOU SCREEN YOU ONLY 4679 03:19:47,960 --> 03:19:49,400 HAVE CERTAIN WELL THAT ARE 4680 03:19:49,400 --> 03:19:52,400 CONTROLS AND YOU SCREEN ACTUAL 4681 03:19:52,400 --> 03:19:54,880 COMPOUND. BUT FOR MOST ASSAYS 4682 03:19:54,880 --> 03:19:59,320 AND TYPES OF WAY WE SCREEN, IT 4683 03:19:59,320 --> 03:20:02,280 TURNS OUT THAT VARIABILITY OF 4684 03:20:02,280 --> 03:20:03,360 REPLICATE WITHOUT COMPOUNDS YOU 4685 03:20:03,360 --> 03:20:07,960 ARE TESTING, IS OFTEN THE 4686 03:20:07,960 --> 03:20:09,680 OVERALL VARIABILITY, OF THOSE 4687 03:20:09,680 --> 03:20:10,920 CONTROLS, DO REFLECT THE 4688 03:20:10,920 --> 03:20:17,840 VARIABILITY OF YOUR SCREEN. WAS 4689 03:20:17,840 --> 03:20:20,640 BECAUSE MOST ARE INACTIVE. 99 4690 03:20:20,640 --> 03:20:23,760 ARE HITS AND THE REST ARE 4691 03:20:23,760 --> 03:20:24,680 INACTIVE COMPOUNDS. THAT'S WHAT 4692 03:20:24,680 --> 03:20:25,960 THE LAST LINE MEANS SO YOU CAN 4693 03:20:25,960 --> 03:20:28,680 LOOK AT RANDOM ERRORS VARIABLE 4694 03:20:28,680 --> 03:20:31,840 AGAINST ALL COMPOUNDS IN A PLATE 4695 03:20:31,840 --> 03:20:33,800 BECAUSE MOST OF THE COMPOUNDS 4696 03:20:33,800 --> 03:20:37,680 AREN'T ACTIVE AND INACTIVE 4697 03:20:37,680 --> 03:20:39,680 COMPOUNDS JUST REFLECT BY RANDOM 4698 03:20:39,680 --> 03:20:42,680 ERROR SO THAT SHOULD BE 4699 03:20:42,680 --> 03:20:46,120 DISTRIBUTED OR GALSION. SO THIS 4700 03:20:46,120 --> 03:20:48,720 IS AN EXAMPLE OF 1530 WELL PLATE 4701 03:20:48,720 --> 03:20:51,040 I HIGHLIGHT BASICALLY IF YOU 4702 03:20:51,040 --> 03:20:52,960 LOOK AT MOST 1536 WELL PLACE, 4703 03:20:52,960 --> 03:20:55,960 LIKE IN OUR LAB WE TEND TO PUT 4704 03:20:55,960 --> 03:20:57,560 MORE HIGHER CONTROLS THAN LOWER 4705 03:20:57,560 --> 03:20:59,080 CONTROLS BECAUSE THERE IS MORE 4706 03:20:59,080 --> 03:21:01,000 VARIANTS IN THE HIGH CONTROLS, 4707 03:21:01,000 --> 03:21:02,160 HIGH SIGNAL CONTROLS. SO WE DO 4708 03:21:02,160 --> 03:21:05,840 THAT. IN OUR CASE, MANY OF OUR 4709 03:21:05,840 --> 03:21:08,840 DISPENSERS THEY WORK IN A 4710 03:21:08,840 --> 03:21:10,480 CERTAIN FASHION SO YOU HAVE A 4711 03:21:10,480 --> 03:21:12,080 CLOGGED TIP YOU CAN SEE SPREAD 4712 03:21:12,080 --> 03:21:13,280 QUICKLY. SO THESE ARE OBVIOUS 4713 03:21:13,280 --> 03:21:16,080 THINGS YOU CAN ELIMINATE THOSE 4714 03:21:16,080 --> 03:21:17,360 DATA FROM ANALYSIS OR TRY TO 4715 03:21:17,360 --> 03:21:22,160 CORRECT THEM. WERE AVERAGE 4716 03:21:22,160 --> 03:21:24,640 CORRECTION ALGORITHM. ONE OF THE 4717 03:21:24,640 --> 03:21:27,040 REASON I ACTUALLY SHOW THIS 4718 03:21:27,040 --> 03:21:30,200 PARTICULAR EXAMPLE IS ONE WAY TO 4719 03:21:30,200 --> 03:21:32,200 PLOT TO MAKE SURE ASSAY IS 4720 03:21:32,200 --> 03:21:37,840 WORKING WELL IS LOOK AT 4721 03:21:37,840 --> 03:21:39,480 DISTRIBUTION OF CURRENT 4722 03:21:39,480 --> 03:21:40,240 DEFLECTION, IF FLAT LIKE THIS, 4723 03:21:40,240 --> 03:21:41,240 PROBABLY A GOOD PLATE TESTIMONY 4724 03:21:41,240 --> 03:21:42,760 OTHER THING, IF THERE'S AN AGE 4725 03:21:42,760 --> 03:21:47,080 EFFECT ESPECIALLY 1536, THESE 4726 03:21:47,080 --> 03:21:49,640 PLACE TEND TO HEAT UP FASTER. 4727 03:21:49,640 --> 03:21:51,120 MOST CONSEQUENCE IS ON CELL 4728 03:21:51,120 --> 03:21:53,240 BASED ASSAYS WHERE THEY DRY OUT 4729 03:21:53,240 --> 03:21:55,800 FASTER. SO ONE PRACTICAL ASPECT 4730 03:21:55,800 --> 03:22:06,360 WHEN SCREENERS ARE APPROACHED BY 4731 03:22:07,600 --> 03:22:09,240 THEY WANT A NINE DAY ASSAY YOU 4732 03:22:09,240 --> 03:22:10,560 HAVE TO TELL THEM PRACTICALLY 4733 03:22:10,560 --> 03:22:14,120 YOU CAN'T DO MORE THAN A 5A 4734 03:22:14,120 --> 03:22:15,920 ASSAY WITHOUT HEROIC MEASURES. 4735 03:22:15,920 --> 03:22:19,320 THAT'S LIMITATION. AND THE 4736 03:22:19,320 --> 03:22:21,120 LIMITATION IS ADDRESSED PROBABLY 4737 03:22:21,120 --> 03:22:26,120 BY GOING TO 3D FOUR WELL PLACE. 4738 03:22:26,120 --> 03:22:30,160 384 DOES WORK IN MANY FORMATS. 4739 03:22:30,160 --> 03:22:32,960 THIS TABLE I PUT TOGETHER MAKES 4740 03:22:32,960 --> 03:22:36,400 A POINT THAT NATHAN MIGHT HAVE 4741 03:22:36,400 --> 03:22:38,880 SHOWED TO BUT 96 WELLS AND 36 4742 03:22:38,880 --> 03:22:41,960 WELLS, THE BIG SURPRISE THING IS 4743 03:22:41,960 --> 03:22:43,840 MOST 1536 WELLS TEND TO BE LONG 4744 03:22:43,840 --> 03:22:48,080 TALL COLUMNS AND 384 -- 96 WELLS 4745 03:22:48,080 --> 03:22:49,080 TEND PLAID FLAT WIDE DISCS OR 4746 03:22:49,080 --> 03:22:50,320 BARRELS SO WHAT IT MEANS IS IF 4747 03:22:50,320 --> 03:22:53,960 YOU HAVE SOMETHING THAT IS 4748 03:22:53,960 --> 03:22:57,600 EVAPORATES IN TERMS OF OR TRYING 4749 03:22:57,600 --> 03:22:59,360 TO GET AREA EXCHANGE, THE 4750 03:22:59,360 --> 03:23:01,960 SURFACE AREA OF O MENISCUS IS 4751 03:23:01,960 --> 03:23:04,360 SMALLER RELATIVE TO CONTAIN FOR 4752 03:23:04,360 --> 03:23:08,160 1536. SO 384 AIR RATES WELL, 4753 03:23:08,160 --> 03:23:10,640 1536 STRUGGLES THEY DON'T GROW 4754 03:23:10,640 --> 03:23:15,400 WELL HOWEVER THE VOLUME OF THE 4755 03:23:15,400 --> 03:23:19,720 THAT SURFACE AREA VOLUME CONTEXT 4756 03:23:19,720 --> 03:23:24,760 IN A 1536 IS HIGHER MORE SURFACE 4757 03:23:24,760 --> 03:23:26,680 CONTACT IN THE BAR SO THAT HAS 4758 03:23:26,680 --> 03:23:29,360 COUPLE OF EFFECTS. IF YOU DO SO 4759 03:23:29,360 --> 03:23:32,680 LIKE WASH ASSAY THAT RELY ON 4760 03:23:32,680 --> 03:23:35,000 ADHERENCE OF YOUR TARGET TO 4761 03:23:35,000 --> 03:23:37,040 WELLS, 1536 SCALES BETTER. IT 4762 03:23:37,040 --> 03:23:39,560 ALSO MEANS ANY TYPE OF STICKY 4763 03:23:39,560 --> 03:23:40,760 COMPOUNDS ARE EXACERBATED IN 4764 03:23:40,760 --> 03:23:43,800 TERMS OF DEPLETION WITH 1536 4765 03:23:43,800 --> 03:23:45,320 PLATE. THAT'S ONE PRACTICAL 4766 03:23:45,320 --> 03:23:49,000 ASPECT. SO THIS IS HOW YOU LOOK 4767 03:23:49,000 --> 03:23:51,240 AT SYSTEMATIC BASED ERRORS LOOK 4768 03:23:51,240 --> 03:23:52,840 AT THINGS LIKE IF YOU PLOT 4769 03:23:52,840 --> 03:23:57,360 THINGS BY COLUMNS, OR BY ROWS 4770 03:23:57,360 --> 03:24:01,080 YOU CAN SEE IF A COLUMN IS 4771 03:24:01,080 --> 03:24:02,320 ALWAYS HIGH VERSUS OTHER COLUMNS 4772 03:24:02,320 --> 03:24:05,840 SO THIS IS A COLUMN THAT MIGHT 4773 03:24:05,840 --> 03:24:06,880 BE EDGE EFFECT SO YOU WANT TO DO 4774 03:24:06,880 --> 03:24:12,760 THAT. YOU WANT TO SEE IF -- SO 4775 03:24:12,760 --> 03:24:15,000 SAME THING HERE THIS SYSTEMATIC 4776 03:24:15,000 --> 03:24:16,640 DRIFT EACH TIME OR HERE YOU ARE 4777 03:24:16,640 --> 03:24:19,760 SEEING WITH INCUBATION VERY 4778 03:24:19,760 --> 03:24:23,840 PATTERN DRIFT SO EVERY COLUMN IS 4779 03:24:23,840 --> 03:24:25,360 INCREASING IN TIME, YOU CAN PLOT 4780 03:24:25,360 --> 03:24:26,840 THIS WAY AND THIS GIVES YOU 4781 03:24:26,840 --> 03:24:28,680 CLEARER IDEAS IF THERE'S 4782 03:24:28,680 --> 03:24:31,680 SYSTEMATIC ERRORS. THE WAY YOU 4783 03:24:31,680 --> 03:24:33,760 DO THAT IS WAY YOU CORRECT IS 4784 03:24:33,760 --> 03:24:37,880 REDESIGN THE SAY, YOU GET RID OF 4785 03:24:37,880 --> 03:24:38,960 THE OUTSIDE WELLS OR IF YOU 4786 03:24:38,960 --> 03:24:40,800 CAN'T GET RID OF THEM YOU CAN 4787 03:24:40,800 --> 03:24:44,560 PUT SMOOTHING ALGORITHMS AND 4788 03:24:44,560 --> 03:24:46,600 TAKE THE AVERAGE SO WHEN YOU ARE 4789 03:24:46,600 --> 03:24:50,600 DONE WITH THAT, YOU ARE LEFT 4790 03:24:50,600 --> 03:24:54,120 WITH RANDOM ERROR. I'M GOING TO 4791 03:24:54,120 --> 03:24:58,240 JUMP HERE TO THIS THING WHICH 4792 03:24:58,240 --> 03:25:00,160 HAS BEEN TAUGHT TO ME BUT IT IS 4793 03:25:00,160 --> 03:25:02,280 A LITTLE COUNTER INTUITIVE. I 4794 03:25:02,280 --> 03:25:06,920 WILL WALK -- THIS IS A 4795 03:25:06,920 --> 03:25:13,680 DISTRIBUTION OF ALL THESE PEOPLE 4796 03:25:13,680 --> 03:25:15,960 ABOUT I PRESIDENT HOW MANY 4797 03:25:15,960 --> 03:25:18,960 PEOPLE -- 200 DIGITS SO TAKE 4798 03:25:18,960 --> 03:25:20,720 THIS SOCIAL SECURITY NUMBER OF 4799 03:25:20,720 --> 03:25:22,000 ANYONE RANDOMLY AND LOOK AT FOUR 4800 03:25:22,000 --> 03:25:28,400 OF THE THEM. RIGHT? FROM 0 TO #E 4801 03:25:28,400 --> 03:25:31,280 DISTRIBUTION IS IF YOU TAKE 200, 4802 03:25:31,280 --> 03:25:33,000 IT SHOULD BE ABOUT THERE'S NINE 4803 03:25:33,000 --> 03:25:36,600 RANDOM NUMBERS, EQUALLY 4804 03:25:36,600 --> 03:25:38,160 POSSIBLE. SO THEY DID THIS AND 4805 03:25:38,160 --> 03:25:39,440 THEN THIS IS THE DISTRIBUTION 4806 03:25:39,440 --> 03:25:43,680 YOU GET. IT IS RELATIVELY FLAT. 4807 03:25:43,680 --> 03:25:45,320 YOU CAN FORMALLY CALCULATE THE 4808 03:25:45,320 --> 03:25:47,760 MEAN OF THIS, THE MEAN IS FOUR 4809 03:25:47,760 --> 03:25:49,760 AND A HALF BECAUSE ONE TO -- IT 4810 03:25:49,760 --> 03:25:54,400 IS 0 TO 9. SO THAT MAKES SENSE. 4811 03:25:54,400 --> 03:25:56,320 THE SIGNIFICANT FIGURES 4812 03:25:56,320 --> 03:25:57,160 REPRESENTS JUST COUNTING 4813 03:25:57,160 --> 03:25:58,280 STATISTICS WHEN YOU DO IT. WHAT 4814 03:25:58,280 --> 03:26:02,600 IS INTERESTING IS IF YOU FAKE 4815 03:26:02,600 --> 03:26:03,280 NOT THE ACTUAL DISTRIBUTION 4816 03:26:03,280 --> 03:26:08,280 WHICH IS FLAT, BUT CALCULATE THE 4817 03:26:08,280 --> 03:26:09,280 MEANS OF THE DISTRIBUTION -- IF 4818 03:26:09,280 --> 03:26:11,480 YOU TAKE ALL THESE SAMPLES AND 4819 03:26:11,480 --> 03:26:13,320 CALIFORNIA COW LATE THE MEAN OF 4820 03:26:13,320 --> 03:26:14,960 FOUR SIGNATURES, AND PLOT, SO 4821 03:26:14,960 --> 03:26:20,320 HERE IS THE MEAN OF FOUR DIGITS 4822 03:26:20,320 --> 03:26:21,160 AND LOUT THAT NUMBER, THE MEAN 4823 03:26:21,160 --> 03:26:23,520 EVERY FOUR AND PLOT THE 4824 03:26:23,520 --> 03:26:24,160 DISTRIBUTION OF THE MEANS YOU 4825 03:26:24,160 --> 03:26:27,720 SEE HOW IT LOOKS SORT OF LIKE A 4826 03:26:27,720 --> 03:26:30,320 NORMAL DISTRIBUTION. IF YOU 4827 03:26:30,320 --> 03:26:33,120 ACTUALLY INCREASE THAT NUMBER OF 4828 03:26:33,120 --> 03:26:34,920 SAMPLES YOU DO TO CALCULATE THE 4829 03:26:34,920 --> 03:26:37,120 MEAN YOU MAKE THAT N VALUE 4830 03:26:37,120 --> 03:26:41,640 LARGER IT BECOMES MORE GALSION. 4831 03:26:41,640 --> 03:26:43,160 SO WHY IS THIS IMPORTANT? IT IS 4832 03:26:43,160 --> 03:26:44,360 COUNTER INTUITIVE THAT THIS 4833 03:26:44,360 --> 03:26:46,560 SHOULD BE BUT YOU CAN DO IT MANY 4834 03:26:46,560 --> 03:26:47,560 THINGS. SHOW YOU IN THE NEXT 4835 03:26:47,560 --> 03:26:49,880 SLIDE. THE MEAN STAYS THE SAME 4836 03:26:49,880 --> 03:26:50,800 FROM THE FIRST DISTRIBUTION -- 4837 03:26:50,800 --> 03:26:53,920 FROM THE POPULATION DISTRIBUTION 4838 03:26:53,920 --> 03:26:57,680 TO MEAN SAMPLE DISTRIBUTION THAT 4839 03:26:57,680 --> 03:26:58,920 IS CONSTANT. WHAT IS INTERESTING 4840 03:26:58,920 --> 03:27:01,000 IS THIS -- AS YOU GO TO 4841 03:27:01,000 --> 03:27:01,920 POPULATION DISTRIBUTION OF 4842 03:27:01,920 --> 03:27:03,640 POPULATION MEANS,S THAT STANDARD 4843 03:27:03,640 --> 03:27:06,040 DEVIATION OF MEAN IS SMALLER 4844 03:27:06,040 --> 03:27:08,480 THAN THE STANDARD DEVIATION OF 4845 03:27:08,480 --> 03:27:09,800 POPULATION. IT IS DIFFERENT BY 4846 03:27:09,800 --> 03:27:12,600 REDUCING IT BY THE NUMBER -- 4847 03:27:12,600 --> 03:27:15,360 SCROLL TO THE EN END. THIS IS TT 4848 03:27:15,360 --> 03:27:19,120 STANDARD ERROR OF MEAN YOU 4849 03:27:19,120 --> 03:27:23,520 ALWAYS SEE. WHAT IS FUNNY ABOUT 4850 03:27:23,520 --> 03:27:25,640 THIS THING IS IF YOU APPLY THIS 4851 03:27:25,640 --> 03:27:27,520 TO ANY DISTRIBUTION WHETHER 4852 03:27:27,520 --> 03:27:32,320 UNIFORM LIKE THE FIRST CASE, 4853 03:27:32,320 --> 03:27:35,680 TRUE GALSION DISTRIBUTION, OR A 4854 03:27:35,680 --> 03:27:38,280 SKEWED DISTRIBUTION WHETHER 4855 03:27:38,280 --> 03:27:40,280 NORMAL UNIFORM OR SKEWED, 4856 03:27:40,280 --> 03:27:42,720 WHENEVER YOU TAKE THE SAMPLE 4857 03:27:42,720 --> 03:27:44,720 MEAN, IT BECOMES MORE NORMA NORS 4858 03:27:44,720 --> 03:27:46,320 THE LARGER NUMBER OF SAMPLES YOU 4859 03:27:46,320 --> 03:27:52,080 PUT IN THAT MEAN, MORE NORMAL IT 4860 03:27:52,080 --> 03:27:54,280 GETS. THIS IS IMPORTANT FOR THE 4861 03:27:54,280 --> 03:27:55,560 REASON THAT THE MEAN ALWAYS 4862 03:27:55,560 --> 03:27:57,040 STAYS THE SAME AND SOMETIMES YOU 4863 03:27:57,040 --> 03:27:58,520 DON'T KNOW EVEN WITH THE 4864 03:27:58,520 --> 03:27:59,880 DISTRIBUTION YOU HAVE TO STILL 4865 03:27:59,880 --> 03:28:03,480 MAKE JUDGMENTS ON WHAT IS 4866 03:28:03,480 --> 03:28:06,360 PROBABLE. SO AGAIN THE MEAN 4867 03:28:06,360 --> 03:28:07,840 APPROACHES THE TRUE MEAN, 4868 03:28:07,840 --> 03:28:09,320 STANDARD DEVIATION IS BECOMES A 4869 03:28:09,320 --> 03:28:12,640 STANDARD ERROR OF THE MEANS WHEN 4870 03:28:12,640 --> 03:28:16,760 YOU GET LARGER POPULATIONS THEY 4871 03:28:16,760 --> 03:28:19,960 BECOME GALLION. WHY THIS SHOULD 4872 03:28:19,960 --> 03:28:22,880 MAKE SENSE IS WHILE ALL OF US 4873 03:28:22,880 --> 03:28:24,120 HAVE LIKE DIFFERENT WEIGHTS, IF 4874 03:28:24,120 --> 03:28:26,200 WE TAKE THE MEAN OF OUR WEIGHT, 4875 03:28:26,200 --> 03:28:29,560 IT IS KIND OF LIKE SAYING HOW 4876 03:28:29,560 --> 03:28:30,720 DIFFERENT ARE ARCH OF 30 PEOPLE 4877 03:28:30,720 --> 03:28:32,360 VERSUS JUST ONE OR TWO PEOPLE? 4878 03:28:32,360 --> 03:28:35,800 SO THAT YOU CAN SAY CHANCE OF 4879 03:28:35,800 --> 03:28:38,040 FINDING SOMEONE TALLEST IN THE 4880 03:28:38,040 --> 03:28:39,400 GROUP IS PROBABLY IF SOMEONE IS 4881 03:28:39,400 --> 03:28:41,360 TALL VERSUS THE REST, THAT IS A 4882 03:28:41,360 --> 03:28:43,360 BIG VARIANCE BUT IF I AVERAGE 4883 03:28:43,360 --> 03:28:47,000 THAT PERSON WHO IS TALL AMONG 30 4884 03:28:47,000 --> 03:28:48,720 OTHER PEOPLE, THE IMPACT OF 4885 03:28:48,720 --> 03:28:51,600 THEIR VARIANCE IS MINIMIZED. SO 4886 03:28:51,600 --> 03:28:54,400 THE REASON THIS IS IMPORTANT, 4887 03:28:54,400 --> 03:28:56,040 YOU CAN ACTUALLY ALWAYS COUNTER 4888 03:28:56,040 --> 03:28:59,320 THE SAMPLE MEAN AND ASSUME ITS 4889 03:28:59,320 --> 03:29:00,640 NORMAL AND THEN BACK CALCULATE 4890 03:29:00,640 --> 03:29:02,600 WHAT THE STANDARD DEVIATION OF 4891 03:29:02,600 --> 03:29:03,920 THE POPULATION WAS BECAUSE YOU 4892 03:29:03,920 --> 03:29:05,360 MOW NUMBER OF REPLICATES YOU DO, 4893 03:29:05,360 --> 03:29:06,920 YOU CAN GET STANDARD DEVIATION 4894 03:29:06,920 --> 03:29:11,800 OF YOUR SAMPLE MEANS. THEN YOU 4895 03:29:11,800 --> 03:29:13,960 CAN MULTIPLY IT BY THE END AND 4896 03:29:13,960 --> 03:29:14,680 GET ACTUAL DISTRIBUTION OF THE 4897 03:29:14,680 --> 03:29:19,840 POPULATION. IT IS CENTRAL TO LOT 4898 03:29:19,840 --> 03:29:24,760 OF STATISTICAL THEORY. SO WHAT 4899 03:29:24,760 --> 03:29:28,320 IS THE MEAN? WE USED ARITHMETIC 4900 03:29:28,320 --> 03:29:29,680 MEAN, THIS IS BASICALLY IF YOU 4901 03:29:29,680 --> 03:29:31,920 THINK ABOUT ANY DISTRIBUTION, 4902 03:29:31,920 --> 03:29:34,400 AND PLOT WEIGHTS IT IS THE 4903 03:29:34,400 --> 03:29:37,000 FULCRUM POINT TO BALANCE THIS 4904 03:29:37,000 --> 03:29:39,360 WHOLE DISTRIBUTION. THE 4905 03:29:39,360 --> 03:29:41,120 ARITHMETIC MEAN IS SUM OF ALL 4906 03:29:41,120 --> 03:29:43,560 INDIVIDUALS DIVIDE BY NUMBER OF 4907 03:29:43,560 --> 03:29:44,760 MEASUREMENTS. IT ACTUALLY IS 4908 03:29:44,760 --> 03:29:46,160 USEFUL ALL THE TIME, YOU CAN 4909 03:29:46,160 --> 03:29:48,360 ALWAYS CALCULATE IT. IT IS 4910 03:29:48,360 --> 03:29:52,360 AFFECTED BY EXTREMES. THE MOST 4911 03:29:52,360 --> 03:29:53,800 FAMILIAR WITH WHAT -- BUT THERE 4912 03:29:53,800 --> 03:29:55,400 ARE HOARE KINDS OF MEANS YOU CAN 4913 03:29:55,400 --> 03:29:57,120 CALCULATE IN TERMS OF CENTRAL 4914 03:29:57,120 --> 03:29:58,240 TENDENCY. ONE IS CALLED THE 4915 03:29:58,240 --> 03:30:00,520 MEDIAN. THAT IS IMPORTANT 4916 03:30:00,520 --> 03:30:02,840 BECAUSE BASICALLY WHAT IS THE 4917 03:30:02,840 --> 03:30:05,960 MIDDLE VALUE OF ANYTHING? THAT'S 4918 03:30:05,960 --> 03:30:07,040 INSENSITIVE TO REAL WIDE 4919 03:30:07,040 --> 03:30:11,120 DEVIATION BECAUSE MIDDLE VALUE. 4920 03:30:11,120 --> 03:30:13,000 THE MOST IS THE MOST FREQUENT 4921 03:30:13,000 --> 03:30:14,600 VALUE NOT SENSITIVE TO THE WIDE 4922 03:30:14,600 --> 03:30:17,160 RANGE BECAUSE IT IS CENTRAL OF 4923 03:30:17,160 --> 03:30:18,880 THE MOST COMMON VALU VALUE. ONET 4924 03:30:18,880 --> 03:30:20,600 IS AFFECTED BY EXTREMES IS MID 4925 03:30:20,600 --> 03:30:22,640 RANGE, AND THE WAY IT PRESENTS 4926 03:30:22,640 --> 03:30:24,840 IS IF YOU HAVE A NORMAL 4927 03:30:24,840 --> 03:30:25,680 DISTRIBUTION ALL THE MEANS ARE 4928 03:30:25,680 --> 03:30:29,200 THE SAME. BUT YOU CAN SEE IF 4929 03:30:29,200 --> 03:30:30,800 YOU HAVE A SKEWED DISTRIBUTION 4930 03:30:30,800 --> 03:30:34,880 TO THE LEFT, THE ACTUAL 4931 03:30:34,880 --> 03:30:36,080 ARITHMETIC MEAN IS HIGHER THAN 4932 03:30:36,080 --> 03:30:41,360 THE MODE. SO THESE ARE ONES THAT 4933 03:30:41,360 --> 03:30:44,800 PEOPLE DO CHECK ON. STANDARD 4934 03:30:44,800 --> 03:30:47,240 DEVIATION IS A MEASUREMENT OF 4935 03:30:47,240 --> 03:30:49,240 VARIATION. AND I THINK WE ALL 4936 03:30:49,240 --> 03:30:52,360 INTUITIVELY KNOW IF YOU HAVE 4937 03:30:52,360 --> 03:30:54,760 ONLY ONE SAMPLE, THEN IT HAS NO 4938 03:30:54,760 --> 03:30:55,680 STANDARD DEVIATION BECAUSE IT IS 4939 03:30:55,680 --> 03:30:59,320 A SINGLE ONE. IF YOU HAVE 4940 03:30:59,320 --> 03:31:01,840 DIFFERENT REPLICATES LIKE THIS, 4941 03:31:01,840 --> 03:31:03,200 THE DISTRIBUTED WEIRD, THIS 4942 03:31:03,200 --> 03:31:05,840 STANDARD DEVIATION SHOULD BE 4943 03:31:05,840 --> 03:31:09,120 FAIRLY LOW BUT HERE THERE IS 4944 03:31:09,120 --> 03:31:10,560 WIDE VARIANCE. SO STANDARD 4945 03:31:10,560 --> 03:31:11,600 DEVIATION IS SOMETIMES LARGER 4946 03:31:11,600 --> 03:31:15,280 THAN THE MEASUREMENT ITSELF. FOR 4947 03:31:15,280 --> 03:31:18,560 THE POPULATION, WHEN YOU GO 4948 03:31:18,560 --> 03:31:20,960 EPIFINTY THEY COLLAPSE TO SIGMA 4949 03:31:20,960 --> 03:31:22,640 THE MU AND TOTAL ENDS. IF YOU 4950 03:31:22,640 --> 03:31:26,360 HAVE A SAMPLE, ANYTHING SMALLER 4951 03:31:26,360 --> 03:31:27,280 THAN 30 YOU HAVE TO REDUCE 4952 03:31:27,280 --> 03:31:29,600 DEGREES OF FREEDOM BY ONE. SO 4953 03:31:29,600 --> 03:31:31,360 INSTEAD OF 30 SAMPLES YOU HAVE 4954 03:31:31,360 --> 03:31:35,640 DEGREE OF FREEDOM OF 29. SO 4955 03:31:35,640 --> 03:31:37,800 SCORES CLOSE TOGETHER WILL YIELD 4956 03:31:37,800 --> 03:31:39,040 SMALLER STANDARD DEVIATION. IT 4957 03:31:39,040 --> 03:31:40,840 IS INTUITIVE AND WE FORMALLY 4958 03:31:40,840 --> 03:31:41,880 HAVE DONE THAT. THE OTHER THING 4959 03:31:41,880 --> 03:31:44,240 THAT IS IMPORTANT FOR NORMAL 4960 03:31:44,240 --> 03:31:47,680 DISTRIBUTION IS TO UNDERSTAND # 4961 03:31:47,680 --> 03:31:49,720 9, 9568 RULE. WHAT IT MEANS IS 4962 03:31:49,720 --> 03:31:51,800 IN THREE UNITS THREE STANDARD 4963 03:31:51,800 --> 03:31:54,080 DEVIATION UNITS, 99% OF ALL THE 4964 03:31:54,080 --> 03:31:56,600 AREA UNDER THE CURVE IS 4965 03:31:56,600 --> 03:31:58,960 CONTAINED IN THAT. SO THAT IS 4966 03:31:58,960 --> 03:32:00,360 TRUE NORMAL BELL SHAPED 4967 03:32:00,360 --> 03:32:01,480 DISTRIBUTION IS. SO THAT IS HOW 4968 03:32:01,480 --> 03:32:04,080 WE ALWAYS SAY LIKE WE DO THREE 4969 03:32:04,080 --> 03:32:06,000 STANDARD DEVIATION THE MEAN. 4970 03:32:06,000 --> 03:32:08,120 THAT MEANS THE BULK OF THE 4971 03:32:08,120 --> 03:32:11,040 POPULATION ACTS LIKE NORMAL 4972 03:32:11,040 --> 03:32:11,600 DISTRIBUTION, IT IS ALL THE 4973 03:32:11,600 --> 03:32:13,320 SAME. WHAT IS DIFFERENT, ONE 4974 03:32:13,320 --> 03:32:14,560 PERCENT THAT IS DIFFERENT IS 4975 03:32:14,560 --> 03:32:15,720 THREE STANDARD DEVIATIONS AWAY 4976 03:32:15,720 --> 03:32:17,480 FROM THE MEAN. THAT IS HOW WE 4977 03:32:17,480 --> 03:32:22,040 CAME TO THAT NUMBER. THIS IS A 4978 03:32:22,040 --> 03:32:24,040 LITTLE BIT OF WHAT I INFERRED 4979 03:32:24,040 --> 03:32:27,000 FROM YEARS OF EXPERIENCE AND IT 4980 03:32:27,000 --> 03:32:28,160 IS NOT SOMETHING PEOPLE 4981 03:32:28,160 --> 03:32:30,840 EMPHASIZE BUT BRING TO YOUR 4982 03:32:30,840 --> 03:32:37,120 ATTENTION. ON THE TOP WHEN WE 4983 03:32:37,120 --> 03:32:40,200 SCREEN, THE REASON WHY IS HDS 4984 03:32:40,200 --> 03:32:42,400 DATA NORMALLY DISTRIBUTED AND 4985 03:32:42,400 --> 03:32:44,200 WHY TREAT IT LIKE THAT? I KIND 4986 03:32:44,200 --> 03:32:45,280 OF TALKED ABOUT IT SEVERAL 4987 03:32:45,280 --> 03:32:48,680 TIMES. ONE IS THAT RANDOM 4988 03:32:48,680 --> 03:32:50,000 COMPOUND COLLECTION 99% ARE NOT 4989 03:32:50,000 --> 03:32:53,960 ACTIVE. SO THEY SHOULD ACT LIKE 4990 03:32:53,960 --> 03:32:54,600 NOISE. THEY SHOULDN'T HAVE ANY 4991 03:32:54,600 --> 03:32:57,720 ACTIVITY. THE WAY TO MAKE IT 4992 03:32:57,720 --> 03:33:00,480 REAL, IF I DIDN'T -- WE DO THE 4993 03:33:00,480 --> 03:33:02,560 -- WHEN I WAS SUPERVISOR I COULD 4994 03:33:02,560 --> 03:33:05,240 HAVE TOLD ONE OF MY PEOPLE TO 4995 03:33:05,240 --> 03:33:07,480 SCREEN THIS LIBRARY BUT WHAT I 4996 03:33:07,480 --> 03:33:09,160 COULD HAVE DONE WAS TO FOOL THEM 4997 03:33:09,160 --> 03:33:11,160 COULD HAVE TAKEN A COMPOUND THAT 4998 03:33:11,160 --> 03:33:14,040 TEN MICROMOLAR IC 50 MADE 4999 03:33:14,040 --> 03:33:16,280 100,000 REPLY CASE AND SCREENED 5000 03:33:16,280 --> 03:33:18,720 THAT. WHAT THEY SHOULD FIND 5001 03:33:18,720 --> 03:33:21,960 USING THE STANDARD SCREEN IS 5002 03:33:21,960 --> 03:33:22,880 THAT THEY WILL FIND IF THEY 5003 03:33:22,880 --> 03:33:26,080 SCREEN TEN MICROMOLAR, ICD 10 5004 03:33:26,080 --> 03:33:26,800 MICROMOLAR THEY FIND 5005 03:33:26,800 --> 03:33:28,560 DISTRIBUTION THAT LOOKS NORMAL 5006 03:33:28,560 --> 03:33:30,480 BUT CERTAINTIED AT 50% 5007 03:33:30,480 --> 03:33:35,120 INHIBITION RATE. I BELABOR THIS 5008 03:33:35,120 --> 03:33:36,960 BECAUSE WHEN WE DO DOSE RESPONSE 5009 03:33:36,960 --> 03:33:38,520 CURVE THIS IS WHAT WE ARE DOING 5010 03:33:38,520 --> 03:33:41,080 TAKING REPLICATES TO SEE IF IT 5011 03:33:41,080 --> 03:33:45,360 IS EACH CONCENTRATION IN MY ALSO 5012 03:33:45,360 --> 03:33:47,760 GALSION AND IS THAT THE AVERAGE 5013 03:33:47,760 --> 03:33:49,360 THE OTHER REASON I BRING THIS UP 5014 03:33:49,360 --> 03:33:52,560 IS MANY TIMES YOU DO A SCREEN 5015 03:33:52,560 --> 03:33:54,400 AND YOU MANAGE, I SCREEN AND 5016 03:33:54,400 --> 03:33:55,640 CAN'T FIND ANYTHING, SCREEN 5017 03:33:55,640 --> 03:33:58,080 HIGHER CONCENTRATION YOU FIND 5018 03:33:58,080 --> 03:33:59,600 MORE THINGS. ACTUALLY IF YOU DO 5019 03:33:59,600 --> 03:34:02,720 THAT ALL YOU DO IS MOVING THE 5020 03:34:02,720 --> 03:34:05,920 POPULATION UP TO HIGHER AVERAGE 5021 03:34:05,920 --> 03:34:06,720 ACTIVITY AND COLLAPSING YOUR 5022 03:34:06,720 --> 03:34:09,360 WINDOW. THE REASON WHY REAL 5023 03:34:09,360 --> 03:34:12,120 SCREENS WORK, MOST OF THE 5024 03:34:12,120 --> 03:34:14,440 COMPOUNDS HUNDRED THOUSAND 5025 03:34:14,440 --> 03:34:17,920 COMPOUNDS, THAT YOU HAVE 5026 03:34:17,920 --> 03:34:22,160 POTENCIES LESS THAN ONE 5027 03:34:22,160 --> 03:34:24,440 MICROMOLAR. THE IDEAL SCREEN IS 5028 03:34:24,440 --> 03:34:28,160 WHERE THE CENTROID IS NEGATIVE 5029 03:34:28,160 --> 03:34:30,080 AND YOU HAVE SAMPLE LORING 5030 03:34:30,080 --> 03:34:31,560 SPREAD OF COMPOUNDS, SO WHAT I 5031 03:34:31,560 --> 03:34:32,880 LOOK FOR IN THE SCREEN IS 5032 03:34:32,880 --> 03:34:35,960 SOMETHING THAT LOOKS LIKE 5033 03:34:35,960 --> 03:34:38,880 CENTRAL TENDENCY IS GALLION BUT 5034 03:34:38,880 --> 03:34:40,560 THERE IS A SMEAR -- GALSION, I 5035 03:34:40,560 --> 03:34:41,760 LOOK AND THERE IS A SMEAR TO THE 5036 03:34:41,760 --> 03:34:43,240 RIGHT. THE OTHER WAY TO DO THIS, 5037 03:34:43,240 --> 03:34:45,480 IF I GIVE A RANDOM LIBRARY AND 5038 03:34:45,480 --> 03:34:47,400 TARGETED LIBRARY AND MIC THE TWO 5039 03:34:47,400 --> 03:34:49,720 AND IT WAS AGAINST A TARGET 5040 03:34:49,720 --> 03:34:53,240 WHERE AVERAGE ACTIVITY WAS ONE 5041 03:34:53,240 --> 03:34:54,680 NANOMOLAR I SHOULD HAVE BIMODAL 5042 03:34:54,680 --> 03:34:58,640 DISTRIBUTION IN MY SCREENING. SO 5043 03:34:58,640 --> 03:35:00,440 I BELABOR THIS BECAUSE THE 5044 03:35:00,440 --> 03:35:03,400 COMMON THING IS YOU DON'T FIND 5045 03:35:03,400 --> 03:35:04,800 ANYTHING SCREEN MUCH HIGHER 5046 03:35:04,800 --> 03:35:07,840 CONCENTRATION. THAT LEADS TO LOT 5047 03:35:07,840 --> 03:35:09,680 OF PROBLEMS IT SKEWS AND YOU 5048 03:35:09,680 --> 03:35:12,680 START TO EXPOSE ARTIFACTS OF 5049 03:35:12,680 --> 03:35:18,960 INSOLUBLE COMPOUNDS. SO AGAIN 5050 03:35:18,960 --> 03:35:21,920 BACK TO THIS ROBUSTNESS, THERE 5051 03:35:21,920 --> 03:35:22,880 IS DEBATE TRYING TO FIGURE HOW 5052 03:35:22,880 --> 03:35:28,600 TO DO THIS AND CALCULATE THIS. 5053 03:35:28,600 --> 03:35:29,640 SO THE FIRST THING YOU WANT TO 5054 03:35:29,640 --> 03:35:32,160 DO TO OPTIMIZE ASSAY IS REALLY 5055 03:35:32,160 --> 03:35:34,960 LOOK TO GET GAIN MIC RAKE HIGH. 5056 03:35:34,960 --> 03:35:39,760 SO THE BACKGROUND AND SIGNAL. 5057 03:35:39,760 --> 03:35:41,480 WHEN I STARTED MY CAREER 5058 03:35:41,480 --> 03:35:43,120 SCREENING GET BACKGROUND LOW 5059 03:35:43,120 --> 03:35:44,240 SIGNAL HIGH AND THAT IS A 5060 03:35:44,240 --> 03:35:45,520 PERFECT ASSAY. IT SEEMS 5061 03:35:45,520 --> 03:35:50,160 REASONABLE. BE AS I TALKED TO 5062 03:35:50,160 --> 03:35:50,960 STATISTICIANS THERE'S WAYS OF 5063 03:35:50,960 --> 03:35:56,800 DOING THIS. SO THE GROUP I HAD 5064 03:35:56,800 --> 03:35:58,200 AT DUPONT WE WERE ON ONE OF 5065 03:35:58,200 --> 03:36:01,800 THESE SOAS MEETINGS AND GOT ON 5066 03:36:01,800 --> 03:36:03,720 CRUISE BOAT AND WE WERE PRETTY 5067 03:36:03,720 --> 03:36:05,040 DRUNK. WE JUST STARTED TALKING 5068 03:36:05,040 --> 03:36:09,240 ABOUT THIS. AND MY SECOND IN 5069 03:36:09,240 --> 03:36:10,280 COMMAND SAID THERE SHOULD BE 5070 03:36:10,280 --> 03:36:11,840 REALLY SIMPLE PARAMETER, SHOULD 5071 03:36:11,840 --> 03:36:15,000 BE EASY TO REMEMBER. SO WE WERE 5072 03:36:15,000 --> 03:36:17,160 KICKING THE IDEA THIS IS A 5073 03:36:17,160 --> 03:36:19,640 COMMON THING. HERE IS AN ASSAY, 5074 03:36:19,640 --> 03:36:22,960 I WILL DEFINE WHAT IT IS IN A 5075 03:36:22,960 --> 03:36:25,440 SECOND BUT WE LOOK AT THIS 5076 03:36:25,440 --> 03:36:29,560 PARTICULAR ASSAY, IF THE SIGNAL 5077 03:36:29,560 --> 03:36:32,440 BACKGROUND IS HERE, THAT IS YOUR 5078 03:36:32,440 --> 03:36:35,240 BACKGROUND SIGNAL AND THEN YOUR 5079 03:36:35,240 --> 03:36:38,640 HIGH SIGNAL IS RIGHT HERE, THE 5080 03:36:38,640 --> 03:36:39,800 AVERAGE, THAT RATIO HERE FOR 5081 03:36:39,800 --> 03:36:40,960 THIS EXAMPLE IS TEN. SO THAT 5082 03:36:40,960 --> 03:36:43,720 SHOULD BE PRETTY GOOD ASSAY. 5083 03:36:43,720 --> 03:36:45,600 BUT LOOK AT THE SCATTER OF THE 5084 03:36:45,600 --> 03:36:47,280 DATA, ACTUAL COMPOUNDS AND 5085 03:36:47,280 --> 03:36:47,880 INTUITIVELY KNOW THAT IS NOT 5086 03:36:47,880 --> 03:36:51,360 GOOD. WHEREAS HERE YOU CAN SEE 5087 03:36:51,360 --> 03:36:53,920 THE HIGH SIGNAL CLUSTERS 5088 03:36:53,920 --> 03:36:55,200 TOGETHER, LOW SIGNAL CLUSTERING 5089 03:36:55,200 --> 03:37:01,360 TO AND WILL IS A FREE ZONE OF 5090 03:37:01,360 --> 03:37:04,080 BOUNDARY. SO THAT HAS SIGNAL TO 5091 03:37:04,080 --> 03:37:05,880 NOISE OF 12 AND SIGNAL TO 5092 03:37:05,880 --> 03:37:07,160 BACKGROUND OF ONLY FIVE BUT 5093 03:37:07,160 --> 03:37:09,440 CLEARLY LOOK AT THIS AND SAY 5094 03:37:09,440 --> 03:37:13,240 THAT IS A BETTER ASSAY. SO WE 5095 03:37:13,240 --> 03:37:16,680 WANTED TO FORMALIZE THIS. AND MY 5096 03:37:16,680 --> 03:37:19,480 GROUP TOLD ME THE SOLUTION AND I 5097 03:37:19,480 --> 03:37:20,520 THOUGHT THAT WAS OBVIOUS. I 5098 03:37:20,520 --> 03:37:22,120 THOUGHT IT WAS TRIVIAL AND I 5099 03:37:22,120 --> 03:37:26,120 SAID THAT'S STUPI STUPID. WHICHH 5100 03:37:26,120 --> 03:37:28,800 WE'LL USE IN THE GROUP BUT WHO 5101 03:37:28,800 --> 03:37:31,080 CARES. THE PERSON WHO WANTED TO 5102 03:37:31,080 --> 03:37:33,560 PUBLISH IT PUSHED AND PREDICTED 5103 03:37:33,560 --> 03:37:35,120 THERE WOULD BE MOST CITED PAPER 5104 03:37:35,120 --> 03:37:36,720 AND EVIDENTLY PROVIDE US 5105 03:37:36,720 --> 03:37:37,760 SOMETHING EASY FOR EVERYONE TO 5106 03:37:37,760 --> 03:37:42,440 USE SO THAT IS MY ADMISSION THAT 5107 03:37:42,440 --> 03:37:43,520 WASN'T THAT GOOD A MANAGER AT 5108 03:37:43,520 --> 03:37:48,560 THE TIME. SO I WANTED TO 5109 03:37:48,560 --> 03:37:50,040 FORMALLYZE THIS, THIS IT IS Z 5110 03:37:50,040 --> 03:37:51,160 PRIME YOU CAN LOOK IT UP. THE 5111 03:37:51,160 --> 03:37:54,480 WAY TO THINK ABOUT IT IS YOUR 5112 03:37:54,480 --> 03:37:56,080 BASICALLY THE HIGH SIGNAL AND 5113 03:37:56,080 --> 03:38:00,560 LOW SIGNAL CONTROLS, THE HIGH 5114 03:38:00,560 --> 03:38:01,960 CALL THIS LOW AND HIGH SIGNAL 5115 03:38:01,960 --> 03:38:03,440 CONTROLS, EACH HAVE A 5116 03:38:03,440 --> 03:38:05,640 DISTRIBUTION EACH HAS ITS 5117 03:38:05,640 --> 03:38:06,960 STANDARD DEVIATION AND SPREAD, 5118 03:38:06,960 --> 03:38:10,040 SO AS LONG AS EACH DISTRIBUTION 5119 03:38:10,040 --> 03:38:11,280 DOESN'T TOUCH EACH OTHER YOU 5120 03:38:11,280 --> 03:38:12,720 HAVE A GOOD ZONE OF SEPARATION 5121 03:38:12,720 --> 03:38:14,360 AND THE WAY TO THINK ABOUT IT IF 5122 03:38:14,360 --> 03:38:19,120 THE ZONE -- IF THIS VARIATION 5123 03:38:19,120 --> 03:38:20,920 WAS ZERO, STRAIGHT LINE, THIS 5124 03:38:20,920 --> 03:38:24,480 GOES TO ZERO AND Z PRIME WOULD 5125 03:38:24,480 --> 03:38:26,640 GO TO ONE. IT JUST MEANS IT IS 5126 03:38:26,640 --> 03:38:29,440 TWO LINES THEY DON'T EVER TOUCH. 5127 03:38:29,440 --> 03:38:30,560 THERE IS ONLY SEPARATION WILL BE 5128 03:38:30,560 --> 03:38:34,760 THE MAXIMUM IT CAN BE. DOESN'T 5129 03:38:34,760 --> 03:38:37,640 MATTER. THE OTHER PREMISE IS IF 5130 03:38:37,640 --> 03:38:40,560 YOU HAVE A -- THE ZONE OF 5131 03:38:40,560 --> 03:38:43,040 SEPARATION DISAPPEAR. WHAT THAT 5132 03:38:43,040 --> 03:38:45,360 MEANS IS THREE SIGMA LOW AND 5133 03:38:45,360 --> 03:38:48,400 THREE SIGMA HIGH BOTH TOUCH EACH 5134 03:38:48,400 --> 03:38:49,640 OTHER. AT TO POINT YOU STILL 5135 03:38:49,640 --> 03:38:55,960 HAVE ASSAY BUT IT IS BASICALLY 5136 03:38:55,960 --> 03:38:57,240 STILL ACCEPTED ASSAY BECAUSE YOU 5137 03:38:57,240 --> 03:39:01,240 ARE STILL -- WHERE THIS ZONE OF 5138 03:39:01,240 --> 03:39:02,880 SEPARATION APPEARS IT IS STILL 5139 03:39:02,880 --> 03:39:05,440 ONLY TOUCHING 1% VARIANCE. SO 5140 03:39:05,440 --> 03:39:07,160 YOU STILL HAVE THIS. BUT IF THE 5141 03:39:07,160 --> 03:39:09,880 TWO OVERLAP THEN THE -- IF THE 5142 03:39:09,880 --> 03:39:12,640 TWO ACTUALLY -- IF THIS ALL -- 5143 03:39:12,640 --> 03:39:15,560 IF THE SEPARATION OF MEANS IS 5144 03:39:15,560 --> 03:39:19,360 EQUAL TO THIS NUMERATOR TERM, 5145 03:39:19,360 --> 03:39:22,360 THIS COLLAPSES, SO IF THIS IS 5146 03:39:22,360 --> 03:39:25,680 ZERO HERE, THE TWO MEANS TOUCH, 5147 03:39:25,680 --> 03:39:27,480 NO MATTER WHAT THE DISTRIBUTION, 5148 03:39:27,480 --> 03:39:29,440 DISTRIBUTION IS OVERLAPPED. SO 5149 03:39:29,440 --> 03:39:32,440 IF -- THIS IS SEE REON THE 5150 03:39:32,440 --> 03:39:35,280 BOTTOM, THIS GOES DOWN TO Z, 5151 03:39:35,280 --> 03:39:36,960 ZERO. SO IT MEANS NO MATTER WHAT 5152 03:39:36,960 --> 03:39:40,240 YOU DO, THE SEPARATION OVERLAP. 5153 03:39:40,240 --> 03:39:42,120 SO THAT IS NOT A USABLE ASSAY. 5154 03:39:42,120 --> 03:39:46,120 BY DOING THIS YOU CAN CALCULATE 5155 03:39:46,120 --> 03:39:49,920 THAT Z -- IF Z FACTOR IS 1, 5156 03:39:49,920 --> 03:39:51,440 PERFECT ASSAY. YOU CAN REALLY 5157 03:39:51,440 --> 03:39:53,040 NEVER ACHIEVE THIS BECAUSE THAT 5158 03:39:53,040 --> 03:39:55,600 MEANS THERE'S NO NOISE IN EITHER 5159 03:39:55,600 --> 03:39:58,240 BAND BUT .5 TO 1 IS ACCEPTABLE 5160 03:39:58,240 --> 03:40:02,440 SCORE. AND IF IT IS BETWEEN 0 5161 03:40:02,440 --> 03:40:04,560 AND .5, YOU NEED TO DO 5162 03:40:04,560 --> 03:40:06,080 REPLICATES. YOU CAN SALVAGE 5163 03:40:06,080 --> 03:40:07,840 ASSAY BUT CAN'T RUN SINGLE KIT. 5164 03:40:07,840 --> 03:40:10,480 IF 0 OR LESS THAN 0 IT MEANS YOU 5165 03:40:10,480 --> 03:40:17,280 CANNOT DO AN ASSAY. IF IT IS 5166 03:40:17,280 --> 03:40:22,040 ZERO THE ACTUAL BANDS OVERLAP SO 5167 03:40:22,040 --> 03:40:23,360 MUCH IT IS IN OR OUT OF THE 5168 03:40:23,360 --> 03:40:24,960 SCREEN. SO THAT IS A YES, NO 5169 03:40:24,960 --> 03:40:29,720 ASSAY. THAT IS THE ORIGIN OF Z 5170 03:40:29,720 --> 03:40:36,040 FACTOR. HOW YOU USE IN REAL 5171 03:40:36,040 --> 03:40:38,120 SCREENS YOU MOST A HISTOGRAM OF 5172 03:40:38,120 --> 03:40:43,120 THE DATA. FREQUENCY PLOT OR PLOT 5173 03:40:43,120 --> 03:40:44,520 TYPICAL AND LOOK BASICALLY TO 5174 03:40:44,520 --> 03:40:45,920 SEE IF THERE IS A SEPARATION 5175 03:40:45,920 --> 03:40:48,320 BAND YOU CAN SEE IT THIS WAY, 5176 03:40:48,320 --> 03:40:53,200 SCATTER GRAM EMPHASIZES EACH 5177 03:40:53,200 --> 03:40:54,320 INDIVIDUAL DEFLECTION WHEREAS A 5178 03:40:54,320 --> 03:40:56,680 HISTOGRAM GIVES YOU DISTRIBUTION 5179 03:40:56,680 --> 03:40:58,200 OF THE AVERAGE SO YOU CAN SEE 5180 03:40:58,200 --> 03:41:01,520 HERE, THIS IS IDEALIZED. SO THE 5181 03:41:01,520 --> 03:41:03,240 CONTROL SHOULD BE TRULY GALSION 5182 03:41:03,240 --> 03:41:05,880 AS WELL AS HIGH AND LOW 5183 03:41:05,880 --> 03:41:07,280 CONTROLS. YOU LOOK TO SEE WHAT 5184 03:41:07,280 --> 03:41:09,000 HAPPENS WHEN YOU HAVE REAL 5185 03:41:09,000 --> 03:41:10,760 COMPOUNDS. IF YOU LOOK AT REAL 5186 03:41:10,760 --> 03:41:17,960 DATA HERE IS AN EXAMPLE OF ASSAY 5187 03:41:17,960 --> 03:41:19,720 PROTEIN PROTEIN INTERACTION, WE 5188 03:41:19,720 --> 03:41:20,880 CALCULATED THE PARAMETERS FOR 5189 03:41:20,880 --> 03:41:22,160 THE PIE WILL THE SCREEN, THIS IS 5190 03:41:22,160 --> 03:41:25,280 WHAT YOU DO, YOU CALCULATE THE 5191 03:41:25,280 --> 03:41:28,960 METRICS FOR PILOT SCREEN AND GET 5192 03:41:28,960 --> 03:41:31,240 VALUES Z FACTOR PRETTY GOOD, HAD 5193 03:41:31,240 --> 03:41:32,720 PRETTY GOOD SIGNAL BACKGROUND 5194 03:41:32,720 --> 03:41:36,400 THEN DO IT FOR THE FULL SCREEN 5195 03:41:36,400 --> 03:41:38,280 SO 7,000 COMPOUNDS, IN THIS 5196 03:41:38,280 --> 03:41:40,440 PARTICULAR CASE YOU CAN SEE THE 5197 03:41:40,440 --> 03:41:43,200 Z MAINTAINS ITSELF, THE POSITIVE 5198 03:41:43,200 --> 03:41:44,680 CONTROL SIGNALS WITHIN LINE SO 5199 03:41:44,680 --> 03:41:48,080 THIS IS GOOD. WE GOT REASONABLE 5200 03:41:48,080 --> 03:41:49,920 HIT RATES. WHEN YOU PLOT THE 5201 03:41:49,920 --> 03:41:51,880 ACTUAL DATA, PARAMETERS, YOU CAN 5202 03:41:51,880 --> 03:41:54,360 SEE HERE VERY CLEARLY, THE 5203 03:41:54,360 --> 03:41:59,400 CONTROLS ARE GALSION. THIS IS 5204 03:41:59,400 --> 03:42:02,160 BIASED BUT LOOK AT THE ENVELOPE 5205 03:42:02,160 --> 03:42:04,400 OF HITS, THE MAIN ENVELOPE IS 5206 03:42:04,400 --> 03:42:06,200 GALSION BUT YOU SEE THIS TAIL. 5207 03:42:06,200 --> 03:42:09,000 THIS IS GOOD. THIS PARTICULAR -- 5208 03:42:09,000 --> 03:42:10,840 THERE WAS ACTUALLY A WIDE RANGE 5209 03:42:10,840 --> 03:42:13,400 OF HITS. SO TYPICALLY WHAT YOU 5210 03:42:13,400 --> 03:42:16,000 SHOULD DO IS PLOT THIS AND THEN 5211 03:42:16,000 --> 03:42:26,520 PLOT BLOWUP OF INSET HERE. YOU 5212 03:42:30,280 --> 03:42:31,560 CAN SEE IT IN A SCATTER GRAM. 5213 03:42:31,560 --> 03:42:35,200 YOU CAN CALCULATE ON PLATE BY 5214 03:42:35,200 --> 03:42:36,600 PLATE BASIS SO THIS IS A GOOD 5215 03:42:36,600 --> 03:42:39,880 PORTION OF THE SCREEN, SO TEN 5216 03:42:39,880 --> 03:42:43,200 PLATE RUN AGO PLATE RUN ABOUT. 5217 03:42:43,200 --> 03:42:46,440 ABOUT 20,000 COMPOUNDS OR SO. 5218 03:42:46,440 --> 03:42:50,520 YOU CAN SEE FOR THIS ASSAY Z 5219 03:42:50,520 --> 03:42:52,800 FACTOR STAYED WITHIN SPEC, 5220 03:42:52,800 --> 03:42:56,320 ALWAYS GRADED .5 SIGNAL BACK 5221 03:42:56,320 --> 03:42:57,920 GROWN AND SIGNAL NOISE WORKS 5222 03:42:57,920 --> 03:43:00,120 CONSISTENT SO THIS GAVE US 5223 03:43:00,120 --> 03:43:01,640 CONFIDENCE THAT WE ARE PLATE BY 5224 03:43:01,640 --> 03:43:03,880 PLATE, RUN WAS GOOD SO WE CAN 5225 03:43:03,880 --> 03:43:05,240 AGGREGATE ALL THE DATA. THE 5226 03:43:05,240 --> 03:43:08,920 FINAL THING I WANT TO ASK YOU 5227 03:43:08,920 --> 03:43:11,720 WAS SO AGAIN THE REASON WHY WE 5228 03:43:11,720 --> 03:43:13,960 PICK THE ONE PERCENT, IT IS 5229 03:43:13,960 --> 03:43:15,120 THREE STANDARD DEVIATION AWAY 5230 03:43:15,120 --> 03:43:16,720 FROM THE POPULATION AVERAGE 5231 03:43:16,720 --> 03:43:21,200 WHICH IS RANDOM NOISE. SO IT IS 5232 03:43:21,200 --> 03:43:24,320 SIGNIFICANT THINGS. TYPICALLY IN 5233 03:43:24,320 --> 03:43:25,880 GROUPS YOU CAN SET A THRESHOLD 5234 03:43:25,880 --> 03:43:29,120 FOR CALLING A HIT STATISTICALLY 5235 03:43:29,120 --> 03:43:31,280 OR SOME DON'T CARE THEY JUST SAY 5236 03:43:31,280 --> 03:43:32,600 I WILL DO ARBITRARY 50% BECAUSE 5237 03:43:32,600 --> 03:43:36,080 IF I TESTED TEN MICROMOLAR AN 5238 03:43:36,080 --> 03:43:38,800 50% PERTURBATION IT IS A WEAK 5239 03:43:38,800 --> 03:43:40,720 MICROMOLAR HIT SO MOST COMPOUNDS 5240 03:43:40,720 --> 03:43:43,120 ARE PROBABLY MILLIMOLAR IC 5241 03:43:43,120 --> 03:43:45,400 50s. BUT IF YOU GO BY 5242 03:43:45,400 --> 03:43:47,680 STATISTICS IF YOU ARE HAVING A 5243 03:43:47,680 --> 03:43:49,160 THRESHOLD, IF YOU SET A 5244 03:43:49,160 --> 03:43:52,560 THRESHOLD AND THEN RETEST 5245 03:43:52,560 --> 03:43:54,040 COMPOUNDS, ANYTHING ABOVE THAT 5246 03:43:54,040 --> 03:43:55,520 THRESHOLD, IF YOU TEST THE 5247 03:43:55,520 --> 03:43:57,000 COMPOUND RIGHT AT THRESHOLD, 5248 03:43:57,000 --> 03:43:58,560 WHAT EVERY YOU CHOOSE FOR A 5249 03:43:58,560 --> 03:44:00,080 THRESHOLD, WHAT DO YOU THINK THE 5250 03:44:00,080 --> 03:44:01,040 CONFIRMATION RATE OF THAT 5251 03:44:01,040 --> 03:44:03,520 COMPOUND SHOULD BE? OR THOSE 5252 03:44:03,520 --> 03:44:04,800 COMPOUNDS? THEY ARE RIGHT AT 5253 03:44:04,800 --> 03:44:05,960 THRESHOLD. THE ANSWER IS HERE. 5254 03:44:05,960 --> 03:44:08,840 IT IS EXACTLY 50% BECAUSE YOU 5255 03:44:08,840 --> 03:44:10,960 RIGHT AT THRESHOLD SO IF YOU 5256 03:44:10,960 --> 03:44:13,320 RETEST IT NO MATTER HOW PRECISE 5257 03:44:13,320 --> 03:44:14,840 HALF THE TIME ABOVE, HALF THE 5258 03:44:14,840 --> 03:44:18,760 TIME BELOW. HOWEVER IF YOU HAVE 5259 03:44:18,760 --> 03:44:21,880 A COMPOUND THAT IS SAY FIVE 5260 03:44:21,880 --> 03:44:24,640 STANDARD DEVIATION FROM ORIGINAL 5261 03:44:24,640 --> 03:44:30,960 SIGMA, AND YOU RETEST THAT 5262 03:44:30,960 --> 03:44:32,680 COMPOUND, 500 REPLICATES AND 5263 03:44:32,680 --> 03:44:35,680 RETEST IT, IT SHOULD BE NORMAL 5264 03:44:35,680 --> 03:44:36,960 DISTRIBUTION. IT IS VARIATION 5265 03:44:36,960 --> 03:44:38,440 WILL BE DEPENDENT UPON THE 5266 03:44:38,440 --> 03:44:44,280 RANDOM ERROR OF YOUR SAY SAY. 5267 03:44:44,280 --> 03:44:45,560 THERE IS A WAY TO HAVE 5268 03:44:45,560 --> 03:44:47,160 CALCULATING IF YOU HAVE A TABLE 5269 03:44:47,160 --> 03:44:49,800 OF ALL YOUR HITS AND THERE ARE 5270 03:44:49,800 --> 03:44:52,400 PERCENT INHIBITION, YOU CAN 5271 03:44:52,400 --> 03:44:55,800 CALCULATE THE INHIBITION 5272 03:44:55,800 --> 03:44:57,520 CONFIRMATION RATE EXPECTED. 5273 03:44:57,520 --> 03:44:59,520 PRACTICALLY YOU WILL NEVER HIT 5274 03:44:59,520 --> 03:45:00,840 50% COPYMATION BECAUSE THAT 5275 03:45:00,840 --> 03:45:01,920 MEANS ALREADY HITS THRESHOLD. 5276 03:45:01,920 --> 03:45:08,200 WHAT IS REASONABLE FOR THRESHOLD 5277 03:45:08,200 --> 03:45:10,120 IS ABOUT 30% OVERALL 5278 03:45:10,120 --> 03:45:11,720 CONFIRMATION IS WHAT YOU CAN 5279 03:45:11,720 --> 03:45:14,360 GET. YOU CAN GET FEWER POTENT 5280 03:45:14,360 --> 03:45:21,600 HITS, MOST THRESHOLD. WHEN I DO 5281 03:45:21,600 --> 03:45:23,840 A SCREEN, IF I GET CONFIRMATION 5282 03:45:23,840 --> 03:45:29,960 RATES BELOW 30% I WORRY THAT IS 5283 03:45:29,960 --> 03:45:31,640 WHEN YOU WANT REPLY CASE AND GO 5284 03:45:31,640 --> 03:45:39,960 BACK TO CHECK CONDITIONS. LAST 5285 03:45:39,960 --> 03:45:41,280 IS FAST POSITIVE FALSE NEGATIVE. 5286 03:45:41,280 --> 03:45:44,680 IF YOU SET A THRESHOLD TOO HIGH, 5287 03:45:44,680 --> 03:45:46,880 TOO STRINGENTLY, THERE IS A 5288 03:45:46,880 --> 03:45:47,720 CHANCE YOU YOU WILL MISS 5289 03:45:47,720 --> 03:45:50,960 IMPORTANT COMPOUNDS. THE WAY YOU 5290 03:45:50,960 --> 03:45:54,160 DO THAT IS REDUCE ITS 5291 03:45:54,160 --> 03:45:56,960 STRINGENCY, LET THE THRESHOLD BE 5292 03:45:56,960 --> 03:45:59,360 LOWER SO MORE HITS ARE INCLUDED 5293 03:45:59,360 --> 03:46:00,880 IT DOES INCREASE YOUR FALSE 5294 03:46:00,880 --> 03:46:04,360 POSITIVE RATES BUT I THINK IF 5295 03:46:04,360 --> 03:46:07,960 YOU MISS A COMPOUND CLASS, THE 5296 03:46:07,960 --> 03:46:10,200 FALSE NEGATIVE IS MORE DAMAGING 5297 03:46:10,200 --> 03:46:11,520 THAN TOO MANY FALSE POSITIVES SO 5298 03:46:11,520 --> 03:46:13,600 YOU CAN FIND THAT YOU CAN DO A 5299 03:46:13,600 --> 03:46:15,840 SCREEN AND TEST HIT RATE COUPLE 5300 03:46:15,840 --> 03:46:17,240 OF DIFFERENT THRESHOLDS AN CHECK 5301 03:46:17,240 --> 03:46:20,480 YOUR CONFIRMATION AND SCAFFOLDS 5302 03:46:20,480 --> 03:46:21,520 YOU CAN COME UP WITH AND PLAY 5303 03:46:21,520 --> 03:46:31,080 WITH THAT. JUST TO SUMMARIZE ONE 5304 03:46:31,080 --> 03:46:32,360 THING I HEAR IS LOT OF PLACES 5305 03:46:32,360 --> 03:46:33,880 THEY HAVE A HIGH THROUGH PUT 5306 03:46:33,880 --> 03:46:37,320 GROUP FOCUSED ON HIGH PRODUCTION 5307 03:46:37,320 --> 03:46:38,400 AND THEY HAVE A DOSE RESPONSE 5308 03:46:38,400 --> 03:46:42,880 GROUP. BUT YOU SHOULD USE 5309 03:46:42,880 --> 03:46:46,320 PRIMARY ASSAY AS YOUR IC 50 5310 03:46:46,320 --> 03:46:47,480 GENERATING ASSAY. WHICH IS WHAT 5311 03:46:47,480 --> 03:46:53,160 WE DO. YOU CAN SEE IF YOU HAVE A 5312 03:46:53,160 --> 03:46:55,960 1536 WELL PLATE YOU CAN SUPPORT 5313 03:46:55,960 --> 03:46:59,000 QUITE A NUMBER OF DOSE RESPONSE 5314 03:46:59,000 --> 03:47:02,800 ASSAYS 10-POINT DOSE RESPONSE, 5315 03:47:02,800 --> 03:47:05,200 129 IN TYPICAL 36 WELL PLATE SO 5316 03:47:05,200 --> 03:47:11,240 NEVER LIMITING THE CYCLE TIME 5317 03:47:11,240 --> 03:47:12,680 WHENEVER WE DESIGN ASSAYS WE 5318 03:47:12,680 --> 03:47:14,440 DESIGN BOTH PRIMARY AND COUNTER 5319 03:47:14,440 --> 03:47:16,240 ASSAY IN THESE HIGH THROUGH PUT 5320 03:47:16,240 --> 03:47:17,480 FORMATS SO WE CAN SUPPORT THIS 5321 03:47:17,480 --> 03:47:25,840 THING. AND THEN WE FOCUS ON DO 5322 03:47:25,840 --> 03:47:27,400 THE DIFFERENT BIOLOGICAL ASSAYS 5323 03:47:27,400 --> 03:47:31,400 LOWER THROUGH PUT BUT TRIAGE 5324 03:47:31,400 --> 03:47:32,480 DOWN TO REAL SCAFFOLDS THAT ARE 5325 03:47:32,480 --> 03:47:37,760 SHOWING SAR. SO WHEN YOU CAN DO 5326 03:47:37,760 --> 03:47:40,560 THAT ON -- IN THE NCATS HAS DONE 5327 03:47:40,560 --> 03:47:45,000 THIS, BECAUSE THEY SCREEN BUY 5328 03:47:45,000 --> 03:47:45,800 LIBRARIES VERTICALLY THEY HAVE 5329 03:47:45,800 --> 03:47:46,800 FULL DOSE RESPONSE CURVE AND 5330 03:47:46,800 --> 03:47:48,480 TEACHING IS YOU CAN'T LOOK AT 5331 03:47:48,480 --> 03:47:49,760 THAT MANY CURVES YOU HAVE TO 5332 03:47:49,760 --> 03:47:53,240 CATEGORIZE THE CURVES TURNS OUT 5333 03:47:53,240 --> 03:47:54,440 THAT YOU ARE CATEGORIZE WITH 5334 03:47:54,440 --> 03:47:56,320 DESCRIPTORS THAT YOU LOOK UP IN 5335 03:47:56,320 --> 03:47:59,360 THE NCATS ASSAY GUIDANCE MANUAL 5336 03:47:59,360 --> 03:48:06,040 ON HGS IC 50 BASED HCS BUT THIS 5337 03:48:06,040 --> 03:48:07,360 IS IMPORTANT FOR COLLEAGUES 5338 03:48:07,360 --> 03:48:09,280 BECAUSE YOU CAN HAVE COMPOUNDS 5339 03:48:09,280 --> 03:48:12,680 GIVE YOU FULL NORMAL RESPONSE 5340 03:48:12,680 --> 03:48:14,560 WITH TWO LOW AND HIGH, ALSO ONES 5341 03:48:14,560 --> 03:48:18,520 THAT REACH LOWER ASIMTOTE OR 5342 03:48:18,520 --> 03:48:21,320 DON'T BOUND OR HAVE MORE THAN 5343 03:48:21,320 --> 03:48:24,320 ONE MAXIMUM. . BUT THEY HAVE 5344 03:48:24,320 --> 03:48:30,200 FORMALIZED ALL THIS. SO FINAL 5345 03:48:30,200 --> 03:48:33,320 COMMENTS, JUST THAT HGS RELIES 5346 03:48:33,320 --> 03:48:36,040 ON STATISTICALLY BASED ROLES AND 5347 03:48:36,040 --> 03:48:37,840 BIGGEST IS MOST COMPOUNDS ARE 5348 03:48:37,840 --> 03:48:38,960 INACTIVE SO THEY ACT LIKE 5349 03:48:38,960 --> 03:48:40,240 BACKGROUND NOISE. YOU CAN TREAT 5350 03:48:40,240 --> 03:48:42,840 THAT NORMALLY. THE OTHER THING, 5351 03:48:42,840 --> 03:48:44,000 YOU CAN SPEND TIME TRYING TO 5352 03:48:44,000 --> 03:48:46,560 MAKE THE ASSAY PERFECT BUT YOU 5353 03:48:46,560 --> 03:48:47,600 ONLY NEED TO MAKE IT GOOD ENOUGH 5354 03:48:47,600 --> 03:48:49,600 AND ROBUST ENOUGH TO BE 5355 03:48:49,600 --> 03:48:52,080 CONFIDENT THAT A SINGLE 5356 03:48:52,080 --> 03:48:53,200 CONCENTRATION ATTEMPT FINDING 5357 03:48:53,200 --> 03:48:54,440 THAT COMPOUND YOU HAVE A GOOD 5358 03:48:54,440 --> 03:48:57,240 CHANCE OF FINDING IT. THE OTHER 5359 03:48:57,240 --> 03:49:00,240 THING THAT PROTECTS ALL OF US IN 5360 03:49:00,240 --> 03:49:02,800 REDUCE SCREENING IS IT IS VERY 5361 03:49:02,800 --> 03:49:03,920 RARE YOU SCREEN CHEMICAL 5362 03:49:03,920 --> 03:49:04,880 COLLECTION IS IS THAT IS 5363 03:49:04,880 --> 03:49:06,560 COMPOSED OF SINGLETONS THAT 5364 03:49:06,560 --> 03:49:08,520 DON'T HAVE RELATED ANALOGS IN 5365 03:49:08,520 --> 03:49:11,360 THAT. GOOD LIBRARIES ARE BASED 5366 03:49:11,360 --> 03:49:13,360 UPON ISLANDS DIVERSITY AND 5367 03:49:13,360 --> 03:49:15,680 OTHERS SAR. AND YOU CAN MITIGATE 5368 03:49:15,680 --> 03:49:18,480 THE RISK OF FALSE POSITIVES, 5369 03:49:18,480 --> 03:49:23,280 FALSE POSITIVE BY SEEING HIGHER 5370 03:49:23,280 --> 03:49:25,160 THRESHOLD BUT YOU HAVE TO THINK 5371 03:49:25,160 --> 03:49:25,880 FALSE NEGATIVES YOU MIGHT HAVE. 5372 03:49:25,880 --> 03:49:27,640 FINALLY, WHEN WE ARE DOING 5373 03:49:27,640 --> 03:49:29,240 CONFIRMATION WE DO REPLICATES 5374 03:49:29,240 --> 03:49:33,280 BECAUSE THAT GIVES YOU A CHANCE 5375 03:49:33,280 --> 03:49:35,600 TO VERIFY WHAT YOU FOUND WAS AN 5376 03:49:35,600 --> 03:49:38,920 EFFECT WITHIN THAT POPULATION. 5377 03:49:38,920 --> 03:49:41,880 LAST THING, IF YOU FOLLOW THESE 5378 03:49:41,880 --> 03:49:42,680 RULES YOU SHOULD ORBED BE 5379 03:49:42,680 --> 03:49:46,560 CONFIDENT YOUR ASSAY FOUND 5380 03:49:46,560 --> 03:49:47,840 CHEMICAL COMPOUNDS YOU SHOULDN'T 5381 03:49:47,840 --> 03:49:49,560 HAVE BEEN ABLE TO FIND, IT IS 5382 03:49:49,560 --> 03:49:52,840 JUST AS IMPORTANT TO TELL 5383 03:49:52,840 --> 03:49:54,480 MANAGEMENT AND PRESERVE 5384 03:49:54,480 --> 03:49:58,240 RESOURCES THAT YOU SCREEN 5385 03:49:58,240 --> 03:49:59,160 CORRECTLY WHAT IS PROBLEM IS NOT 5386 03:49:59,160 --> 03:50:00,640 THE ASSAY, THE COLLECTION OF 5387 03:50:00,640 --> 03:50:03,000 COMPOUNDS YOU TEST. BECAUSE LIKE 5388 03:50:03,000 --> 03:50:04,880 FISHING IF YOU ARE IN A LAKE 5389 03:50:04,880 --> 03:50:06,480 THAT DOESN'T HAVE FISH, DOESN'T 5390 03:50:06,480 --> 03:50:09,760 MATTER HOW WELL YOU FISH. 5391 03:50:09,760 --> 03:50:11,720 ANYWAY, THAT IS MY LITTLE 5392 03:50:11,720 --> 03:50:12,040 TUTORIAL. 5393 03:50:12,040 --> 03:50:19,560 [APPLAUSE] 5394 03:50:19,560 --> 03:50:21,960 >>THANK YOU SO MUCH, TC FOR A 5395 03:50:21,960 --> 03:50:23,320 WONDERFUL TALK. QUESTIONS. 5396 03:50:23,320 --> 03:50:24,920 >>CAN YOU PLEASE COME CLOSE TO 5397 03:50:24,920 --> 03:50:28,320 THE MICROPHONE? I HAVE BEEN TOLD 5398 03:50:28,320 --> 03:50:28,600 TO SAY THAT. 5399 03:50:28,600 --> 03:50:31,480 >>THANK YOU VERY MUCH FOR THAT 5400 03:50:31,480 --> 03:50:33,240 WONDERFUL PRESENTATION. YOU 5401 03:50:33,240 --> 03:50:38,440 MENTIONED THAT FOR THE Z 5402 03:50:38,440 --> 03:50:39,680 STATISTICS WORK THERE RECOGNIZE 5403 03:50:39,680 --> 03:50:41,600 TO BE TEXT ON YOUR SLIDE THAT 5404 03:50:41,600 --> 03:50:45,200 NEEDS TO BE LARGE SAMPLE. I KNOW 5405 03:50:45,200 --> 03:50:46,600 THIS MIGHT BE NOT POSSIBLE TO 5406 03:50:46,600 --> 03:50:48,160 ANSWER BUT FROM YOUR EXPERIENCE 5407 03:50:48,160 --> 03:50:50,440 DO YOU HAVE A RULE OF THUMB FOR 5408 03:50:50,440 --> 03:50:52,680 HOW LARGE A SAMPLE YOU NEED TO 5409 03:50:52,680 --> 03:50:54,560 GET RELIABLE NUMBER? 5410 03:50:54,560 --> 03:50:56,600 >>DEFAULT TO THE SAMPLE MEAN, 5411 03:50:56,600 --> 03:50:59,600 SO IF YOU ARE GREATER THAN 30 5412 03:50:59,600 --> 03:51:02,680 YOU REALLY FIND. WHEN WE WERE IN 5413 03:51:02,680 --> 03:51:05,640 96 WELL PLACE, YOU ARE 5414 03:51:05,640 --> 03:51:06,920 CONSTRAINED. LOOKING AT ONE 5415 03:51:06,920 --> 03:51:09,720 PLATE YOU ARE CON VAINED TO 5416 03:51:09,720 --> 03:51:11,240 EIGHT BY 12 SO MAYBE FOUR AND 5417 03:51:11,240 --> 03:51:16,320 FOUR, THAT'S LOW BUT THEN WE 5418 03:51:16,320 --> 03:51:19,000 WOULD WHEN SET UP ASSAY TAKE 9 # 5419 03:51:19,000 --> 03:51:22,400 RUN HALF HIGH, HALF LOW. YOU 5420 03:51:22,400 --> 03:51:23,800 MAKE THE AIONAL WHEN YOU PUT 5421 03:51:23,800 --> 03:51:28,320 REAL COMPOUNDS ON YOU INCLUDE 5422 03:51:28,320 --> 03:51:33,480 SOME THOSE REPLICATES AS WELL. 5423 03:51:33,480 --> 03:51:35,200 WHAT YOU DO IS YOU DO PLATE BY 5424 03:51:35,200 --> 03:51:36,320 PLATE THEN AT THE END YOU CAN 5425 03:51:36,320 --> 03:51:38,360 LOOK AT ALL YOUR CONTROLS AND 5426 03:51:38,360 --> 03:51:39,520 PULL THEM TOGETHER AND THAT 5427 03:51:39,520 --> 03:51:41,040 GIVES YOU OVERALL STATISTIC AS 5428 03:51:41,040 --> 03:51:47,080 WELL. IF YOU RUNNING ANIST 5429 03:51:47,080 --> 03:51:49,160 ASSAYS A FOR FAN GP YOU HAVE TO 5430 03:51:49,160 --> 03:51:50,520 REFERENCE HEIGHTENED CONTROL, SO 5431 03:51:50,520 --> 03:51:53,520 TWO WAYS TO DO THAT, USE A Z 5432 03:51:53,520 --> 03:51:55,200 SCORE WHICH MEANS HERE IS MY 5433 03:51:55,200 --> 03:52:00,680 FLOOR OF THE ASSAY, AND POP UP 5434 03:52:00,680 --> 03:52:02,680 ABOVE THAT. BY EXPERIENCE YOU 5435 03:52:02,680 --> 03:52:05,400 CAN HAVE TIGHT THINGS AND THINGS 5436 03:52:05,400 --> 03:52:10,440 THAT RISE TEN PERCENT ABOVE 5437 03:52:10,440 --> 03:52:13,600 BASELINE BUT NEVER GIVE A HIT 5438 03:52:13,600 --> 03:52:15,200 BUT YOU CAN STATISTICALLY DO 5439 03:52:15,200 --> 03:52:19,600 THAT. THE OTHER WAY FAKE IS LOW 5440 03:52:19,600 --> 03:52:20,640 CELL COUNT FOR HIGH CELL COUNT 5441 03:52:20,640 --> 03:52:21,760 RECEPTOR AND FORCE THE SYSTEM 5442 03:52:21,760 --> 03:52:23,600 BUT YOU DONE KNOW UNLESS YOU 5443 03:52:23,600 --> 03:52:29,840 HAVE A REFERENCE ACTNIST. 5444 03:52:29,840 --> 03:52:32,000 >>I HAD A QUESTION GIVEN YOUR 5445 03:52:32,000 --> 03:52:35,120 COMMENT THAT MOST COMPOUNDS ARE 5446 03:52:35,120 --> 03:52:36,240 INACTIVE, 99% WHATEVER THE 5447 03:52:36,240 --> 03:52:38,480 NUMBER MIGHT BE, HOW MUCH OF THE 5448 03:52:38,480 --> 03:52:39,920 ASSAY OPTIMIZATION CAN BE 5449 03:52:39,920 --> 03:52:41,840 DEFERRED TO THE ACTUAL SCREEN? 5450 03:52:41,840 --> 03:52:44,120 LET ME ASK SPECIFIC QUESTION, 5451 03:52:44,120 --> 03:52:46,600 SPECIFIC EXAMPLE, SAY YOU TRY TO 5452 03:52:46,600 --> 03:52:47,280 ASSESS PLATE TO PLATE 5453 03:52:47,280 --> 03:52:49,600 VARIABILITY. IN THE ACTUAL 5454 03:52:49,600 --> 03:52:51,640 SCREEN YOU SHOULD SEE THAT 5455 03:52:51,640 --> 03:52:53,080 ANYWAYS AND BECAUSE YOU HAVE GOT 5456 03:52:53,080 --> 03:52:54,120 ALL THESE DIFFERENT PLACE 5457 03:52:54,120 --> 03:52:55,840 RUNNING SO DOES THAT NEED TO BE 5458 03:52:55,840 --> 03:52:56,920 DONE DURING OPTIMIZATION BECAUSE 5459 03:52:56,920 --> 03:52:58,520 YOU WILL SEE IT ANY WAYS IN THE 5460 03:52:58,520 --> 03:53:02,680 ACTUAL SCREEN? 5461 03:53:02,680 --> 03:53:05,200 >>THE OPTIMIZATION WITH SMALL 5462 03:53:05,200 --> 03:53:06,360 NUMBER OF REPRESENTED PLACE WILL 5463 03:53:06,360 --> 03:53:09,200 GIVE YOU THE LOWEST SOMETHING 5464 03:53:09,200 --> 03:53:11,920 CAN BE WITHOUT INTERFERENCE. YOU 5465 03:53:11,920 --> 03:53:15,200 HEARD WHILE COMPOUNDS -- 5466 03:53:15,200 --> 03:53:18,240 ASSUMPTION HYPOTHESIS MOST 5467 03:53:18,240 --> 03:53:21,080 COMPOUNDS INACTIVE, THAT IS 5468 03:53:21,080 --> 03:53:23,240 BASED UPON ACTIVITY OF TARGET 5469 03:53:23,240 --> 03:53:24,480 INVOLVED IN BUT AS YOU HEARD 5470 03:53:24,480 --> 03:53:26,880 HERE THERE IS COMPOUND 5471 03:53:26,880 --> 03:53:28,120 INTERFERENCE THAT HAS NOTHING TO 5472 03:53:28,120 --> 03:53:35,280 DO WITH BIOLOG BIOLOGY. INTERFEG 5473 03:53:35,280 --> 03:53:38,240 WITH THE SYSTEM, NON-BIOLOGICAL 5474 03:53:38,240 --> 03:53:41,760 BUT REPRODUCIBLE. TO DEFER, BACK 5475 03:53:41,760 --> 03:53:44,600 -- YOU TAKE ALL YOUR COMPOUNDS 5476 03:53:44,600 --> 03:53:46,200 LOOK AT NUMBERS OF HEAT MAPS AND 5477 03:53:46,200 --> 03:53:52,720 LOOK AT THEM. IF THERE IS HIGH 5478 03:53:52,720 --> 03:53:53,760 COMPOUNDS ON THIS SIDE OR 5479 03:53:53,760 --> 03:53:54,800 STRIPING YOU WILL SEE THOSE 5480 03:53:54,800 --> 03:53:56,400 PATTERNS. IF U YOU SEE LIKE -- 5481 03:53:56,400 --> 03:53:58,200 IF YOU HAVE IMAGE BASED ASSAY, 5482 03:53:58,200 --> 03:54:02,280 CENTER OF THE PLATE JUST BLOOMS, 5483 03:54:02,280 --> 03:54:07,480 YOU WILL BE WARNED. IF YOU ARE 5484 03:54:07,480 --> 03:54:09,280 AT A VALUABLE TARGET WHERE 5485 03:54:09,280 --> 03:54:11,360 EVERYTHING IS EXPENSIVE, YOU CAN 5486 03:54:11,360 --> 03:54:15,000 TAKE SHORT CUTS LIKE THAT. 5487 03:54:15,000 --> 03:54:18,880 >>ANY MORE QUESTIONS FOR TC? I 5488 03:54:18,880 --> 03:54:20,240 DON'T SEE MORE QUESTIONS THANK 5489 03:54:20,240 --> 03:54:21,480 YOU SO MUCH, TC. APPRECIATE YOUR 5490 03:54:21,480 --> 03:54:21,840 TIME. 5491 03:54:21,840 --> 03:54:32,040 [APPLAUSE] 5492 03:54:40,600 --> 03:54:46,440 OUR NEXT SPEAKER IS DR. 5493 03:54:46,440 --> 03:54:48,000 VISWANATH DEVANARAYAN, EXECUTIVE 5494 03:54:48,000 --> 03:54:49,760 DIRECTOR AND HEAD OF DATA 5495 03:54:49,760 --> 03:54:53,160 SCIENCE AT ASCI INC., ALSO 5496 03:54:53,160 --> 03:54:54,200 ADJUNCT PROFESSOR UNIVERSITY OF 5497 03:54:54,200 --> 03:55:00,400 ILLINOIS CHICAGO. HE HAS MORE 5498 03:55:00,400 --> 03:55:01,960 THAN 25 YEARS RESEARCH 5499 03:55:01,960 --> 03:55:08,560 EXPERIENCE OF GSK ELY LILLY, 5500 03:55:08,560 --> 03:55:10,920 MERC. DR. DEVANARAYAN IS A LONG 5501 03:55:10,920 --> 03:55:13,040 STANDING EDITORIAL BOARD MEMBER, 5502 03:55:13,040 --> 03:55:14,120 WE ARE EXCITED TO HAVE HIM HERE 5503 03:55:14,120 --> 03:55:17,560 TODAY. HE WILL BE SPEAKING 5504 03:55:17,560 --> 03:55:19,760 ABOUT REPRODUCIBILITY ASSESSMENT 5505 03:55:19,760 --> 03:55:21,920 OF IN VITRO SCREENING RESULTS. 5506 03:55:21,920 --> 03:55:32,440 FLOOR IS YOURS, DEVANARAYAN. 5507 03:55:44,560 --> 03:55:47,600 MANY TIMES SELENIAN ABIGALE PUT 5508 03:55:47,600 --> 03:55:48,600 IN HARD WORK ORGANIZING THE 5509 03:55:48,600 --> 03:55:50,960 WORKSHOP. BEEN DOING THIS A LONG 5510 03:55:50,960 --> 03:55:53,880 TIME AND EACH TIME A DIFFERENT 5511 03:55:53,880 --> 03:55:55,680 PLACE, THIS PLACE IS FANTASTIC, 5512 03:55:55,680 --> 03:55:57,240 VERY NICELY SPREAD OUT AND 5513 03:55:57,240 --> 03:55:58,200 COMFORTABLE AND THANK YOU ALL 5514 03:55:58,200 --> 03:56:04,880 FOR MAKING IT. TO THIS WORKSHOP 5515 03:56:04,880 --> 03:56:06,640 THIS IS MY FIRST TALK IN PERSON 5516 03:56:06,640 --> 03:56:08,280 SINCE THE PANDEMIC STARTED. IT'S 5517 03:56:08,280 --> 03:56:12,640 BEEN SO LONG, SO LONG BEFORE I 5518 03:56:12,640 --> 03:56:15,360 USED TO GIVE MULTIPLE TALKS 5519 03:56:15,360 --> 03:56:17,400 EVERY YEAR. IT'S BEEN TWO YEARS, 5520 03:56:17,400 --> 03:56:19,240 TWO OR THREE YEARS NOW AND BEEN 5521 03:56:19,240 --> 03:56:25,320 THE FIRST TIME IN PUBLIC. SO 5522 03:56:25,320 --> 03:56:26,760 REAL EXCITED, THAT WE ARE 5523 03:56:26,760 --> 03:56:27,880 GETTING CLOSER TO THE NEW 5524 03:56:27,880 --> 03:56:33,800 NORMAL. YOU HEARD ABOUT Z PRIME 5525 03:56:33,800 --> 03:56:35,400 FACTOR AND ALL THE IMPORTANT 5526 03:56:35,400 --> 03:56:37,720 STATISTICAL METRICS FROM TC. 5527 03:56:37,720 --> 03:56:39,240 FOR THE LAST 45 MINUTES. I'M 5528 03:56:39,240 --> 03:56:44,160 GOING TO EXTEND THAT FURTHER TO 5529 03:56:44,160 --> 03:56:48,280 THE SCREENING ASSAYS WHERE WE 5530 03:56:48,280 --> 03:56:50,040 WORRY ABOUT THE VARIABILITY OF 5531 03:56:50,040 --> 03:56:51,320 POTENCY RESULTS FROM VARIOUS 5532 03:56:51,320 --> 03:56:55,320 COMPOUNDS THAT ARE TESTED. EC 50 5533 03:56:55,320 --> 03:56:56,680 IC 50, K, AND SO FORTH. SO THERE 5534 03:56:56,680 --> 03:57:00,200 IS ANOTHER METRIC LIKE Z PRIME 5535 03:57:00,200 --> 03:57:07,920 NOT AS POPULAR. BUT RELEVANT FOR 5536 03:57:07,920 --> 03:57:09,400 COMPARING POTENCY RESULTS IN 5537 03:57:09,400 --> 03:57:13,280 COMPOUNDS. SO WHAT WE ARE DOING 5538 03:57:13,280 --> 03:57:14,680 IS DIFFERENTIATING COMPOUNDS IN 5539 03:57:14,680 --> 03:57:16,840 TERMS OF METRICS AND RANK 5540 03:57:16,840 --> 03:57:18,080 ORDERING THEM AND DECIDING WHAT 5541 03:57:18,080 --> 03:57:22,760 TO MOVE FORWARD. LOOK AT 5542 03:57:22,760 --> 03:57:24,120 STANDARD ERROR ASSOCIATED WITH 5543 03:57:24,120 --> 03:57:27,800 KI OR IC 50 OR DC 50 AND MAKE 5544 03:57:27,800 --> 03:57:28,800 APPROXIMATE JUDGMENT BUT IS 5545 03:57:28,800 --> 03:57:33,760 THERE A Z PRIME THAT WECAL QUEUE 5546 03:57:33,760 --> 03:57:36,440 RATE FOR SCREENING DAT DATA. AFR 5547 03:57:36,440 --> 03:57:37,480 CALCULATING THAT WE CAN FEEL 5548 03:57:37,480 --> 03:57:38,720 CONFIDENT THIS SCREENING DATA 5549 03:57:38,720 --> 03:57:41,880 ASSAY WILL BE RELIABLE FOR 5550 03:57:41,880 --> 03:57:43,840 GENERATING THOSE EC 50 IC 50 5551 03:57:43,840 --> 03:57:45,400 VALUES FOR THOSE COMPOUNDS. SO 5552 03:57:45,400 --> 03:57:48,040 THERE IS A METRIC LIKE THAT, IT 5553 03:57:48,040 --> 03:57:51,880 IS HARDER TO CALCULATE, THAN Z 5554 03:57:51,880 --> 03:57:54,440 PRIME, BUT WE GO TONA IN SOME 5555 03:57:54,440 --> 03:57:57,760 DETAIL NEXT 40 MINUTES. THIS IS 5556 03:57:57,760 --> 03:58:01,000 COVERED IN COUPLE OF CHAPTERS IN 5557 03:58:01,000 --> 03:58:04,800 THE AGM. THERE ARE SOME 5558 03:58:04,800 --> 03:58:06,280 MATERIAL IN THE SLIDES NOT 5559 03:58:06,280 --> 03:58:07,640 COVERED BUT IF YOU HAVE ANY 5560 03:58:07,640 --> 03:58:08,560 QUESTIONS FEEL FREE TO REACH OUT 5561 03:58:08,560 --> 03:58:13,360 LATER. HOW MANY HEARD Z PRIME? 5562 03:58:13,360 --> 03:58:16,680 MOST OF YOU HAVE. HOW MANY HEARD 5563 03:58:16,680 --> 03:58:20,240 ABOUT MSR? MEAN SIGNIFICANT 5564 03:58:20,240 --> 03:58:24,840 RATIO? ONE, TWO, THREE, FOUR. 5565 03:58:24,840 --> 03:58:26,760 AND I HAVE BEEN PRESENTING THIS 5566 03:58:26,760 --> 03:58:30,440 SINCE 2005 AT SBS AND SOAS AND 5567 03:58:30,440 --> 03:58:34,360 THESE WORKSHOPS AND AGAIN THIS 5568 03:58:34,360 --> 03:58:36,760 IS NOT AS EASY TO CALCULATE A Z 5569 03:58:36,760 --> 03:58:39,760 PRIME AND PROBABLY WHY NOT 5570 03:58:39,760 --> 03:58:41,640 CAUGHT ON YET BUT AS BACKGROUND 5571 03:58:41,640 --> 03:58:44,960 I USED TO WORK AT ELY LILLY, 5572 03:58:44,960 --> 03:58:45,720 MERCK SO FORTH ALL THESE PLACE 5573 03:58:45,720 --> 03:58:47,480 WHERE IS I WORKED WHEN THIS 5574 03:58:47,480 --> 03:58:49,200 CONCEPT WAS INTRODUCED MSR 5575 03:58:49,200 --> 03:58:52,280 CONCEPT WAS INTRODUCED, THERE 5576 03:58:52,280 --> 03:58:53,840 WAS ALWAYS SOME DELAY ADOPTING 5577 03:58:53,840 --> 03:58:55,720 THAT BUT ONCE PEOPLE GOT USED TO 5578 03:58:55,720 --> 03:58:59,400 IT, THEY JUST LOVED IT AND NOW 5579 03:58:59,400 --> 03:59:01,760 COUPLE OF YEARS BECAME REQUIRE 5580 03:59:01,760 --> 03:59:02,600 METRIC FOR EVERY SCREENING 5581 03:59:02,600 --> 03:59:08,640 ASSAY. IN THIS PIPELINE. SO ELY 5582 03:59:08,640 --> 03:59:10,440 LILLY MERCK AS WELL WHERE I USED 5583 03:59:10,440 --> 03:59:12,920 TO WORK IS REQUIRED METRIC 5584 03:59:12,920 --> 03:59:14,760 CALCULATED LIKE Z PRIME IS 5585 03:59:14,760 --> 03:59:19,200 CALCULATED MSR IS CALCULATED 5586 03:59:19,200 --> 03:59:20,160 BEFORE POSITION MADE SCREEN 5587 03:59:20,160 --> 03:59:24,880 COMPOUNDS WITH ASSAY. SO YOU 5588 03:59:24,880 --> 03:59:27,440 REALIZE FROM IN THE NEXT HALF 5589 03:59:27,440 --> 03:59:28,640 HOUR IT GETS DIRECTLY TO HEART 5590 03:59:28,640 --> 03:59:31,480 OF THE PROBLEM HOW WELL YOU CAN 5591 03:59:31,480 --> 03:59:32,520 DIFFERENTIATE POTENCY RESULTS 5592 03:59:32,520 --> 03:59:34,680 BETWEEN COMPOUNDS. SO Z PRIME 5593 03:59:34,680 --> 03:59:39,520 QUICK REVIEW, IT IS DIFFERENCE 5594 03:59:39,520 --> 03:59:40,920 BETWEEN SIGNAL AT TOP EXTREME 5595 03:59:40,920 --> 03:59:43,280 AND BOTTOM EXTREME HIGH 5596 03:59:43,280 --> 03:59:44,280 CONCENTRATION OF POSITIVE 5597 03:59:44,280 --> 03:59:48,360 CONTROL IN THE BACKGROUND. WE 5598 03:59:48,360 --> 03:59:50,440 CALCULATE VARIABILITY AROUND 5599 03:59:50,440 --> 03:59:53,000 THAT. THREE SDs CALCULATED AT 5600 03:59:53,000 --> 03:59:58,680 TOP AND BOTTOM. THIS GAP IS NUMB 5601 03:59:58,680 --> 04:00:00,840 RAY FOR OF Z PRIME FACTOR, 5602 04:00:00,840 --> 04:00:02,040 DENOMINATOR IS FULL WINDOW 5603 04:00:02,040 --> 04:00:04,080 SIGNALS TOP AND BOTTOM. SO THAT 5604 04:00:04,080 --> 04:00:06,000 IS YOUR Z PRIME. WHAT IS REALLY 5605 04:00:06,000 --> 04:00:07,680 IMPORTANT AND WHAT TC MIGHT NOT 5606 04:00:07,680 --> 04:00:10,240 HAVE TOUCHED UPON IS THAT THIS 5607 04:00:10,240 --> 04:00:12,480 VARIABILITY CALCULATE, STANDARD 5608 04:00:12,480 --> 04:00:13,840 DEVIATION CALCULATE IS HAS TO 5609 04:00:13,840 --> 04:00:14,920 TAKE INTO ACCOUNT ALL THE 5610 04:00:14,920 --> 04:00:18,720 SOURCES OF VARIABILITY IN YOUR 5611 04:00:18,720 --> 04:00:21,000 HDS. SO TYPICALLY WHEN Z PRIME 5612 04:00:21,000 --> 04:00:23,880 IS CALCULATED MOST PEOPLE 5613 04:00:23,880 --> 04:00:28,240 >>GENERATE DATA TOP AND BOTTOM 5614 04:00:28,240 --> 04:00:30,040 FROM SINGLE PLATE AND CALCULATE 5615 04:00:30,040 --> 04:00:31,080 THE MEAN AN STANDARD DEVIATION 5616 04:00:31,080 --> 04:00:35,640 FROM THE SINGLE PLATE. 5617 04:00:35,640 --> 04:00:36,960 CALCULATE Z PRIME FOR SINGLE 5618 04:00:36,960 --> 04:00:38,640 RATE. BUT THAT ONLY REFLECTS 5619 04:00:38,640 --> 04:00:41,800 VARIABILITY WITHIN THAT PLATE. 5620 04:00:41,800 --> 04:00:45,200 FOR HDS SCREENS YOU MAY NEED TO 5621 04:00:45,200 --> 04:00:46,400 WORRY ABOUT BETWEEN PLATE 5622 04:00:46,400 --> 04:00:46,960 VARIABILITY IF YOU ARE 5623 04:00:46,960 --> 04:00:51,600 UNDERSTANDING OVER TIME, IF SOME 5624 04:00:51,600 --> 04:00:56,480 PART OF THE HDS IS ONE LAB, WE 5625 04:00:56,480 --> 04:00:58,720 -- IF ROBOTICS ARE NOT INVOLVED 5626 04:00:58,720 --> 04:01:01,040 AS MUCH BUT MORE -- DEPENDING 5627 04:01:01,040 --> 04:01:03,000 UPON THE PARTICULAR TYPE OF HDS 5628 04:01:03,000 --> 04:01:04,440 AND DIFFERENT COMPONENTS OF 5629 04:01:04,440 --> 04:01:06,400 VARIABILITY INVOLVED IN THE HDS. 5630 04:01:06,400 --> 04:01:08,240 ALL THOSE VARIABILITY COMPONENTS 5631 04:01:08,240 --> 04:01:09,920 HAS TO BE INCORPORATED IN THE 5632 04:01:09,920 --> 04:01:14,680 CALCULATION OF THE VARIABILITY. 5633 04:01:14,680 --> 04:01:17,920 OTHERWISE WE OVERSTATE QUALITY 5634 04:01:17,920 --> 04:01:21,120 OF ASSAY, OVERESTIMATING Z PRIME 5635 04:01:21,120 --> 04:01:22,880 AND WE HAVE FALSE CONFIDENCE ON 5636 04:01:22,880 --> 04:01:25,800 THE ASSAY. T THAT Z PRIME VARIES 5637 04:01:25,800 --> 04:01:28,520 FROM .7 TO .4 DEPENDING HOW YOU 5638 04:01:28,520 --> 04:01:31,120 CALCULATE IT. IT IS IMPORTANT TO 5639 04:01:31,120 --> 04:01:32,360 MAKE SURE COMPONENTS OF 5640 04:01:32,360 --> 04:01:34,240 VARIABILITY IS RELEVANT FOR THAT 5641 04:01:34,240 --> 04:01:39,440 HDS INCO INCORPORATED IN Z PRIMO 5642 04:01:39,440 --> 04:01:41,880 THAT IS THE MESSAGE TO CONVEY 5643 04:01:41,880 --> 04:01:43,240 ABOUT Z PRIME BEFORE MOVING TO 5644 04:01:43,240 --> 04:01:45,840 THE OTHER TOPICS. ONE MORE 5645 04:01:45,840 --> 04:01:50,080 THING THAT ALSO SOMETHING TC 5646 04:01:50,080 --> 04:01:52,720 REVIEW, WE HAVE THREE ASSAYS AND 5647 04:01:52,720 --> 04:01:54,920 I MAKE THIS POINT THOUGH HE SAID 5648 04:01:54,920 --> 04:01:57,040 I WANT TO SAY AS WELL BECAUSE 5649 04:01:57,040 --> 04:01:58,560 THIS IS AN IMPORTANT THING AND I 5650 04:01:58,560 --> 04:02:02,720 SEE THE SAME MISTAKE BEING MADE 5651 04:02:02,720 --> 04:02:04,440 AFTER MANY YEARS NO MATTER HOW 5652 04:02:04,440 --> 04:02:06,400 MANY TIMES SAY IT PEOPLE USE 5653 04:02:06,400 --> 04:02:08,800 SIGNAL TO BACKGROUND ALSO SIGNAL 5654 04:02:08,800 --> 04:02:12,640 TO NOISE. AS WAY TO JUDGE 5655 04:02:12,640 --> 04:02:16,440 DEVELOP ASSAY OPTIMIZE ASSAY 5656 04:02:16,440 --> 04:02:17,880 CONDITIONS YOU SAW THIS POINT 5657 04:02:17,880 --> 04:02:20,760 MADE BY TC EARLIER. ASSAY ONE 5658 04:02:20,760 --> 04:02:22,880 EXCELLENT. SIGNAL TO BACKGROUND 5659 04:02:22,880 --> 04:02:24,600 OF TEN, SECOND IS EIGHT, THIRD 5660 04:02:24,600 --> 04:02:27,160 IS FIVE. SO GOING BY THAT PICK 5661 04:02:27,160 --> 04:02:29,080 THIS ASSAY CONDITION SAYING THIS 5662 04:02:29,080 --> 04:02:31,200 IS FANTASTIC, NICE WINDOW RATIO 5663 04:02:31,200 --> 04:02:33,000 OF TEN TO ONE BUT IF YOU TAKE 5664 04:02:33,000 --> 04:02:37,400 THE VARIABILITY INTO ACCOUNT, IT 5665 04:02:37,400 --> 04:02:41,200 IS ONE .4 FOR FIRST, .6 FOR 5666 04:02:41,200 --> 04:02:42,800 SECOND AND .75 FOR THE THIRD 5667 04:02:42,800 --> 04:02:50,440 ASSAY. IMPORTANT TO USE 5668 04:02:50,440 --> 04:02:53,080 APPROPRIATE METRIC WHEN WHAT 5669 04:02:53,080 --> 04:02:54,480 ASSAY CONDITIONS TO WORK WITH, 5670 04:02:54,480 --> 04:02:56,280 FORMAT TO WORK WITH, AND WHAT WE 5671 04:02:56,280 --> 04:02:58,520 ARE DEALING WITH, IN MAKING 5672 04:02:58,520 --> 04:03:00,080 POSITIONS ON IF WE START WITH 5673 04:03:00,080 --> 04:03:01,520 THE WRONG METRIC LIKE THIS 5674 04:03:01,520 --> 04:03:03,000 SIGNAL TO BACKGROUND WE END UP 5675 04:03:03,000 --> 04:03:04,120 WITH THE TOTALLY OPPOSITE 5676 04:03:04,120 --> 04:03:13,160 CONCLUSION. THIS PARTICULAR 5677 04:03:13,160 --> 04:03:14,200 CONTEXT YOU WANT TO TAKE 5678 04:03:14,200 --> 04:03:16,400 VARIABILITY INTO ACCOUNT, TO 5679 04:03:16,400 --> 04:03:17,840 CALCULATE Z PRIME USE IT TO 5680 04:03:17,840 --> 04:03:26,440 COMPARE THE DATA. ASP SINGLE 5681 04:03:26,440 --> 04:03:28,880 POINT HDS YOU ARE READY TO START 5682 04:03:28,880 --> 04:03:30,880 THE NEXT PACE OF SCREENING AND 5683 04:03:30,880 --> 04:03:32,480 CALCULATING WHAT THE POTENCY 5684 04:03:32,480 --> 04:03:37,640 METRIC S IC 50 KI SO FORTH, Z 5685 04:03:37,640 --> 04:03:39,400 PRIME IS NOT GOOD ENOUGH YOU CAN 5686 04:03:39,400 --> 04:03:41,200 HAVE Z PRIME .7 BUT WHAT DOES IT 5687 04:03:41,200 --> 04:03:43,560 MEAN IN TERMS OF HOW WELL WE CAN 5688 04:03:43,560 --> 04:03:45,400 DIFFERENTIATE THE POTENCY 5689 04:03:45,400 --> 04:03:46,680 VALUES? FOR THAT WE HAVE 5690 04:03:46,680 --> 04:03:50,440 SOMETHING CALLED THE MSR MINIMUM 5691 04:03:50,440 --> 04:03:54,280 SIGNIFICANT RATIO. SO BASICALLY 5692 04:03:54,280 --> 04:03:57,160 AS IT STATES BASICALLY SMALLEST 5693 04:03:57,160 --> 04:03:59,720 RATIO OR SMALLEST CHANGE ANY 5694 04:03:59,720 --> 04:04:01,440 POTENCY IN CELLS THAT YOU CAN 5695 04:04:01,440 --> 04:04:02,640 SEE IS STATISTICALLY 5696 04:04:02,640 --> 04:04:06,560 SIGNIFICANT. IF YOU HAVE TWO 5697 04:04:06,560 --> 04:04:10,480 COMPOUNDS WITH IC 5010 AND 20, 5698 04:04:10,480 --> 04:04:11,800 TWOFOLD DIFFERENT IS THAT 5699 04:04:11,800 --> 04:04:13,840 DIFFERENCE REAL? CAN YOU SAY 5700 04:04:13,840 --> 04:04:19,760 THAT THE COMPOUND TEN NANOMOLAR 5701 04:04:19,760 --> 04:04:22,280 IS SIGNIFICANTLY MORE POTENT 5702 04:04:22,280 --> 04:04:23,520 THAN 20 NANOMOLAR. THAT CEASE 5703 04:04:23,520 --> 04:04:25,800 WHAT MSL GETS AT. SO THE 5704 04:04:25,800 --> 04:04:28,040 FORMULA IS AS SHOWN HERE, 5705 04:04:28,040 --> 04:04:29,040 STRAIGHT FORWARD BUT CALCULATION 5706 04:04:29,040 --> 04:04:31,280 IS NOT SO STRAIGHT FORWARD SO WE 5707 04:04:31,280 --> 04:04:33,400 REVIEW THAT IN SOME DETAIL OVER 5708 04:04:33,400 --> 04:04:35,880 THE NEXT SLIDES. KEY PART OF 5709 04:04:35,880 --> 04:04:36,880 THE FORMULA IS STANDARD 5710 04:04:36,880 --> 04:04:39,240 DEVIATION YOU SEE THERE, SD. 5711 04:04:39,240 --> 04:04:41,920 THAT STANDARD DEVIATION IS 5712 04:04:41,920 --> 04:04:44,360 REPLICATE LOG IC 50 OR KI 5713 04:04:44,360 --> 04:04:48,760 WHATEVER THE METRIC IS. FROM 5714 04:04:48,760 --> 04:04:50,480 DIFFERENT SOURCES OF VARIABILITY 5715 04:04:50,480 --> 04:04:51,960 RELEVANT FOR SCREENING ASSAY. AT 5716 04:04:51,960 --> 04:04:53,680 THE COMMENT MADE ABOUT Z PRIME 5717 04:04:53,680 --> 04:04:55,240 HERE ALSO WE HAVE TAKE INTO 5718 04:04:55,240 --> 04:04:58,360 ACCOUNT INTRADAY INTERDAY 5719 04:04:58,360 --> 04:05:00,960 INTRALAB ANALYST ALL DIFFERENT 5720 04:05:00,960 --> 04:05:01,520 COMPONENTS RELEVANT FOR 5721 04:05:01,520 --> 04:05:02,960 PARTICULAR SCREENING ASSAY. ALL 5722 04:05:02,960 --> 04:05:05,520 THAT GOES INTO THIS CALCULATION 5723 04:05:05,520 --> 04:05:06,960 AND THERE ARE DIFFERENT WAYS TO 5724 04:05:06,960 --> 04:05:10,240 GO ABOUT IT SO WHAT DOES THAT 5725 04:05:10,240 --> 04:05:14,520 REALLY MEAN? MSR FOR ASSAY IS 5726 04:05:14,520 --> 04:05:17,080 FIVE. FOR ANY GIVEN COMPOUNDS 5727 04:05:17,080 --> 04:05:20,800 THAT MEANS THIS COMPOUND YOU 5728 04:05:20,800 --> 04:05:22,000 CANNOT BE SURE IT IS DIFFERENCE 5729 04:05:22,000 --> 04:05:23,920 FROM ALL THE OTHER COMPOUNDS 5730 04:05:23,920 --> 04:05:25,200 THAT HAVE ACTIVITY WITHIN FIVE 5731 04:05:25,200 --> 04:05:26,880 FOLD. OF THIS PARTICULAR 5732 04:05:26,880 --> 04:05:30,080 COMPOUND. FOR EXAMPLE, IC 50 5733 04:05:30,080 --> 04:05:33,320 PARTICULAR COMPOUND IS 100, THEN 5734 04:05:33,320 --> 04:05:36,280 WE DON'T HAVE 95% CONFIDENCE TO 5735 04:05:36,280 --> 04:05:37,680 SAY THIS COMPOUND IS DIFFERENCE 5736 04:05:37,680 --> 04:05:39,400 FROM ANY OTHER COMPOUND THAT HAS 5737 04:05:39,400 --> 04:05:43,360 IC 50 BETWEEN 20 AND 500. 100 5738 04:05:43,360 --> 04:05:45,280 DIVIDED BY FIVE, 100 TIMES FIVE 5739 04:05:45,280 --> 04:05:47,880 SO WILL IS AFIELD FOLD GRAY ZONE 5740 04:05:47,880 --> 04:05:51,440 FOR ANY GIVEN COMPOUNDS. THAT 5741 04:05:51,440 --> 04:05:54,560 IS HUGE, THREEFOLD CANNING BIG 5742 04:05:54,560 --> 04:05:56,320 DEPENDING ON OBJECTIVE OF THE 5743 04:05:56,320 --> 04:05:58,880 SCREENING ASSAY. THIS IS WHY I 5744 04:05:58,880 --> 04:06:00,040 SAID GETS DIRECTLY TO HEART OF 5745 04:06:00,040 --> 04:06:02,760 THE PROBLEM WHICH IS HOW WELL 5746 04:06:02,760 --> 04:06:05,320 HOW CONFIDENT AM ID IN SAYING 5747 04:06:05,320 --> 04:06:06,280 ONE IS BETTER THAN THE OTHER AND 5748 04:06:06,280 --> 04:06:08,160 HOW CONFIDENT AM I IN RANK 5749 04:06:08,160 --> 04:06:09,640 ORDERING THESE COMPOUNDS BASED 5750 04:06:09,640 --> 04:06:12,640 ON THIS DATA. SO THAT IS WHY MSR 5751 04:06:12,640 --> 04:06:13,520 IS AT THAT PARTICULAR CORE 5752 04:06:13,520 --> 04:06:16,720 QUESTION. OBJECTIVE OF SCREENING 5753 04:06:16,720 --> 04:06:19,320 DATA. Z PRIME DOESN'T. SO Z 5754 04:06:19,320 --> 04:06:20,600 PRIME TELLS YOU DIFFERENTIATE 5755 04:06:20,600 --> 04:06:22,400 ACTIVE FROM INACTIVE, WE HAVE 5756 04:06:22,400 --> 04:06:23,480 GONE PAST THAT WHEN YOU ARE 5757 04:06:23,480 --> 04:06:25,160 LOOKING AT POTENCY DATA AND 5758 04:06:25,160 --> 04:06:28,760 MAKING COMPARISONS IN TERMS OF 5759 04:06:28,760 --> 04:06:33,040 POTENCY VALUES. THAT IS WHY THIS 5760 04:06:33,040 --> 04:06:34,560 MSR READS SO THE QUESTION WE GET 5761 04:06:34,560 --> 04:06:37,120 IS CALCULATE MSR, WHAT ARE WE, 5762 04:06:37,120 --> 04:06:43,080 TWO, THREE, FIVE, WHATEVER? LIKE 5763 04:06:43,080 --> 04:06:45,680 Z PRIME OF .7 OR 1 POINT 3, YOU 5764 04:06:45,680 --> 04:06:47,040 SAY METRIC THAT REFLECT IT IS 5765 04:06:47,040 --> 04:06:48,280 QUALITY OF HDS FROM ALL THE 5766 04:06:48,280 --> 04:06:49,840 DIFFERENT COMPOUNDS THAT HAVE 5767 04:06:49,840 --> 04:06:52,520 TESTED IN HDS. SIMILARLY, MSR WE 5768 04:06:52,520 --> 04:06:54,520 ARE SAYING WHAT ARE WE 5769 04:06:54,520 --> 04:06:56,600 CALCULATING IT OUT TO BE, 2, 3, 5770 04:06:56,600 --> 04:06:59,600 5, THAT VALUE REPRESENTS 5771 04:06:59,600 --> 04:07:03,160 ROBUSTNESS OF DIFFERENTIATING 5772 04:07:03,160 --> 04:07:05,840 ANY TWO POTENCY RISK SALES FOR 5773 04:07:05,840 --> 04:07:07,560 ANY PAIR OF COMPOUNDS, HIGH LIPO 5774 04:07:07,560 --> 04:07:10,440 TENT COMPOUND OR EXTREME LIPO 5775 04:07:10,440 --> 04:07:13,200 TEN OR LESS POE TEN COMPOUNDS. 5776 04:07:13,200 --> 04:07:16,120 HOW CAN YOU SAY THAT WHEN 5777 04:07:16,120 --> 04:07:17,720 VARIABILITY OF IC 50 DEPENDS ON 5778 04:07:17,720 --> 04:07:22,880 IC 50. THIS IS A GRAPH HERE IS 5779 04:07:22,880 --> 04:07:24,920 IC 50 OF BATCH OF COMPOUNDS 5780 04:07:24,920 --> 04:07:28,560 AGAINST VARIABILITY OF IC 50. 5781 04:07:28,560 --> 04:07:30,840 YOU CAN SEE HOW VARIABILITY IS 5782 04:07:30,840 --> 04:07:32,320 NOT CONSISTENT ACROSS THE RANGE 5783 04:07:32,320 --> 04:07:35,080 OF COMPOUNDS. THIS IS TYPICAL, 5784 04:07:35,080 --> 04:07:37,800 THIS IS NOT THIS PARTICULAR 5785 04:07:37,800 --> 04:07:39,600 ASSAY ALONE. THIS IS NOT ANY 5786 04:07:39,600 --> 04:07:44,080 ASSAY WHEN YOU CORRELATE 5787 04:07:44,080 --> 04:07:46,240 COMPOUND VARIABILITY OF IC 50 5788 04:07:46,240 --> 04:07:47,920 THIS IS TREND YOU SEE, 5789 04:07:47,920 --> 04:07:49,440 VARIABILITY GOES UP AS IC 50 5790 04:07:49,440 --> 04:07:51,600 GOES UP SO CALCULATE A SINGLE 5791 04:07:51,600 --> 04:07:52,880 METRIC LIKE MV THAT REPRESENTS 5792 04:07:52,880 --> 04:07:56,880 ALL THESE COMPOUNDS. THE ANSWER 5793 04:07:56,880 --> 04:08:01,200 LIES HERE. IN THE LOG SCALE, 5794 04:08:01,200 --> 04:08:03,400 VARIABILITY IS UNIFORM. THIS IS 5795 04:08:03,400 --> 04:08:06,600 THE SAME DATA BUT IT IS MEAN LOG 5796 04:08:06,600 --> 04:08:08,120 IC 50 AGAINST VARIABILITY 5797 04:08:08,120 --> 04:08:11,160 STANDARD DEVIATION OF LOG IC 50. 5798 04:08:11,160 --> 04:08:13,160 YOU CAN SEE CONSISTENT ACROSS 5799 04:08:13,160 --> 04:08:15,800 ENTIRE RANGE. THIS IS TRUE FOR 5800 04:08:15,800 --> 04:08:18,400 ANY POTENCY DATA, ANY MEASURE, 5801 04:08:18,400 --> 04:08:22,120 CANI IC 50 EC 50. SO THAT IS WHY 5802 04:08:22,120 --> 04:08:24,800 ANALYSIS IS DONE IN TERMS OF THE 5803 04:08:24,800 --> 04:08:27,600 LOG POTENCY VALUES. THAT IS 5804 04:08:27,600 --> 04:08:29,680 WHAT MAKES THIS METRIC 5805 04:08:29,680 --> 04:08:31,800 REPRESENTATIVE OF ALL THE 5806 04:08:31,800 --> 04:08:32,520 COMPOUNDS FROM THAT PARTICULAR 5807 04:08:32,520 --> 04:08:38,360 ASSAY. AND WE HAVE Z PRIME 5808 04:08:38,360 --> 04:08:39,600 REPRESENTS THE BILLIONS OF 5809 04:08:39,600 --> 04:08:41,400 COMPOUNDS TESTED IN HDS, WHEN 5810 04:08:41,400 --> 04:08:43,720 YOU CALCULATE REPRESENTS ALL THE 5811 04:08:43,720 --> 04:08:44,720 COMPOUNDS, HUNDREDS OF THOUSANDS 5812 04:08:44,720 --> 04:08:45,800 OF COMPOUNDS TESTED IN THAT 5813 04:08:45,800 --> 04:08:49,000 SCREENING ASSAY. SO THEREFORE 5814 04:08:49,000 --> 04:08:53,840 MSR CALCULATIONS IS BASED ON LOG 5815 04:08:53,840 --> 04:08:56,520 POTENCY INSTEAD OF ORIGINAL TO 5816 04:08:56,520 --> 04:08:57,880 ENSURE VARIABILITY ACROSS ALL 5817 04:08:57,880 --> 04:08:59,440 COMPOUNDS, THE FORMULA IS 5818 04:08:59,440 --> 04:09:01,480 DERIVED FROM THE 95 PURCHASE 5819 04:09:01,480 --> 04:09:04,360 CONFIDENCE INTERVAL CONCEPT. SO 5820 04:09:04,360 --> 04:09:05,280 95% CONFIDENCE INTERVAL FOR 5821 04:09:05,280 --> 04:09:07,280 DIFFERENCE OF ANY TWO LOG 5822 04:09:07,280 --> 04:09:08,960 POTENCIES IS BASICALLY LOOKS 5823 04:09:08,960 --> 04:09:13,560 LIKE THIS. THIS IS Y 1 MINUS Y 2 5824 04:09:13,560 --> 04:09:16,160 DIFFERENCE OF TWO LOG IC 50s, 5825 04:09:16,160 --> 04:09:18,120 TO DEFINE 95% CONFIDENCE 5826 04:09:18,120 --> 04:09:19,560 INTERVAL FOR THE DIFFERENCE OF 5827 04:09:19,560 --> 04:09:22,000 LOG POTENCIES. THIS FACTOR IS 5828 04:09:22,000 --> 04:09:24,800 WHAT WE CALL MINIMUM SIGNIFICANT 5829 04:09:24,800 --> 04:09:26,440 DIFFERENCE OF THE LOG POTENCY OF 5830 04:09:26,440 --> 04:09:30,440 THE TWO COMPOUNDS. IF YOU TAKE 5831 04:09:30,440 --> 04:09:34,760 THE ANTI-OF THAT, THAT GIVES YOU 5832 04:09:34,760 --> 04:09:36,720 MSR. SO DIFFERENCE OF LOG IS LOG 5833 04:09:36,720 --> 04:09:39,200 RATIO. YOU TAKE ANTI-LOG OF 5834 04:09:39,200 --> 04:09:40,520 THAT, THAT'S THE RATIO. THAT IS 5835 04:09:40,520 --> 04:09:46,360 WHY YOU HAVE THE RATIO. WHY MRS 5836 04:09:46,360 --> 04:09:48,920 INSTEAD OF MSD. IS THAT CLEAR? 5837 04:09:48,920 --> 04:09:51,480 OKAY. SO WHAT CRITERIA DO WE 5838 04:09:51,480 --> 04:09:53,240 HAVE TO APPLY HERE? BASED ON MY 5839 04:09:53,240 --> 04:09:55,480 EXPERIENCE, TALKING WITH VARIOUS 5840 04:09:55,480 --> 04:09:58,040 CHEMISTS AND BIOLOGISTS OVER 5841 04:09:58,040 --> 04:09:59,160 YEARS AND DIFFERENCE DISCOVERY 5842 04:09:59,160 --> 04:10:00,040 PROGRAMS, I FOUND GENERALLY 5843 04:10:00,040 --> 04:10:03,280 PEOPLE ARE HAPPY, IF LESS THAN 5844 04:10:03,280 --> 04:10:06,480 THREE. NO QUESTIONS ASKED 5845 04:10:06,480 --> 04:10:07,920 USUALLY. WHAT EVEN IS GREATER 5846 04:10:07,920 --> 04:10:09,720 THAN THREE THEY START GETTING 5847 04:10:09,720 --> 04:10:10,880 CONCERNED AND THEN DEPENDS ON 5848 04:10:10,880 --> 04:10:12,120 CONTEXT. IF YOU ARE DEALING WITH 5849 04:10:12,120 --> 04:10:13,920 A BROAD RANGE OF ACTORS OR THIS 5850 04:10:13,920 --> 04:10:15,120 PARTICULAR ASSAY IS PROFILING 5851 04:10:15,120 --> 04:10:18,880 ASSAY, NOT A PRIMARY ASSAY, 5852 04:10:18,880 --> 04:10:24,600 SLIGHTLY HIGHER VALUE OF MSR 5853 04:10:24,600 --> 04:10:25,800 3.54.5 IS ACCEPTABLE IN THOSE 5854 04:10:25,800 --> 04:10:27,440 CASES BUT IF THAT IS TOO HIGH, 5855 04:10:27,440 --> 04:10:29,160 IF YOU DON'T HAVE TIME TO 5856 04:10:29,160 --> 04:10:33,040 REOPTIMIZE THE ASSAY, ONE 5857 04:10:33,040 --> 04:10:34,480 SOLUTION IS SCREEN COMPOUNDS 5858 04:10:34,480 --> 04:10:36,720 MORE THAN ONCE. THAT 5859 04:10:36,720 --> 04:10:38,320 AUTOMATICALLY REDUCES 5860 04:10:38,320 --> 04:10:40,320 VARIABILITY AND THEREFORE MSR OF 5861 04:10:40,320 --> 04:10:43,120 SAY FOUR MAY COME DOWN TO 2.5 IF 5862 04:10:43,120 --> 04:10:47,120 SCREENING ALL COMPOUND TWICE. 5863 04:10:47,120 --> 04:10:48,360 IF LOW THROUGH PUT ASSAY, CELL 5864 04:10:48,360 --> 04:10:49,920 BASED ASSAY, TAKES A LONG TIME 5865 04:10:49,920 --> 04:10:54,920 TO GENERATE THE DATA, WE CANNOT 5866 04:10:54,920 --> 04:10:57,360 SCREEN COMPOUNDS TWICE OR TWO OR 5867 04:10:57,360 --> 04:10:59,480 THREE TIMES THROUGH PUT SO THAT 5868 04:10:59,480 --> 04:11:02,400 CASE BEST SOLUTION IS REOPTIMIZE 5869 04:11:02,400 --> 04:11:06,720 TO IMPROVE VARIABILITY SO BOTH 5870 04:11:06,720 --> 04:11:12,160 SCENARIOS IN SOME THEY SAY QUICK 5871 04:11:12,160 --> 04:11:13,080 SCREEN AT LEAST SUBSET OF 5872 04:11:13,080 --> 04:11:15,280 COMPOUNDS MORE THAN ONCE AND GET 5873 04:11:15,280 --> 04:11:16,680 MSR LOW AND SATISFACTORY AND 5874 04:11:16,680 --> 04:11:19,240 MOVE ON. I HAVE ALSO SEEN THE 5875 04:11:19,240 --> 04:11:20,880 OTHER EXTREME, THE CELL BASED 5876 04:11:20,880 --> 04:11:23,680 ASSAY TAKES A LONG TIME GENERATE 5877 04:11:23,680 --> 04:11:25,680 SO BACK TO DRAWING BOARD SPEND 5878 04:11:25,680 --> 04:11:27,920 WEEKS TO REOPTIMIZE, BROUGHT 5879 04:11:27,920 --> 04:11:31,560 VARIABILITY DOWN AND MSR CAME 5880 04:11:31,560 --> 04:11:34,800 FROM POSITIVE 6 WHATEVER TO 2.5, 5881 04:11:34,800 --> 04:11:36,800 2.7 AFTER OPTIMIZE ASSAY OR 5882 04:11:36,800 --> 04:11:38,640 CHANGE FORMAT AND CELL LINES AND 5883 04:11:38,640 --> 04:11:39,760 DEPENDING ON THE CONTEXT YOU 5884 04:11:39,760 --> 04:11:41,280 HAVE TO WORK AROUND WITH IT TO 5885 04:11:41,280 --> 04:11:46,600 GET THE DATA YOU WANT. THIS IS 5886 04:11:46,600 --> 04:11:48,480 WHAT HAPPENS IF YOU SCREEN MORE 5887 04:11:48,480 --> 04:11:51,520 THAN ONCE, IF YOU GET FROM A 5888 04:11:51,520 --> 04:11:53,480 SINGLE SCREEN, IF YOU SCREEN 5889 04:11:53,480 --> 04:11:55,040 COMPOUNDS MORE THAN ONCE SAY 5890 04:11:55,040 --> 04:11:57,560 TWICE, AUTOMATICALLY COMES DOWN 5891 04:11:57,560 --> 04:12:02,600 TO 3.96 FROM 7 TO 4. SCREEN 5892 04:12:02,600 --> 04:12:04,080 THREE TIMES COMES DOWN TO THREE 5893 04:12:04,080 --> 04:12:06,240 AND PLATEAUS AFTER A WHILE. SO 5894 04:12:06,240 --> 04:12:08,480 NOT MUCH GAIN AFTER CERTAIN 5895 04:12:08,480 --> 04:12:19,000 WHILE. COMES DOWN TO 2.2, THEN 2 5896 04:12:19,000 --> 04:12:21,160 THEN PLATEAUS. YOU CAN HAVE MSR 5897 04:12:21,160 --> 04:12:23,080 OF TEN BUT COMPOUNDS FOUR TIMES 5898 04:12:23,080 --> 04:12:27,720 BRING IT DOWN TO ALMOST THREE. 5899 04:12:27,720 --> 04:12:29,040 DOESN'T MEAN SCREEN ALL 5900 04:12:29,040 --> 04:12:30,400 COMPOUNDS THAT MANY TIMES ONLY 5901 04:12:30,400 --> 04:12:32,240 ONES THAT ARE INTEREST TO YOU 5902 04:12:32,240 --> 04:12:33,920 THAT MEET ACTIVITY CRITERIA, IF 5903 04:12:33,920 --> 04:12:38,160 IT IS ONE MICROMOLAR MSR IS 5904 04:12:38,160 --> 04:12:40,840 FOUR, YOU CAN SCREEN ALL THOSE 5905 04:12:40,840 --> 04:12:43,320 COMPOUNDS THAT HAVE/LESS THAN 5906 04:12:43,320 --> 04:12:45,640 FOUR MICROMOLAR MORE THAN ONCE. 5907 04:12:45,640 --> 04:12:48,240 SO FOR THOSE COMPOUNDS, ACTIVITY 5908 04:12:48,240 --> 04:12:50,480 LESS THAN 4 MICROMOLAR YOU CAN 5909 04:12:50,480 --> 04:12:57,040 -- IF YOU SCREEN MORE THAN ONCE, 5910 04:12:57,040 --> 04:12:59,240 MSR 2.7 TO INTERPRET THE DATA 5911 04:12:59,240 --> 04:13:04,320 FROM THOSE COMPOUNDS. THE KEY 5912 04:13:04,320 --> 04:13:06,360 PART OF THE FORMULA HERE IS 5913 04:13:06,360 --> 04:13:08,360 DIVIDING BY SQUARE ROOT OF N. 5914 04:13:08,360 --> 04:13:11,800 THIS N HERE IS NUMBER OF TIMES 5915 04:13:11,800 --> 04:13:13,720 SCREEN COMPOUNDS. THAT IS WHY 5916 04:13:13,720 --> 04:13:17,160 ABLE CALCULATE ALL THE SCENAR 5917 04:13:17,160 --> 04:13:19,160 SCENARIOS. SO PLUCK THE VALUE N 5918 04:13:19,160 --> 04:13:21,200 OF TWO, THREE, FOUR, WHATEVER N 5919 04:13:21,200 --> 04:13:22,680 YOU LIKE AND THAT IS HOW YOU 5920 04:13:22,680 --> 04:13:27,280 GENERATE MSR VALUES. SO YOU CAN 5921 04:13:27,280 --> 04:13:28,280 SIGH BEFORE YOU SCREEN COMPOUNDS 5922 04:13:28,280 --> 04:13:30,280 YOU CAN GET IDEA WHAT MSR AND 5923 04:13:30,280 --> 04:13:32,920 ALSO MAKE A DECISION HOW MANY 5924 04:13:32,920 --> 04:13:33,480 TIMES YOU SHOULD SCREEN 5925 04:13:33,480 --> 04:13:38,680 COMPOUNDS TO GET ACCEPTABLE MSR 5926 04:13:38,680 --> 04:13:40,400 SO GREAT WAY TO FIGURE OUT HOW 5927 04:13:40,400 --> 04:13:41,520 MANY TIMES SHOULD I DO IT. I 5928 04:13:41,520 --> 04:13:44,400 HAVE SEEN THE SCENARIO ONCE BY 5929 04:13:44,400 --> 04:13:46,080 DEFAULT THE PROGRAM DISCOVERY 5930 04:13:46,080 --> 04:13:48,640 PROGRAM WAS ASKED TO SCREEN ALL 5931 04:13:48,640 --> 04:13:51,400 THE COMPOUNDS THREE TIME, 5932 04:13:51,400 --> 04:13:54,400 WITHOUT ANY RATIONALE, OBJECTIVE 5933 04:13:54,400 --> 04:13:56,640 RATIONALE, JUST EXPERIENCE, 5934 04:13:56,640 --> 04:14:01,160 BIOLOGIES DO IT THREE TIMES. 5935 04:14:01,160 --> 04:14:02,360 BIOLOGISTS SCREEN COMPOUNDS 5936 04:14:02,360 --> 04:14:05,160 DIDN'T FEEL GOOD, CAME TO ME AND 5937 04:14:05,160 --> 04:14:07,960 SAID THIS DATA IS ROBUST, BUT I 5938 04:14:07,960 --> 04:14:08,920 SUGGEST ALL COMPOUNDS THREE 5939 04:14:08,920 --> 04:14:10,280 TIMES DOES IT MAKE SENSE, COME 5940 04:14:10,280 --> 04:14:14,040 UP WITH RATIONALE FOR THAT? SO 5941 04:14:14,040 --> 04:14:17,640 WE CALCULATED THE MSR, TO THE N 5942 04:14:17,640 --> 04:14:20,200 OF ONE, SCREEN COMPOUNDS ONLY 5943 04:14:20,200 --> 04:14:26,280 ONE, MSR IS 2.1. IF YOU SCREEN 5944 04:14:26,280 --> 04:14:30,240 TWICE IT COMES DOWN TO 1.8, 1 5945 04:14:30,240 --> 04:14:33,640 POINT THREE TIMES, AROUND 1.6, 5946 04:14:33,640 --> 04:14:37,960 SO NOT MUCH TO BE GAINED FINAL 5947 04:14:37,960 --> 04:14:38,880 SCREENING ONCE VERSUS WISE 5948 04:14:38,880 --> 04:14:41,320 TWICE, CERTAINLY NOT MUCH GAINED 5949 04:14:41,320 --> 04:14:43,160 FROM THREE TIME TIMES. SO HE USS 5950 04:14:43,160 --> 04:14:47,920 DATA TO CONVINCE COLLEAGUES TO 5951 04:14:47,920 --> 04:14:50,760 SAY LOOK GUYS REDUCING THROUGH 5952 04:14:50,760 --> 04:14:52,840 PUT THREEFOLD GENERATING THIS 5953 04:14:52,840 --> 04:14:54,680 DATA OVER THREE REPLICATES. 5954 04:14:54,680 --> 04:14:58,680 LET'S DO IT ONCE, AND LIMIT 5955 04:14:58,680 --> 04:15:02,200 SECOND TESTING TO ONLY THOSE 5956 04:15:02,200 --> 04:15:04,080 COMPOUNDS INTERESTING SAY LESS 5957 04:15:04,080 --> 04:15:08,560 TAN 100 NANOMOLAR ACTIVITY. REST 5958 04:15:08,560 --> 04:15:10,320 OF THE TEAM AGREED. THIS IS HOW 5959 04:15:10,320 --> 04:15:11,920 YOU GO ABOUT JUSTIFYING, ANOTHER 5960 04:15:11,920 --> 04:15:17,000 QUESTION THAT COMES UP IS DO TEN 5961 04:15:17,000 --> 04:15:20,720 POINT DOSE RESPONSE CURVE, 8 5962 04:15:20,720 --> 04:15:21,520 POINT, 12 POINT DOSE RESPONSE 5963 04:15:21,520 --> 04:15:23,080 CURVE? ? YOU CAN GENERATE DATA 5964 04:15:23,080 --> 04:15:24,080 BEFORE YOU SCREEN THE COMPOUND 5965 04:15:24,080 --> 04:15:26,480 YOU CAN GENERALOR GENERATE -- 5966 04:15:26,480 --> 04:15:28,800 RUN A BATCH OF COMPOUNDS USING 5967 04:15:28,800 --> 04:15:31,240 DIFFERENT CONCENTRATIONS, 8 5968 04:15:31,240 --> 04:15:33,480 POINT FORMAT, 15 POINT FORMAT 5969 04:15:33,480 --> 04:15:35,840 AND CALCULATE MSR FOR THE 5970 04:15:35,840 --> 04:15:37,880 DIFFERENT SCENARIOS. THE M SR 5971 04:15:37,880 --> 04:15:40,480 FOR THE 8 POINT FORMAT IS GOOD 5972 04:15:40,480 --> 04:15:43,480 ENOUGH YOU DON'T HAVE TO OVERDO 5973 04:15:43,480 --> 04:15:49,960 IT. THIS CONCEPT IS POWERFUL FOR 5974 04:15:49,960 --> 04:15:51,040 THESE POSITIONS EARLY ON AS 5975 04:15:51,040 --> 04:15:55,560 WELL. BUT OF COURSE LATER ON 5976 04:15:55,560 --> 04:16:01,040 THIS IS CONTROL CONTROL TRICK TO 5977 04:16:01,040 --> 04:16:02,400 KEEP CONTROL OF THE QUALITY OF 5978 04:16:02,400 --> 04:16:04,240 THE ASSAY. SO DIFFERENT WAYS OF 5979 04:16:04,240 --> 04:16:06,160 ESTIMATING, ONE IS TEST FREE 5980 04:16:06,160 --> 04:16:11,080 TEST MSR. BEFORE YOU SCREEN 5981 04:16:11,080 --> 04:16:13,680 COMPOUNDS YOU CAN RUN IT TWICE 5982 04:16:13,680 --> 04:16:15,040 AND THESE CAN BE SOME TEST 5983 04:16:15,040 --> 04:16:16,960 COMPOUNDS THAT YOU HAVE, YOU 5984 04:16:16,960 --> 04:16:20,240 HAVE FOUR OR FIVE COMPOUNDS YOU 5985 04:16:20,240 --> 04:16:23,120 CAN WORK WITH YOU CAN CREATE 5986 04:16:23,120 --> 04:16:25,800 REPLY CASE OF COMPOUNDS AND 5987 04:16:25,800 --> 04:16:26,600 PRE-SELECT DIFFERENT COMPOUNDS 5988 04:16:26,600 --> 04:16:29,440 SO YOU HAVE 20 OR 30 COMPOUNDS 5989 04:16:29,440 --> 04:16:31,040 FROM 405 COMPOUNDS AND TEST THEM 5990 04:16:31,040 --> 04:16:34,760 OVER TWO RUNS AND GENERATE -- 5991 04:16:34,760 --> 04:16:39,800 USE THOSE DATA TO CALCULATE TEST 5992 04:16:39,800 --> 04:16:42,840 FREE M,R AND THAT GIVES YOU A 5993 04:16:42,840 --> 04:16:44,320 GOOD FEEL HOW IT WILL BE DOWN 5994 04:16:44,320 --> 04:16:45,960 THE ROAD, AND IF GOOD MUST HAVE 5995 04:16:45,960 --> 04:16:49,920 YOU CAN SCREEN COMPOUNDS IN THE 5996 04:16:49,920 --> 04:16:53,560 FULL MODE. THEN YOU HAVE A 5997 04:16:53,560 --> 04:16:56,600 MOVING MSR BASED ON POSITIVE 5998 04:16:56,600 --> 04:16:58,560 CONTROL AND WE HAVE 5999 04:16:58,560 --> 04:17:00,520 RETROSPECTIVE MSR WHERE WE USE 6000 04:17:00,520 --> 04:17:02,400 ALL THE DATA TESTED AND 6001 04:17:02,400 --> 04:17:03,960 COMPOUNDS TESTED FOR PERIOD OF 6002 04:17:03,960 --> 04:17:05,800 TIME INCLUDING THE POSITIVE 6003 04:17:05,800 --> 04:17:08,280 CONTROL AS WELL AS ALL TEST 6004 04:17:08,280 --> 04:17:09,920 COMPOUNDS YOU RUN OVER PERIOD OF 6005 04:17:09,920 --> 04:17:11,960 TIME AND THEN CALCULATE WHAT MSR 6006 04:17:11,960 --> 04:17:18,080 IS FOR ALL THOSE DATA. SCENARIOS 6007 04:17:18,080 --> 04:17:19,560 DEPENDING ON THE APPLICATION. 6008 04:17:19,560 --> 04:17:22,000 TEST FREE TEST AS MENTIONED 6009 04:17:22,000 --> 04:17:23,720 EARLIER, SMALL BATCH OF 6010 04:17:23,720 --> 04:17:26,080 COMPOUNDS TESTED TWICE, CAPTURES 6011 04:17:26,080 --> 04:17:33,000 WITHIN RUN VARIABILITY BECAUSE 6012 04:17:33,000 --> 04:17:34,160 TWO RUNS BUT EARLY FILTER TO 6013 04:17:34,160 --> 04:17:36,280 MAKE A DECISION WHETHER TO START 6014 04:17:36,280 --> 04:17:38,800 FULL SCREENING OF COMPOUNDS. 6015 04:17:38,800 --> 04:17:40,880 BECAUSE IF THIS MSR IS BAD THERE 6016 04:17:40,880 --> 04:17:43,400 IS NO POINT STARTING THE SCREEN. 6017 04:17:43,400 --> 04:17:46,880 IF YOU START THE SCREEN YOU HAVE 6018 04:17:46,880 --> 04:17:48,320 INTERPLATE VARIABILITY AND IT 6019 04:17:48,320 --> 04:17:52,400 WILL GET WORS WORSE. O SO THIS A 6020 04:17:52,400 --> 04:17:54,560 QUICK FOR DECIDING WHETHER START 6021 04:17:54,560 --> 04:18:00,440 SCREENING OR NOT. THERE IS A 6022 04:18:00,440 --> 04:18:01,680 TOOL AVAILABLE FOR CALCULATING 6023 04:18:01,680 --> 04:18:04,000 THAT. HERE IS OUTPUT FROM TEST 6024 04:18:04,000 --> 04:18:05,160 FREE TEST DATA SO WHAT WE HAVE 6025 04:18:05,160 --> 04:18:07,920 IS RATIO OF POTENCY RESULTS 6026 04:18:07,920 --> 04:18:09,760 AGAINST THE AVERAGE POTENCY. AND 6027 04:18:09,760 --> 04:18:11,520 THIS IS A MEAN RATIO OF ALL 6028 04:18:11,520 --> 04:18:14,720 THOSE COMPOUNDS THAT WERE 6029 04:18:14,720 --> 04:18:15,800 TESTED, 95% CONFIDENCE INTERVAL 6030 04:18:15,800 --> 04:18:17,120 WITH LIMITS OF AGREEMENT 6031 04:18:17,120 --> 04:18:22,320 PROVIDED AND MSR VALUE 8.54. H 6032 04:18:22,320 --> 04:18:24,440 IS 3.54 HERE MOSTLY WITHIN 6033 04:18:24,440 --> 04:18:26,880 VARIABILITY. IF YOU -- IF YOU 6034 04:18:26,880 --> 04:18:28,160 WON'T FOLLOW-UP WITH SCREEN AN 6035 04:18:28,160 --> 04:18:29,400 SCREEN ALL YOUR COMPOUNDS WITH 6036 04:18:29,400 --> 04:18:30,680 THIS ASSAY, IT IS GOING TO GET 6037 04:18:30,680 --> 04:18:39,360 WORSE. SO YOU MAY GO BACK TO 6038 04:18:39,360 --> 04:18:41,840 IMPROVE ASSAY BEFORE YOU SCREEN 6039 04:18:41,840 --> 04:18:46,320 COMPOUNDS THIS IS GREAT FILTER 6040 04:18:46,320 --> 04:18:50,240 FOR DECIDING WHETHER TO MOVE 6041 04:18:50,240 --> 04:18:52,040 FORWARD OR NOT WITH THE SCREEN 6042 04:18:52,040 --> 04:18:57,040 BEFORE YOU REALIZE TOO LATE. YOU 6043 04:18:57,040 --> 04:18:59,240 CAN USE THE DATA TO DECIDE HOW 6044 04:18:59,240 --> 04:19:02,000 MANY TIMES TEST COMPOUNDS, SO 6045 04:19:02,000 --> 04:19:03,280 THIS 3.54 BUZZ BASED ON SINGLE 6046 04:19:03,280 --> 04:19:08,600 RUN OF COMPOUNDS, IF YOU ARE -- 6047 04:19:08,600 --> 04:19:09,720 DECIDE TO TEST COMPOUNDS TWICE 6048 04:19:09,720 --> 04:19:13,200 YOU CAN PLUG IN THE VALUE IN 6049 04:19:13,200 --> 04:19:16,480 THAT FORMULA I SHOWED YOU 6050 04:19:16,480 --> 04:19:18,600 EARLIER, PLUG IN THE VALUE N 6051 04:19:18,600 --> 04:19:22,120 EQUAL TO TWO AND 3.54 COME DOWN 6052 04:19:22,120 --> 04:19:23,960 TO 2.5. SO IF THAT IS WHAT YOU 6053 04:19:23,960 --> 04:19:25,840 DECIDE TO DO, THAT IS REASONABLE 6054 04:19:25,840 --> 04:19:28,240 AND YOU MIGHT SAY, AT LEAST FOR 6055 04:19:28,240 --> 04:19:29,680 THOSE COMPOUNDS THAT MEET 6056 04:19:29,680 --> 04:19:34,760 ACTIVITY CRITERIA, SCREEN THEM 6057 04:19:34,760 --> 04:19:37,320 TWICE, ACCEPTABLE MSR 2.5 AND 6058 04:19:37,320 --> 04:19:38,520 SEE HOW IT WORKS OUT DURING YOUR 6059 04:19:38,520 --> 04:19:43,440 SCREEN. QUICK ILLUSTRATION HOW 6060 04:19:43,440 --> 04:19:46,200 TO CALCULATE IT SO TEST FREE 6061 04:19:46,200 --> 04:19:48,360 TEST DATA BATCH OF COMPOUNDS 6062 04:19:48,360 --> 04:19:50,480 TEST TWICE, TAKE LOG VALUES OF 6063 04:19:50,480 --> 04:19:51,400 THOSE TWO. TAKE DIFFERENCE OF 6064 04:19:51,400 --> 04:19:55,680 THE LOGS. THEN TAKE STANDARD 6065 04:19:55,680 --> 04:19:57,200 DEVIATION OF DIFFERENCES OF THE 6066 04:19:57,200 --> 04:19:59,440 LOGS. AND PLUG INTO THE FORMULA 6067 04:19:59,440 --> 04:20:06,120 AND GET THE MSR. THIS CONCEPT 6068 04:20:06,120 --> 04:20:07,440 OF COMPARISON BATCH OF COMPOUNDS 6069 04:20:07,440 --> 04:20:09,760 IS USEFUL WHEN TRANSFERRING THE 6070 04:20:09,760 --> 04:20:15,320 ASSAY FROM ONE LAB TO ANOTHER, A 6071 04:20:15,320 --> 04:20:16,800 PRACTICAL SITUATION WHERE ASSAY 6072 04:20:16,800 --> 04:20:18,440 DEVELOPED ONE PLACE AND GETS 6073 04:20:18,440 --> 04:20:21,240 MOVED TO DIFFERENT PART OF THE 6074 04:20:21,240 --> 04:20:28,400 COMPANY OR MOVE TO CRO WHO TESTS 6075 04:20:28,400 --> 04:20:31,000 THE CON POUNDS. IF YOU RUN ASSAY 6076 04:20:31,000 --> 04:20:32,280 SUCCESSFULLY OR MAKE IMPORTANT 6077 04:20:32,280 --> 04:20:34,080 CHANGE TO REAGENT OR FOR MT. OF 6078 04:20:34,080 --> 04:20:37,000 ASSAY, THIS TEST FREE EXPERIMENT 6079 04:20:37,000 --> 04:20:39,560 I MENTIONED, ALL METRICS THAT I 6080 04:20:39,560 --> 04:20:41,280 REVIEWED, CAN BE VERY USEFUL FOR 6081 04:20:41,280 --> 04:20:48,520 THOSE PURPOSES. TO GET A BETTER 6082 04:20:48,520 --> 04:20:53,880 ESTIMATE OF MSR WE WANT TO 6083 04:20:53,880 --> 04:20:54,880 CAPTURE ENTERAND 6084 04:20:54,880 --> 04:20:55,760 INTRAVARYIBILITY. ONE WAY TO DO 6085 04:20:55,760 --> 04:20:57,240 THAT IS USE POSITIVE CONTROL 6086 04:20:57,240 --> 04:20:59,040 DATA OVER PERIOD OF TIME AND 6087 04:20:59,040 --> 04:21:02,640 CALCULATE WHAT WE CALL MOVING 6088 04:21:02,640 --> 04:21:06,200 MSR AND AFTER A WHILE USE 6089 04:21:06,200 --> 04:21:08,080 SCREENING DATA ITSELF TO 6090 04:21:08,080 --> 04:21:10,320 CALCULATE PERSPECTIVE MSR. SO 6091 04:21:10,320 --> 04:21:16,720 REVIEW WHAT MOVING MSR IS. MSR 6092 04:21:16,720 --> 04:21:19,880 AS IT WORKS YOU ARE CALCULATING 6093 04:21:19,880 --> 04:21:21,040 POSITIVE CONTROL MSR IN 6094 04:21:21,040 --> 04:21:23,160 INCREMENTS OF SIX RUNS USING THE 6095 04:21:23,160 --> 04:21:24,680 FIRST SIX RUNS CALCULATE THE 6096 04:21:24,680 --> 04:21:28,520 VARIABILITY OF POSITIVE CONTROL 6097 04:21:28,520 --> 04:21:32,080 IC 50 AND MSR TO SIX RUNS AND 6098 04:21:32,080 --> 04:21:34,800 RUNS TWO TO 7, THREE TO 8, SO 6099 04:21:34,800 --> 04:21:36,920 FORTH IN SLIDING INCREMENTS. 6100 04:21:36,920 --> 04:21:38,680 THAT IS WHY YOU HAVE THE MOVING 6101 04:21:38,680 --> 04:21:40,080 MSR AND YOU CAN BULK THE DATA 6102 04:21:40,080 --> 04:21:43,640 OVER TIME TO SEE HOW THE 6103 04:21:43,640 --> 04:21:44,640 VARIABILITY IS CHANGING OVER 6104 04:21:44,640 --> 04:21:46,400 TIME. HERE IS I WILL STRAIKS OF 6105 04:21:46,400 --> 04:21:48,560 THAT. THIS IS A POTENCY VALUE OF 6106 04:21:48,560 --> 04:21:51,560 DATA OF THE POSITIVE CONTROL, 6107 04:21:51,560 --> 04:21:55,480 PLOTTED OVER PERIOD OF TIME. WE 6108 04:21:55,480 --> 04:21:57,120 USE THE FIRST SIX RESULT TO 6109 04:21:57,120 --> 04:21:59,360 CALCULATE MSR VALUE. THAT IS 6110 04:21:59,360 --> 04:22:01,960 WHAT GIVES YOU THIS DOT HERE. 6111 04:22:01,960 --> 04:22:03,880 THEN RUNS TWO TO 7 GIVES YOU 6112 04:22:03,880 --> 04:22:05,920 THIS POINT HERE, RUNS THREE TO 8 6113 04:22:05,920 --> 04:22:09,520 GIVES YOU THE THIRD VALUE. SO 6114 04:22:09,520 --> 04:22:12,320 FORTH. SO EVERY DOT IS MSR 6115 04:22:12,320 --> 04:22:16,160 CALCULATED FOR SIX RUNS, SIX RUN 6116 04:22:16,160 --> 04:22:17,840 INCREMENTS OF ALL THESE POSITIVE 6117 04:22:17,840 --> 04:22:24,080 CONTROL IC 50 VALUES. SO REAL 6118 04:22:24,080 --> 04:22:26,240 TIME MONITOR HOW ASSAY BEHAVING 6119 04:22:26,240 --> 04:22:31,560 IN TERMS OF MSR THERE IS XL TOOL 6120 04:22:31,560 --> 04:22:34,120 IN THE AGM WEBSITE YOU PASTE THE 6121 04:22:34,120 --> 04:22:36,720 RUN DATE IN ONE COLUMN AND 6122 04:22:36,720 --> 04:22:38,920 CORRESPONDING IC 50 POSITIVE 6123 04:22:38,920 --> 04:22:42,880 CONTROL AND GIVES YOU THE MSR 6124 04:22:42,880 --> 04:22:44,040 VALUE, GRAPHS AND STATISTICS 6125 04:22:44,040 --> 04:22:48,440 NICELY. IT IS MUCH EASIER TO 6126 04:22:48,440 --> 04:22:51,320 FLAG ISSUES IN TERMS OF MSR THAN 6127 04:22:51,320 --> 04:22:52,600 POTENCY VALUE. HERE IS 6128 04:22:52,600 --> 04:22:54,920 ILLUSTRATION TO MAKE THE POINT. 6129 04:22:54,920 --> 04:22:56,160 HERE WE HAVE POTENCY DATA 6130 04:22:56,160 --> 04:22:58,720 PLOTTED OVER PERIOD OF TIME FROM 6131 04:22:58,720 --> 04:23:01,640 POSITIVE CONTROL FROM KINASE 6132 04:23:01,640 --> 04:23:02,760 ASSAY, LOOK AT CORRESPONDING 6133 04:23:02,760 --> 04:23:06,360 MOVING MSR GRAPH, FOR THE SAME 6134 04:23:06,360 --> 04:23:08,480 DATA, YOU CAN SEE SOME 6135 04:23:08,480 --> 04:23:10,360 VARIABILITY HERE BUT NOT SO 6136 04:23:10,360 --> 04:23:11,760 OBVIOUS THAT THERE IS REALLY BAD 6137 04:23:11,760 --> 04:23:14,320 OR ACCEPTABLE. LOOK AT 6138 04:23:14,320 --> 04:23:16,960 CORRESPONDING MSR FOR THE SAME 6139 04:23:16,960 --> 04:23:19,840 TIME WINDOW APRIL TO JULY, APRIL 6140 04:23:19,840 --> 04:23:21,360 TO JULY, IT IS MUCH WORSE IN 6141 04:23:21,360 --> 04:23:26,240 TERMS OF MSR, IT GOES UP FROM 6142 04:23:26,240 --> 04:23:28,720 THREE TO AROUND 7, 8, THEN COMES 6143 04:23:28,720 --> 04:23:31,920 BACK DOWN SO IN TERMS OF MOVING 6144 04:23:31,920 --> 04:23:35,640 MSR GIVES YOU MUCH BETTER FEEL 6145 04:23:35,640 --> 04:23:39,200 FOR HOW BAD HOW THE DATA IS THAN 6146 04:23:39,200 --> 04:23:41,160 SIMPLY LOOKING AT POTENCY VALUES 6147 04:23:41,160 --> 04:23:43,840 OVER TIME. THIS IS ANOTHER 6148 04:23:43,840 --> 04:23:46,920 EXAMPLE WITH THE SAME POINT, YOU 6149 04:23:46,920 --> 04:23:48,280 HAVE VARIABILITY TO START WITH 6150 04:23:48,280 --> 04:23:50,280 BUT IN TERMS OF MSR YOU GET A 6151 04:23:50,280 --> 04:23:52,840 APPRECIATION HOW BAD IT IS, SO 6152 04:23:52,840 --> 04:23:54,760 AROUND TWO, THREE, FOUR AND GOES 6153 04:23:54,760 --> 04:23:57,400 UP TO 12, 14, AFTER ISSUE IS 6154 04:23:57,400 --> 04:24:00,640 FIXED COMES DOWN TO AROUND 2. 6155 04:24:00,640 --> 04:24:02,640 ANOTHER ISSUE THAT COMES UP 6156 04:24:02,640 --> 04:24:05,560 LATER ON, WHERE YOU CAN SEE THE 6157 04:24:05,560 --> 04:24:07,760 DATA POTENCY VALUE GOING UP, 6158 04:24:07,760 --> 04:24:09,440 CORRESPONDING MSR IS ALSO VERY 6159 04:24:09,440 --> 04:24:13,440 HIGH. SO HELPS US TO MONITOR THE 6160 04:24:13,440 --> 04:24:17,200 QUALITY OF THE ASSAY OVER TIME 6161 04:24:17,200 --> 04:24:18,720 CONSISTENT POTENCY VALUES ARE 6162 04:24:18,720 --> 04:24:20,080 FOR POSITIVE CONTROL OVER TIME 6163 04:24:20,080 --> 04:24:21,680 AND HOW CONSISTENT MSR VALUES 6164 04:24:21,680 --> 04:24:22,880 ARE OVER TIME. AND IT IS 6165 04:24:22,880 --> 04:24:24,720 IMPORTANT TO DOCUMENT WHAT ARE 6166 04:24:24,720 --> 04:24:27,280 CHANGES HAPPENING IN THE ASSAY, 6167 04:24:27,280 --> 04:24:29,520 IMPORTANT TO DOCUMENT THAT AS 6168 04:24:29,520 --> 04:24:31,080 AND WHEN THEY HAPPEN. BUFFER 6169 04:24:31,080 --> 04:24:34,240 CHANGES OR REAGENT CHANGES OR 6170 04:24:34,240 --> 04:24:35,360 INCUBATION TIME OR OTHER 6171 04:24:35,360 --> 04:24:37,480 CONDITIONS ASSAY CHANGE. MAKE A 6172 04:24:37,480 --> 04:24:39,200 NOTE OF IT IN A TRACKING SHEET 6173 04:24:39,200 --> 04:24:41,240 AND LOOK AT CORRESPONDING MOVING 6174 04:24:41,240 --> 04:24:43,320 MSR VALUE, THE MOVING MSR GRAPH 6175 04:24:43,320 --> 04:24:44,720 YOU ARE TRACKING. IF IT LOOKS 6176 04:24:44,720 --> 04:24:46,760 LIKE THIS, AND MAKE A CHANGE IN 6177 04:24:46,760 --> 04:24:49,760 THESE NOT ONLY SEE MSR GO UP YOU 6178 04:24:49,760 --> 04:24:51,040 KNOW CHANGE IT WAS BEFORE CHANGE 6179 04:24:51,040 --> 04:24:52,320 REAGENT CHANGE THAT IS NOT A 6180 04:24:52,320 --> 04:24:55,200 GOOD THING. SO YOU HAVE TO GO 6181 04:24:55,200 --> 04:24:57,600 BACK AND SEE IF YOU CAN RESOLVE 6182 04:24:57,600 --> 04:25:04,440 THAT. THE THIRD MSR IS BASED ON 6183 04:25:04,440 --> 04:25:12,840 ALL DATA FROM SCREENING ASSAY. 6184 04:25:12,840 --> 04:25:13,640 AFTER FEW MONTHS GO BACK TO DATA 6185 04:25:13,640 --> 04:25:14,760 SET AND PULL OUT POSITIVE 6186 04:25:14,760 --> 04:25:16,440 CONTROL DATA AND ALL TEST 6187 04:25:16,440 --> 04:25:17,800 COMPOUNDS TESTED TWO OR MORE 6188 04:25:17,800 --> 04:25:21,440 TIMES. AGAIN ALL THE DATA IS 6189 04:25:21,440 --> 04:25:25,760 RUN THROUGH MIXED FX NOVA 6190 04:25:25,760 --> 04:25:27,240 NORMALIZING COMPOUND TO COMPOUND 6191 04:25:27,240 --> 04:25:28,400 DIFFERENCE, WHAT WE ARE LEFT 6192 04:25:28,400 --> 04:25:31,920 WITH IS VARIABILITY OF THE 6193 04:25:31,920 --> 04:25:35,120 SCREENING DATA AFTER NORMALIZING 6194 04:25:35,120 --> 04:25:40,320 COMPOUND DIFFERENCE. THAT IS 6195 04:25:40,320 --> 04:25:42,040 CAPTURED IN STUDENT VISUALS AND 6196 04:25:42,040 --> 04:25:46,840 PLOTTED OVER TIME. THIS IS THE 6197 04:25:46,840 --> 04:25:52,440 DATA ON SCREENING ASSAYS, 6198 04:25:52,440 --> 04:25:53,760 VARIABILITY OVER PERIOD OF TIME 6199 04:25:53,760 --> 04:25:55,040 FROM TESTING COMPOUND AND THIS 6200 04:25:55,040 --> 04:25:57,760 IS USED TO CALCULATE THE MSR. 6201 04:25:57,760 --> 04:26:01,240 THIS GIVES YOU GOOD FEEL OVERALL 6202 04:26:01,240 --> 04:26:03,280 VARIABILITY ASSAY AND TAKING ALL 6203 04:26:03,280 --> 04:26:05,920 THE DATA INTO ACCOUNT. SO 6204 04:26:05,920 --> 04:26:07,200 QUESTION THAT COMES UP WITH IS 6205 04:26:07,200 --> 04:26:09,320 IT REALLY NECESSARY TO USE ALL 6206 04:26:09,320 --> 04:26:13,080 THE DATA TO CALCULATE THE MSR. 6207 04:26:13,080 --> 04:26:15,520 OR ADEQUATE TO USE POSITIVE 6208 04:26:15,520 --> 04:26:17,960 CONTROL TO CALCULATE THE MSR? TO 6209 04:26:17,960 --> 04:26:19,720 ANSWER THIS QUESTION WE LOOK AT 6210 04:26:19,720 --> 04:26:21,720 TEN DIFFERENT ASSAYS, FIVE 6211 04:26:21,720 --> 04:26:25,720 KINASE ASSAYS AND CELL BASED 6212 04:26:25,720 --> 04:26:31,600 ASSAY AND BINDING ASSAY, MSR 6213 04:26:31,600 --> 04:26:34,520 FROM POSITIVE CONTROL THAT I 6214 04:26:34,520 --> 04:26:36,640 MENTION CALCULATEK SIX RUN 6215 04:26:36,640 --> 04:26:39,280 INCREMENTS YOU CALCULATE THE 6216 04:26:39,280 --> 04:26:40,720 POSITIVE CONTROL CELL OVER TIME 6217 04:26:40,720 --> 04:26:42,680 PERIOD TWO YEARS OR HOWEVER LOCK 6218 04:26:42,680 --> 04:26:44,920 SCREEN ASSAY IS. WE USE FOR 6219 04:26:44,920 --> 04:26:46,600 TESTING COMPOUNDS. CALCULATE THE 6220 04:26:46,600 --> 04:26:51,480 MSR FROM ALL THE POSITIVE 6221 04:26:51,480 --> 04:26:52,680 CONTROL RESULTS OVER PERIOD OF 6222 04:26:52,680 --> 04:26:53,600 TIME AND CALCULATE THE 6223 04:26:53,600 --> 04:26:56,600 CORRESPONDING MSRR. COMPARE THAT 6224 04:26:56,600 --> 04:26:58,960 TO RETROSPECTIVE MSR, AND TURNS 6225 04:26:58,960 --> 04:27:00,920 OUT IN ALL CASES WE HAVE SEEN 6226 04:27:00,920 --> 04:27:02,960 THEY TRACK WELL WITH THE MSR 6227 04:27:02,960 --> 04:27:05,320 CALCULATED USING ALL THE DATA 6228 04:27:05,320 --> 04:27:06,480 FROM THE TEST COMPOUNDS AS WELL 6229 04:27:06,480 --> 04:27:11,800 AS POSITIVE CONTROLS. SO THIS IS 6230 04:27:11,800 --> 04:27:12,640 FANTASTIC. THIS MEANS 6231 04:27:12,640 --> 04:27:13,480 VARIABILITY CALCULATE IN TERMS 6232 04:27:13,480 --> 04:27:15,400 OF POSITIVE CONTROL, IS PRETTY 6233 04:27:15,400 --> 04:27:16,560 GOOD REPRESENTATION OF THE 6234 04:27:16,560 --> 04:27:17,320 VARIABILITY YOU HAVE FROM ALL 6235 04:27:17,320 --> 04:27:19,600 THE DIFFERENT COMPOUNDS IN 6236 04:27:19,600 --> 04:27:21,040 SCREENING ASSAY. POSITIVE 6237 04:27:21,040 --> 04:27:25,480 CONTROL HAS TO BE THE ONE MOST 6238 04:27:25,480 --> 04:27:29,120 LOW IC 50 VALUE, LOW -- HIGH 6239 04:27:29,120 --> 04:27:33,720 LIPO TENT. YOU MAY WONDER DOES 6240 04:27:33,720 --> 04:27:39,600 IT PRESENT VARY -- HIGHLY 6241 04:27:39,600 --> 04:27:42,400 POTENT. YOU CAN SEE FROM THE 6242 04:27:42,400 --> 04:27:44,240 ASSAYS EVALUATED. THEY ARE IN 6243 04:27:44,240 --> 04:27:45,400 THE BALLPARK, RIGHT IN THE 6244 04:27:45,400 --> 04:27:48,640 MIDDLE MOST CASES. SO IN FACT 6245 04:27:48,640 --> 04:27:49,920 THIS MADE US FEEL SO GOOD ABOUT 6246 04:27:49,920 --> 04:27:55,400 IT THAT WE REDUCED THE FREQUENCY 6247 04:27:55,400 --> 04:27:57,800 RETROSPECTIVE MSR 6248 04:27:57,800 --> 04:27:58,480 A. CXFC DO IT EVERY FIVE MONTHS 6249 04:27:58,480 --> 04:28:00,120 AND MAKE SURE POSITIVE CONTROL 6250 04:28:00,120 --> 04:28:01,880 MSR IS CALCULATED AND KEEP TRACK 6251 04:28:01,880 --> 04:28:03,560 OF MOVING MSR OVER PERIOD OF 6252 04:28:03,560 --> 04:28:07,360 TIME WHEN WE GENERATE DATA. THIS 6253 04:28:07,360 --> 04:28:08,680 COME SEPTEMBER OF MSR IS 6254 04:28:08,680 --> 04:28:10,440 EXTENNED TO SO FAR WE TALKED 6255 04:28:10,440 --> 04:28:13,080 ABOUT COMPARING HOW RELIABLE YOU 6256 04:28:13,080 --> 04:28:16,800 CAN DIFFERENTIATE POTENTIAL 6257 04:28:16,800 --> 04:28:18,080 CELLS ANY PAIR OF COMPOUNDS IN 6258 04:28:18,080 --> 04:28:21,160 THE SCREENING ASSAY. THE OTHER 6259 04:28:21,160 --> 04:28:27,240 IMPORTANT METRIC IS SELECTIVITY 6260 04:28:27,240 --> 04:28:28,600 RATIO, SO YOUR NOT ONLY 6261 04:28:28,600 --> 04:28:30,800 INTERESTED IN ACTIVITY OF 6262 04:28:30,800 --> 04:28:32,240 AGAINST PRIMARY TARGET BUT MAKE 6263 04:28:32,240 --> 04:28:35,240 SURE COMPOUNDS DON'T BIND OR NOT 6264 04:28:35,240 --> 04:28:38,160 ACTIVE AGAINST OFF TARGET SO 6265 04:28:38,160 --> 04:28:39,560 SELECT RASH YES REFLECTS THE 6266 04:28:39,560 --> 04:28:43,160 RATIO OF POTENCY TO PRIMARY 6267 04:28:43,160 --> 04:28:44,680 TARGET VERSUS POTENCY AGAINST 6268 04:28:44,680 --> 04:28:52,320 THE OFF TARGET. IF YOU WANT 6269 04:28:52,320 --> 04:28:54,360 SELECTIVITY RATIO FOR ALL 6270 04:28:54,360 --> 04:28:57,160 COMPOUNDS SELECTED FROM OFF 6271 04:28:57,160 --> 04:28:58,440 TARGET, BUT THERE WE HAVE EVEN 6272 04:28:58,440 --> 04:29:00,280 MORE VARIABILITY. WE HAVE 6273 04:29:00,280 --> 04:29:01,960 VARIABILITY PRIMARY ASSAY, 6274 04:29:01,960 --> 04:29:03,240 PRIMARY TARGET ASSAY AND 6275 04:29:03,240 --> 04:29:04,200 VARIABILITY OF THE OFF TARGET 6276 04:29:04,200 --> 04:29:09,640 ASSAY. PROPAGATION OF ERROR AS 6277 04:29:09,640 --> 04:29:12,160 YOU ADD MORE DATA TO THE 6278 04:29:12,160 --> 04:29:13,760 DIVISION. SO THERE IS A SIMPLE 6279 04:29:13,760 --> 04:29:16,680 EXTENSION OF MSR IN SELECTIVITY 6280 04:29:16,680 --> 04:29:20,800 RATIO APPLICATION. BECAUSE MSSR 6281 04:29:20,800 --> 04:29:22,880 MSR SR STANDS FOR MINIMUM 6282 04:29:22,880 --> 04:29:24,160 SIGNIFICANT SELECTIVITY RATIO 6283 04:29:24,160 --> 04:29:28,000 AND MSR SR IS MINIMUM 6284 04:29:28,000 --> 04:29:30,200 SIGNIFICANT RATIO OF SELECTIVITY 6285 04:29:30,200 --> 04:29:32,040 RATIOS, VERY EASY TO CALCULATE. 6286 04:29:32,040 --> 04:29:34,480 USING JUST THE MSR YOU CALCULATE 6287 04:29:34,480 --> 04:29:38,920 FROM THE PRIMARY ASSAY, THE OFF 6288 04:29:38,920 --> 04:29:39,840 TARGET YOU CAN CALCULATE 6289 04:29:39,840 --> 04:29:46,120 METRICS. HERE IS MSSR THIS IS 6290 04:29:46,120 --> 04:29:48,720 FOR ASSESS WHETHER COMPOUND IS 6291 04:29:48,720 --> 04:29:52,160 DIFFERENT ON TWO TARGETS PRIMARY 6292 04:29:52,160 --> 04:29:57,720 TARGET OFF TARGET MSSR PRESENTS 6293 04:29:57,720 --> 04:29:59,640 SMALL CHANGE FROM POTENCY TO 6294 04:29:59,640 --> 04:30:01,680 PRIMARY TO PROTEN SHY TO OFF 6295 04:30:01,680 --> 04:30:04,360 TARGETOR TARGET. SO THIS CALLS 6296 04:30:04,360 --> 04:30:06,840 FOR PLUGGENING MSR VALUE FOR THE 6297 04:30:06,840 --> 04:30:10,040 ASSAY, ASSAY MSR EQUATION, THAT 6298 04:30:10,040 --> 04:30:15,320 GIVES YOU THE MSSR VALUE. MSSR 6299 04:30:15,320 --> 04:30:17,320 PRIMARY IS 2.5 AND FOR 6300 04:30:17,320 --> 04:30:19,680 SELECTIVITY ASSAY IS THREE. PLUG 6301 04:30:19,680 --> 04:30:24,120 THOSE INTO THE FORMULA, MSSR IS 6302 04:30:24,120 --> 04:30:25,960 2.75 SO HOW WE USE IN PRACTICE. 6303 04:30:25,960 --> 04:30:29,000 SUPPOSE CRITERIA IS 100 FOLD YOU 6304 04:30:29,000 --> 04:30:30,600 WANT COMPOUNDS 100 FOR SELECTIVE 6305 04:30:30,600 --> 04:30:33,440 TO THE PRIMARY TARGET AND 6306 04:30:33,440 --> 04:30:34,840 DURATION OFF TARGET. WE TALKED 6307 04:30:34,840 --> 04:30:38,320 ABOUT VARIABILITY. QUITE A LOT. 6308 04:30:38,320 --> 04:30:40,280 BOTH ASSAYS NOW AND WE TAKE 6309 04:30:40,280 --> 04:30:41,360 VARIABILITY IN BOTH ASSAYS INTO 6310 04:30:41,360 --> 04:30:44,560 ACCOUNT. THE MSSR IS 2.75, WHAT 6311 04:30:44,560 --> 04:30:46,920 DOES IT MEAN IN TERMS OF HUNDRED 6312 04:30:46,920 --> 04:30:48,880 FOLD CRITERIA? MEANS THAT I NEED 6313 04:30:48,880 --> 04:30:52,760 TO SEE AT LEAST 275 FOLD 6314 04:30:52,760 --> 04:30:55,760 SELECTIVITY TO BE 95% CONFIDENT 6315 04:30:55,760 --> 04:30:56,520 THAT I HAVE 100 FOLD 6316 04:30:56,520 --> 04:31:01,400 SELECTIVITY. IF THAT IS TOO MUCH 6317 04:31:01,400 --> 04:31:04,600 TEST COMPOUNDS TWICE AND BRING 6318 04:31:04,600 --> 04:31:07,320 MSSR DOWN TO 2.2 OR 2. THAT 6319 04:31:07,320 --> 04:31:10,240 MEANS IF CRITERIA IS 100 FOLD 6320 04:31:10,240 --> 04:31:12,000 SELECTIVITY YOU NEED AT LEAST 6321 04:31:12,000 --> 04:31:14,920 200 FOLD SELECTIVITY TO BE 95% 6322 04:31:14,920 --> 04:31:16,080 CONFIDENT YOU HAVE 100 FOLD 6323 04:31:16,080 --> 04:31:20,320 SELECTIVITY. OTHER THINGS ALSO 6324 04:31:20,320 --> 04:31:21,960 WHAT IF I HAVE FIVE FOLD 6325 04:31:21,960 --> 04:31:23,680 SELECTIVITY? DOES THAT MEAN I 6326 04:31:23,680 --> 04:31:26,640 SHOULD MOVE COMPOUND FORWARD? 6327 04:31:26,640 --> 04:31:28,240 THESE ARE BASED ON 95% 6328 04:31:28,240 --> 04:31:30,400 CONFIDENCE. SO WE CAN REVERSE 6329 04:31:30,400 --> 04:31:33,520 ENGINEER THIS AND CALCULATE WHAT 6330 04:31:33,520 --> 04:31:34,760 LITTLE CONFIDENCE CAN YOU HAVE 6331 04:31:34,760 --> 04:31:37,840 WITH COMPOUND THAT HAS 150 FOLD 6332 04:31:37,840 --> 04:31:40,760 SELECTIVITY. IT DOESN'T HAVE THE 6333 04:31:40,760 --> 04:31:43,320 275 FOLD SELECTIVITY BUT 150 6334 04:31:43,320 --> 04:31:45,400 FOLD SELECTIVITY, HOW MANY 6335 04:31:45,400 --> 04:31:49,320 CONFIDENCE DO I HAVE? I DON'T 6336 04:31:49,320 --> 04:31:51,680 HAVE 95% CONFIDENCE BUT DO I 6337 04:31:51,680 --> 04:31:54,960 HAVE 75%? THAT IS COVERED IN ONE 6338 04:31:54,960 --> 04:31:56,520 OF THE PAPERS I CITED IN THE 6339 04:31:56,520 --> 04:31:58,960 FIRST SLIDE, THE THIRD CITATION 6340 04:31:58,960 --> 04:32:00,400 OF THE PRESENTATION. THAT 6341 04:32:00,400 --> 04:32:01,920 PUBLICATION WE GO OVER THAT IN 6342 04:32:01,920 --> 04:32:08,600 DETAIL HOW TO CALCULATE 6343 04:32:08,600 --> 04:32:12,840 VARIABILITY OF COMPOUND BEING 6344 04:32:12,840 --> 04:32:14,240 MORE HOE SELECTIVITY OF RATIO 6345 04:32:14,240 --> 04:32:16,040 SIGNIFICANTLY HIGHER THAN THE 6346 04:32:16,040 --> 04:32:17,360 HUNDRED FOLD SELECTIVITY AS IN 6347 04:32:17,360 --> 04:32:21,480 THIS CASE. SO WE DONE HAVE TO 6348 04:32:21,480 --> 04:32:23,720 BLINDLY APPLY CONCEPTS IN TERMS 6349 04:32:23,720 --> 04:32:27,120 OF 95% CONFIDENCE YOU CAN 6350 04:32:27,120 --> 04:32:29,800 CALCULATE THE CONFIDENCE LEVEL 6351 04:32:29,800 --> 04:32:31,400 ITSELF FOR A GIVEN COMPOUND IF 6352 04:32:31,400 --> 04:32:36,720 IT DOESN'T MEET MSSR CRITERIA. 6353 04:32:36,720 --> 04:32:39,160 EXTENDING FURTHER NOW IF YOUR 6354 04:32:39,160 --> 04:32:40,000 HONOR PROGRAM CALLS FOR 6355 04:32:40,000 --> 04:32:41,360 COMPOUNDS HAVING BETTER 6356 04:32:41,360 --> 04:32:44,360 SELECTIVITY THAN POSITIVE 6357 04:32:44,360 --> 04:32:46,440 CONTROL, SUPPOSE POSITIVE 6358 04:32:46,440 --> 04:32:48,120 CONTROL HAS 50 FOLD SELECTIVITY 6359 04:32:48,120 --> 04:32:49,800 AGAINST OFF TARGE TARGET. WE WT 6360 04:32:49,800 --> 04:32:52,440 TEST COMPOUNDS MOVE FORWARD TO 6361 04:32:52,440 --> 04:32:53,880 BEAT POSITIVE CONTROL HAVING 6362 04:32:53,880 --> 04:32:55,880 MORE THAN 50 FOLD SELECTIVITY 6363 04:32:55,880 --> 04:33:02,160 RATIO THIS HAS. BETWEEN TEST 6364 04:33:02,160 --> 04:33:03,600 COMPOUNDS OR POSITIVE CONTROL 6365 04:33:03,600 --> 04:33:04,920 AND ALSO TEST COMPOUNDS. FOR 6366 04:33:04,920 --> 04:33:09,200 THAT WE NEED MSR SR AND WE PLUG 6367 04:33:09,200 --> 04:33:11,280 IN THOSE TWO VALUES AS IN THE 6368 04:33:11,280 --> 04:33:12,680 PREVIOUS SLIDE WITH A SLIGHTLY 6369 04:33:12,680 --> 04:33:17,440 DIFFERENT FORMULA. PURPOSE TO 6370 04:33:17,440 --> 04:33:19,440 COMPARE SELECTIVITY RATIOS. FOR 6371 04:33:19,440 --> 04:33:21,640 EXAMPLE, MSR PRIMARY ASSAY IS 6372 04:33:21,640 --> 04:33:24,840 TWO, OFF TARGET MSR IS THREE. 6373 04:33:24,840 --> 04:33:27,680 MSR IS 3.67. WE WANT COMPOUNDS 6374 04:33:27,680 --> 04:33:30,800 TO BE AT LEAST 50 FOLD SELECTED 6375 04:33:30,800 --> 04:33:33,920 THAT MEANS AT LEAST 180 FOLD 6376 04:33:33,920 --> 04:33:40,920 SELECTIVITY TO BE 95% CONFIDENCE 6377 04:33:40,920 --> 04:33:42,160 FOR 50 FOALED SELECTION. THIS 6378 04:33:42,160 --> 04:33:43,680 SEEMS LIKE A BIG NUMBER BUT IF 6379 04:33:43,680 --> 04:33:44,640 YOU TEST COMPOUND MORE THAN ONCE 6380 04:33:44,640 --> 04:33:47,520 WE CAN BRING THE VALUE DOWN AND 6381 04:33:47,520 --> 04:33:49,480 CALCULATE THE CORRESPONDING MSR 6382 04:33:49,480 --> 04:33:58,520 SR. WHEN YOU HAVE TWO 6383 04:33:58,520 --> 04:34:00,480 COMPOUNDS, ONE SELECTIVITY RATIO 6384 04:34:00,480 --> 04:34:02,960 OF 17, ANOTHER COMPOUND 6385 04:34:02,960 --> 04:34:09,520 SELECTIVITY RATIO OF 73. SO THIS 6386 04:34:09,520 --> 04:34:11,800 SECOND COMPOUND IS FOUR FOLD 6387 04:34:11,800 --> 04:34:13,080 MORE SELECTIVE THAN THE FIRST. 6388 04:34:13,080 --> 04:34:15,640 IF YOU TESTED ONLY ONCE, THE MSR 6389 04:34:15,640 --> 04:34:18,160 SR IS 5.6 SO NOT GOOD ENOUGH. 6390 04:34:18,160 --> 04:34:21,040 BUT IF YOU TESTED THESE 6391 04:34:21,040 --> 04:34:25,200 COMPOUNDS TWICE 5.6 COMES DOWN 6392 04:34:25,200 --> 04:34:26,640 TO 3.3. SO WE HAVE THE 6393 04:34:26,640 --> 04:34:27,960 CONFIDENCE THAT THESE COMPOUNDS 6394 04:34:27,960 --> 04:34:29,520 IS MUCH MORE SELECTIVE THAN THE 6395 04:34:29,520 --> 04:34:35,080 FIRST COMPOUND. SO THIS HELPS 6396 04:34:35,080 --> 04:34:36,160 DECIDE HOW MANY TIMES TO TEST 6397 04:34:36,160 --> 04:34:39,400 THE COMPOUNDS AND HOW WELL 6398 04:34:39,400 --> 04:34:44,360 DIFFERENTIATE THESE COMPOUNDS. 6399 04:34:44,360 --> 04:34:46,720 IN SUMMARY YOU HEARD Z PRIME IN 6400 04:34:46,720 --> 04:34:48,280 DETAIL OVER THE PREVIOUS 6401 04:34:48,280 --> 04:34:49,960 PRESENTATION AN HEARD HOW TO 6402 04:34:49,960 --> 04:34:51,800 EXTEND THE Z PRIME FACTOR TO 6403 04:34:51,800 --> 04:34:56,920 CONTEXT OF POTENCY DATA MAKING 6404 04:34:56,920 --> 04:34:58,280 IMPORTANT DECISIONS IN TERMS OF 6405 04:34:58,280 --> 04:34:59,480 ONE COMPOUND BETTER THAN THE 6406 04:34:59,480 --> 04:35:01,360 OTHER IN TERMS OF POTENCY VALUES 6407 04:35:01,360 --> 04:35:03,480 OR SELECTIVITY RATIOS WE HAVE 6408 04:35:03,480 --> 04:35:04,920 DIFFERENT TYPE OF M SR YOU CAN 6409 04:35:04,920 --> 04:35:06,880 CALCULATE. WE ALSO TALKED ABOUT 6410 04:35:06,880 --> 04:35:08,320 HOW WE CAN MONITOR THE QUALITY 6411 04:35:08,320 --> 04:35:10,880 OF ASSAY IN REAL TIME USING 6412 04:35:10,880 --> 04:35:12,120 POSITIVE CONTROL DATA AND NOT 6413 04:35:12,120 --> 04:35:14,560 JUST LOOKING AT POSITIVE POTENCY 6414 04:35:14,560 --> 04:35:18,880 OVER TIME BUT LOOKING AT THE 6415 04:35:18,880 --> 04:35:20,200 VARIABILITY MILLION TERMS OF MSR 6416 04:35:20,200 --> 04:35:21,240 POSITIVE CONTROL OVER TIME 6417 04:35:21,240 --> 04:35:23,560 MAKING OBJECTIVE POSITION ON IS 6418 04:35:23,560 --> 04:35:28,400 THERE CHANGE IN REAGENT OR ASSAY 6419 04:35:28,400 --> 04:35:29,800 MEANINGFUL, DOES IT REDUCE 6420 04:35:29,800 --> 04:35:31,000 QUALITY OF ASSAY SO IT GETS ALL 6421 04:35:31,000 --> 04:35:32,680 THAT AND WE TALKED ABOUT HOW TO 6422 04:35:32,680 --> 04:35:34,800 EXTEND THE CONTEMPT IN TERMS OF 6423 04:35:34,800 --> 04:35:37,040 RATIO AS WELL. SO WITH THAT I'LL 6424 04:35:37,040 --> 04:35:47,520 STOP. THANK YOU VERY MUCH. 6425 04:35:49,560 --> 04:35:52,640 >>THAT WAS A FANTASTIC TALK. 6426 04:35:52,640 --> 04:35:55,040 THE Z PRIME FACTOR EQUATION, 6427 04:35:55,040 --> 04:35:58,080 DESCRIBED THE SEPARATION BETWEEN 6428 04:35:58,080 --> 04:36:00,280 HIGH AND LOW CONTROLS. IT IS TC 6429 04:36:00,280 --> 04:36:03,440 SHOWED US WHEN WE LOOK AT THE 6430 04:36:03,440 --> 04:36:05,560 EQUATION, IT IS ANATOMY TELLS 6431 04:36:05,560 --> 04:36:06,920 EXACTLY HOW TO DEVELOP A HIGH 6432 04:36:06,920 --> 04:36:09,480 QUALITY ASSAY TO IDENTIFY HITS, 6433 04:36:09,480 --> 04:36:11,160 RIGHT? WE WANT TO MAXIMIZE THE 6434 04:36:11,160 --> 04:36:12,880 SIGNAL TO BACKGROUND, WE WANT TO 6435 04:36:12,880 --> 04:36:17,000 MINIMIZE THE VARIABILITY. I KNOW 6436 04:36:17,000 --> 04:36:18,520 YOU HAVE HAD EXPERIENCES OVER 6437 04:36:18,520 --> 04:36:20,440 THE YEARS WORKING WITH ASSAY 6438 04:36:20,440 --> 04:36:23,520 BIOLOGISTS, ON THE MSR WHICH 6439 04:36:23,520 --> 04:36:27,760 HELPS PEOPLE DEVELOP A GOOD 6440 04:36:27,760 --> 04:36:32,160 ASSAY COMPARE POTENCIES. THE 6441 04:36:32,160 --> 04:36:34,560 MSR UNLIKE Z FACTOR TELLS YOU 6442 04:36:34,560 --> 04:36:35,360 SEPARATION BETWEEN HIGH AN LOW 6443 04:36:35,360 --> 04:36:39,000 CONTROLS, THE MSR TELLS YOU 6444 04:36:39,000 --> 04:36:40,880 VARIABILITY OF POTENCY. BUT IT 6445 04:36:40,880 --> 04:36:43,960 IS LESS CLEAR FROM EQUATION WHAT 6446 04:36:43,960 --> 04:36:46,360 WE CAN DO IS AS BIOLOGIST TO 6447 04:36:46,360 --> 04:36:52,800 MAKE THE BEST POSSIBLE ASSAYS TO 6448 04:36:52,800 --> 04:36:55,040 OPTIMIZE POTENCY MEASUREMENTS OR 6449 04:36:55,040 --> 04:36:56,320 REDUCE VARIABILITY OF 6450 04:36:56,320 --> 04:36:58,920 MEASUREMENTS. WERE YOU ABLE TO 6451 04:36:58,920 --> 04:37:00,360 LEARN ANYTHING FROM ASSAY 6452 04:37:00,360 --> 04:37:03,000 BIOLOGIST COLLEAGUES ABOUT 6453 04:37:03,000 --> 04:37:04,200 THINGS THAT HAVEN'T THOSE ASSAYS 6454 04:37:04,200 --> 04:37:07,680 TO REDUCE MSR? 6455 04:37:07,680 --> 04:37:12,200 >>GREAT QUESTION. WE TALKED Z 6456 04:37:12,200 --> 04:37:14,920 PRIME OF .7 DOESN'T MEAN MSR OR 6457 04:37:14,920 --> 04:37:16,920 2 OR VERY GOOD MSR VALUE SO IF 6458 04:37:16,920 --> 04:37:18,520 YOU OPTIMIZE THE ASSAY IN TERMS 6459 04:37:18,520 --> 04:37:24,920 OF JUST Z PRIME DOES IT 6460 04:37:24,920 --> 04:37:26,680 GUARANTEE GOOD ASAYS AND HOW DO 6461 04:37:26,680 --> 04:37:27,920 YOU WORK AROUND THAT AND 6462 04:37:27,920 --> 04:37:29,400 OPTIMIZE ASSAY TO MAKE SURE YOU 6463 04:37:29,400 --> 04:37:30,920 HAVE GOOD MSR. THAT IS WHAT YOU 6464 04:37:30,920 --> 04:37:35,920 ARE ASKING. SO ACTUALLY HAD 6465 04:37:35,920 --> 04:37:36,800 SITUATION A NUMBER OF TIMES 6466 04:37:36,800 --> 04:37:39,160 WHERE THE MSR WAS BAD, EXAMPLE 6467 04:37:39,160 --> 04:37:41,200 WAS BRIEFLY MENTIONED ABOUT 6468 04:37:41,200 --> 04:37:44,000 EARLIER FOR CELL BASED ASSAY, I 6469 04:37:44,000 --> 04:37:45,360 CAN'T TELL THEM SCREEN TWO OR 6470 04:37:45,360 --> 04:37:47,280 THREE TIMES FOR LOW THROUGH PUT 6471 04:37:47,280 --> 04:37:49,280 ASSAY, YOU WANT TO REOPTIMIZE IT 6472 04:37:49,280 --> 04:37:51,360 YOU HAVE TO A MULTI-FACTOR DOE 6473 04:37:51,360 --> 04:37:53,120 APPROACH TO OPTIMIZE THE ASSAY. 6474 04:37:53,120 --> 04:37:55,880 THE CRITERIA THAT WE USE THERE, 6475 04:37:55,880 --> 04:37:57,680 THE REASON FIRST OPT MICE 6476 04:37:57,680 --> 04:38:02,120 BECAUSE MSR WAS AROUND 6 OR 7. 6477 04:38:02,120 --> 04:38:04,320 SO THEY WANT TO BRING DOWN TO 6478 04:38:04,320 --> 04:38:08,760 LESS THAN THREE. WHAT WE DID WAS 6479 04:38:08,760 --> 04:38:12,200 SET UP THE DESIGN IN SUCH A WAY 6480 04:38:12,200 --> 04:38:15,400 DOE IN A WAY TO GENERATE DATA 6481 04:38:15,400 --> 04:38:19,160 NOT JUST LOW AND HIGH BUT TWO 6482 04:38:19,160 --> 04:38:25,200 OTHER CONCENTRATIONS IN BETWEEN 6483 04:38:25,200 --> 04:38:26,720 BACKGROUND, ONE LEVEL OF LOW, 6484 04:38:26,720 --> 04:38:29,400 AND MID LEVEL, AND ANOTHER LEVEL 6485 04:38:29,400 --> 04:38:33,360 BETWEEN MID AND HIGH. THEN HIGH 6486 04:38:33,360 --> 04:38:35,640 THEN GENERATE REPLICATES AT EACH 6487 04:38:35,640 --> 04:38:40,440 OF THOSE THEN WE CALCULATE THE 6488 04:38:40,440 --> 04:38:41,560 VARIABILITY AROUND EACH PAIR 6489 04:38:41,560 --> 04:38:43,160 BACKGROUND TO LOW, BACKGROUND TO 6490 04:38:43,160 --> 04:38:45,160 MID, BACKGROUND TO MODERATE AND 6491 04:38:45,160 --> 04:38:46,920 BACKGROUND TO HIGH. AND WE USED 6492 04:38:46,920 --> 04:38:51,480 ALL THESE MEASURES IN THE DOE 6493 04:38:51,480 --> 04:38:53,200 ANALYSIS TO FIGURE OUT THE BEST 6494 04:38:53,200 --> 04:38:57,000 ASSAY CONDITIONS THAT WILL GIVE 6495 04:38:57,000 --> 04:38:58,080 YOU GOOD SEPARATION ACROSS ALL 6496 04:38:58,080 --> 04:39:01,040 THE WINDOWS THAT WE HAVE. FROM 6497 04:39:01,040 --> 04:39:04,840 DIFFERENT CONCENTRATIONS. THAT 6498 04:39:04,840 --> 04:39:08,640 WILL GET YOU CLOSER TO HAVING 6499 04:39:08,640 --> 04:39:12,040 REASONABLY GOOD DATA ACROSS THE 6500 04:39:12,040 --> 04:39:13,280 ENTIRE RANGE CONCENTRATION 6501 04:39:13,280 --> 04:39:15,960 RESPONSE CURVE. WE DON'T KNOW WE 6502 04:39:15,960 --> 04:39:17,760 HAVE ONE COMPOUND IC 50 COULD BE 6503 04:39:17,760 --> 04:39:19,360 DOWN HERE, ANOTHER COMPOUND IC 6504 04:39:19,360 --> 04:39:22,800 50 COULD BE UP HERE. WE WANT TO 6505 04:39:22,800 --> 04:39:27,000 MAKE SURE THE ENTIRETY ENTIRE 6506 04:39:27,000 --> 04:39:28,600 AGENT COURSE RESPONSE -- DOSE 6507 04:39:28,600 --> 04:39:29,600 RESPONSE CURVE WE HAVE 6508 04:39:29,600 --> 04:39:30,840 REASONABLY GOOD DATA WITH LOW 6509 04:39:30,840 --> 04:39:33,560 VARIABILITY AND ROBUST SIGNAL. 6510 04:39:33,560 --> 04:39:34,400 THIS IS A WAY THE 6511 04:39:34,400 --> 04:39:38,200 >>ABOUT IT. TO OPTIMIZE 6512 04:39:38,200 --> 04:39:39,760 QUALITY AT DIFFERENT SECTIONS OF 6513 04:39:39,760 --> 04:39:42,880 THE POSITIVE CONTROL 6514 04:39:42,880 --> 04:39:43,800 CONCENTRATION RESPONSE CURVE. 6515 04:39:43,800 --> 04:39:46,920 THAT WAY YOU CAN HOPE THAT 6516 04:39:46,920 --> 04:39:48,800 EVENTUALLY BEST ASSAY CONDITION 6517 04:39:48,800 --> 04:39:51,920 YOU COME UP WITH THAT OPTIMIZES 6518 04:39:51,920 --> 04:39:56,360 IT WILL GIVE YOU ROBUST IC 50 6519 04:39:56,360 --> 04:39:57,800 DATA. OBVIOUSLY IT WAS 6520 04:39:57,800 --> 04:39:59,600 SUCCESSFUL WE BROUGHT DOWN MSR 6521 04:39:59,600 --> 04:40:01,280 IN ONE CASE FROM CIRCUMSTANCES 6522 04:40:01,280 --> 04:40:04,480 OR SEVEN DOWN TO THREE OR FOUR. 6523 04:40:04,480 --> 04:40:06,000 OBVIOUSLY WE CAN ONLY DO BETTER. 6524 04:40:06,000 --> 04:40:12,480 CAN'T DO WORSE WORSE BUT THIS IY 6525 04:40:12,480 --> 04:40:13,960 TO GO AROUND CHALLENGE OF 6526 04:40:13,960 --> 04:40:17,280 RELIABLE IC 50 VALUES BUT WORK 6527 04:40:17,280 --> 04:40:19,080 DIFFERENT SEGMENTS OF THE 6528 04:40:19,080 --> 04:40:20,320 CONCENTRATION RESPONSE CURVE NOT 6529 04:40:20,320 --> 04:40:23,560 JUST HIGH AND LOW. 6530 04:40:23,560 --> 04:40:25,160 >>I THINK THIS MAYBE A GOOD 6531 04:40:25,160 --> 04:40:28,280 TIME TO ASK ABOUT BEST PRACTICE 6532 04:40:28,280 --> 04:40:35,280 FOR IC 50. I SEE SOME ISSUES, 6533 04:40:35,280 --> 04:40:37,240 SOMETIMES THE CURVE REACH 60%, 6534 04:40:37,240 --> 04:40:42,280 HOW DO YOU FIX THE BEST IC 50? I 6535 04:40:42,280 --> 04:40:46,520 HEARD SOME USE 100% MANDATORY TO 6536 04:40:46,520 --> 04:40:49,520 100 SOME USE AOC. SO THAT IS MY 6537 04:40:49,520 --> 04:40:51,320 FIRST QUESTION. MY SECOND 6538 04:40:51,320 --> 04:40:52,960 QUESTION, SOMETIMES ALSO THE 6539 04:40:52,960 --> 04:40:57,560 CURVE SEEMS TO DRIFT AND MINUS 6540 04:40:57,560 --> 04:40:59,120 20, IN THAT CASE WHAT WILL BE 6541 04:40:59,120 --> 04:41:00,840 THE BEST WAY? 6542 04:41:00,840 --> 04:41:02,240 >>PROBABLY ASKING ABOUT THOSE 6543 04:41:02,240 --> 04:41:04,120 ASSAYS WHERE YOU DON'T HAVE A 6544 04:41:04,120 --> 04:41:07,640 CLEAR IC 50 WHERE YOU DON'T HAVE 6545 04:41:07,640 --> 04:41:11,400 A CLEAR MAX. WE HAVE SOME 6546 04:41:11,400 --> 04:41:13,000 PERCENT EFFICACY. 6547 04:41:13,000 --> 04:41:15,600 >>SAY WE DO HAVE A GOOD 6548 04:41:15,600 --> 04:41:18,080 POSITIVE CONTROL TO GET 100%, 6549 04:41:18,080 --> 04:41:19,680 BUT FOR CERTAIN COMPOUNDS 6550 04:41:19,680 --> 04:41:20,000 DOESN'T GET -- 6551 04:41:20,000 --> 04:41:22,960 >>SO SOME COMPONENTS YOU GET TO 6552 04:41:22,960 --> 04:41:27,800 70%, SOME 90, SOME 60. SO YOU 6553 04:41:27,800 --> 04:41:29,600 ASK HOW DO WE DEAL WITH THOSE 6554 04:41:29,600 --> 04:41:31,960 KINDS OF DATA AND HOW DO YOU 6555 04:41:31,960 --> 04:41:34,480 DEFINE THE EC 50 IN THOSE CASES? 6556 04:41:34,480 --> 04:41:45,000 >>COMPARE THEM.ONE IS TO HAVE 6557 04:41:45,880 --> 04:41:47,080 A THRESHOLD FOR ALL THE 6558 04:41:47,080 --> 04:41:50,080 COMPOUNDS, SO DEFINING THE EC 50 6559 04:41:50,080 --> 04:41:51,800 IN REALS TO THE MAXIMUM OBSERVED 6560 04:41:51,800 --> 04:41:53,040 -- IN RELATION TO THE MAXIMUM 6561 04:41:53,040 --> 04:41:54,560 OBSERVED ACTIVITY FOR THAT 6562 04:41:54,560 --> 04:41:57,520 COMPOUND BUT DEFINING IT AS 6563 04:41:57,520 --> 04:42:00,880 ABSOLUTE, ABSOLUTE 50% SCALE. SO 6564 04:42:00,880 --> 04:42:03,760 GIVEN COMPOUNDS MAXIMUM IS 70, 6565 04:42:03,760 --> 04:42:06,840 YOU CALCULATE CONCENTRATION OF 6566 04:42:06,840 --> 04:42:10,280 50% RAW 50% ACTIVITY, 50% OF 70 6567 04:42:10,280 --> 04:42:17,440 BUT 50% PERIOD. SO CALCULATE 6568 04:42:17,440 --> 04:42:20,520 50% A I LONG WITH AOCC. SOME MAY 6569 04:42:20,520 --> 04:42:22,480 NOT AGREE WITH IT BUT IF YOU 6570 04:42:22,480 --> 04:42:24,600 COMBINE WITH AOC, WE FIND IT TO 6571 04:42:24,600 --> 04:42:27,600 BE PRETTY GOOD. SO YOU CAN 6572 04:42:27,600 --> 04:42:30,520 CALCULATE END OF THE CURVE FOR 6573 04:42:30,520 --> 04:42:31,840 THE ENTIRE CONCENTRATION 6574 04:42:31,840 --> 04:42:34,600 RESPONSE CURVE USE AUC WITH 6575 04:42:34,600 --> 04:42:43,000 ABSOLUTE EC 50 TO COMPOUNDS. 6576 04:42:43,000 --> 04:42:45,680 (INAUDIBLE) ANOTHER WAY IS TO 6577 04:42:45,680 --> 04:42:49,800 CALCULATE RELATION TO MAXIMUM 6578 04:42:49,800 --> 04:42:54,520 OBSERVED ACTIVITY. AND THEN GO 6579 04:42:54,520 --> 04:43:04,960 ALONGTHAT IS THE POTENCY. 6580 04:44:13,720 --> 04:44:15,800 THE EFFICACY IS THE PLATEAU. IF 6581 04:44:15,800 --> 04:44:18,320 IT IS LIKE 50% PARTIAL AGONIST. 6582 04:44:18,320 --> 04:44:20,480 YOU CAN ALSO SUPER AGONIST WHICH 6583 04:44:20,480 --> 04:44:22,920 GOES ABOVE 100%. SO THAT IS A 6584 04:44:22,920 --> 04:44:27,880 PHARMACOLOGIC -- THE CASES WHERE 6585 04:44:27,880 --> 04:44:29,480 MATHEMATICAL CONSIDERATIONS ARE 6586 04:44:29,480 --> 04:44:31,360 IF YOU KNOW YOU HAVE AN 6587 04:44:31,360 --> 04:44:36,080 ANTAGONIST AND REFERENCE 6588 04:44:36,080 --> 04:44:38,760 ANTAGONIST YOU KNOW WHAT 100% 6589 04:44:38,760 --> 04:44:41,400 AND ASUM HUNDRED IS ANTAGONIST 6590 04:44:41,400 --> 04:44:43,760 AT BASELINE, WHEN IT DOESN'T GO 6591 04:44:43,760 --> 04:44:44,800 BASELINE IT IS NOT TYPICALLY 6592 04:44:44,800 --> 04:44:47,960 BECAUSE OF THE THE MAP, IT IS 6593 04:44:47,960 --> 04:44:50,760 BECAUSE COMPOUNDS SOLUTION, 6594 04:44:50,760 --> 04:44:53,000 THERE'S SOME REAL BIOLOGY TO 6595 04:44:53,000 --> 04:44:56,920 THAT. SO THING TO BE CAREFUL 6596 04:44:56,920 --> 04:44:57,680 ABOUT ARBITRARILY SAYING, 50% OF 6597 04:44:57,680 --> 04:44:58,920 TOTAL. 6598 04:44:58,920 --> 04:45:03,600 >>ARE YOU SAYING YOU PREFER 6599 04:45:03,600 --> 04:45:03,880 (INAUDIBLE)? 6600 04:45:03,880 --> 04:45:06,440 >>I'M DOING A GPCR LOOKING 6601 04:45:06,440 --> 04:45:09,600 FUNCTIONAL ASSAYS LOOKING EC 6602 04:45:09,600 --> 04:45:12,200 50s, I WOULD ALWAYS HAVE EC 50 6603 04:45:12,200 --> 04:45:14,600 AND A EFFICACY, TWO PARAMETER, 6604 04:45:14,600 --> 04:45:16,480 EFFICACY AND EC 50. 6605 04:45:16,480 --> 04:45:18,560 >>THAT IS WHAT I WAS SAYING SO 6606 04:45:18,560 --> 04:45:21,360 USE EC 50 ALONG WITH E MAX FOR 6607 04:45:21,360 --> 04:45:24,120 FUNCTIONAL ASSAYS. BUT I HAVE 6608 04:45:24,120 --> 04:45:25,680 SEEN SCENARIOS PEOPLE ARE 6609 04:45:25,680 --> 04:45:28,800 CONFLATE WITH THE ABSOLUTE EC 6610 04:45:28,800 --> 04:45:33,640 50. SO USING THAT NUMBER AND E 6611 04:45:33,640 --> 04:45:34,960 MAX TO GO WITH IT AND JUST 6612 04:45:34,960 --> 04:45:37,360 MAKING SURE COMPARING APPLES TO 6613 04:45:37,360 --> 04:45:38,760 AMOUNTS BY COMPARING TO THE 6614 04:45:38,760 --> 04:45:42,600 THRESHOLD OF 50%. AOC SEEMS TO 6615 04:45:42,600 --> 04:45:46,800 WORK WELL. SO WHEN DOING 6616 04:45:46,800 --> 04:45:52,640 MODELING, TO PREDICTION MODEL 6617 04:45:52,640 --> 04:45:56,600 AND REFINE SAR BASED ON THE QSAR 6618 04:45:56,600 --> 04:45:59,760 WHAT METRIC DO WE USE? EC 50 6619 04:45:59,760 --> 04:46:03,480 ABSOLUTE EC 50, E MAX? IN THAT 6620 04:46:03,480 --> 04:46:05,720 CASE I FOUND AOC WORKS WELL SO 6621 04:46:05,720 --> 04:46:08,160 WHEN YOU USE AOC AS RESPONSE TO 6622 04:46:08,160 --> 04:46:11,480 THEN BUILD PREDICTION MODEL QS 6623 04:46:11,480 --> 04:46:12,840 ARKANSAS MODEL, WE GET MORE 6624 04:46:12,840 --> 04:46:14,280 MEANINGFUL MODEL THAN JUST USING 6625 04:46:14,280 --> 04:46:15,520 A MODEL AGAINST ONE OF THE OTHER 6626 04:46:15,520 --> 04:46:19,560 PARAMETERS. 6627 04:46:19,560 --> 04:46:21,040 >>ALL RIGHT. THANK YOU VERY 6628 04:46:21,040 --> 04:46:22,960 MUCH, DEVAN FOR A WONDERFUL 6629 04:46:22,960 --> 04:46:23,520 TALK. 6630 04:46:23,520 --> 04:46:29,360 [APPLAUSE] 6631 04:46:29,360 --> 04:46:30,960 >>ALL RIGHT, EVERYONE, WE WILL 6632 04:46:30,960 --> 04:46:33,000 TAKE A BREAK NOW AND RECONVENE 6633 04:46:33,000 --> 04:46:34,320 AT 3:30. MAKE SURE YOU COME 6634 04:46:34,320 --> 04:46:36,520 BACK. THERE'S TWO AMAZING FINAL 6635 04:46:36,520 --> 04:46:38,000 TALKS TODAY. ONE IS ON DRUG 6636 04:46:38,000 --> 04:46:39,800 SCREENING WITH ENGINEERED TREATY 6637 04:46:39,800 --> 04:46:41,440 TISSUE MODELS. AND THE OTHER ONE 6638 04:46:41,440 --> 04:46:43,880 IS ON IN VITRO ASSESSMENT OF 6639 04:46:43,880 --> 04:46:46,040 PROPERTIES OF LINKED COMPOUNDS. 6640 04:46:46,040 --> 04:46:50,080 BE HERE AT 3:30 SO WE FINALIZE 6641 04:46:50,080 --> 04:46:50,800 WITH TWO AMAZING TALKS. ALL 6642 04:46:50,800 --> 04:46:52,800 RIGHT. HAVE A NICE BREAK. 6643 04:46:52,800 --> 04:46:55,560 OUR NEXT SPEAKER IS DR. MARC 6644 04:46:55,560 --> 04:46:57,960 FERRER, DR. FERRER IS CURRENTLY 6645 04:46:57,960 --> 04:47:01,600 THE DIRECTOR OF THE 3D TISSUE 6646 04:47:01,600 --> 04:47:03,800 BIOLABORATORY AT NCATS, A 6647 04:47:03,800 --> 04:47:05,280 MULTI-DISCIPLINARY GROUP 6648 04:47:05,280 --> 04:47:06,760 ESTABLISHING 2018 WITH GOAL OF 6649 04:47:06,760 --> 04:47:09,200 CREATING VALIDATING AND USING 6650 04:47:09,200 --> 04:47:12,120 TREATY BIOENGINEERED TISSUE FOR 6651 04:47:12,120 --> 04:47:13,240 DISEASE MODELING AND PREDICTIVE 6652 04:47:13,240 --> 04:47:15,800 DRUG DISCOVERY AND DEVELOPMENT. 6653 04:47:15,800 --> 04:47:17,960 HIS TALK IS ENTITLED DRUG 6654 04:47:17,960 --> 04:47:20,680 SCREENING WITH ENGINEERED 3D 6655 04:47:20,680 --> 04:47:23,040 TISSUE MODELS. THANK YOU FOR 6656 04:47:23,040 --> 04:47:33,560 BEING HERE. FLOOR IS YOURS. 6657 04:47:35,000 --> 04:47:36,200 THANK YOU FOR THE INTEREST AND 6658 04:47:36,200 --> 04:47:39,480 THANK YOU FOR STAYING. ALMOST TO 6659 04:47:39,480 --> 04:47:44,120 THE END. THE LAST TWO DAYS WE 6660 04:47:44,120 --> 04:47:45,640 LEARNED THAT DRUG DISCOVERY 6661 04:47:45,640 --> 04:47:47,040 PROBLEM YOU NEED TO HAVE A 6662 04:47:47,040 --> 04:47:51,000 BATTERY OF ASSAYS NOT ONLY A 6663 04:47:51,000 --> 04:47:52,880 PRIMARY ASSAY BUT SECONDARY 6664 04:47:52,880 --> 04:47:58,520 ASSAY FOR COUNTER SCREEN, FOR 6665 04:47:58,520 --> 04:48:00,960 READ OUTS.S. THESE NEED TO BE 6666 04:48:00,960 --> 04:48:03,680 ROBUST. THE FACTORS ARE MORE 6667 04:48:03,680 --> 04:48:09,080 THAN .5 AND MSR LESS THAN 3. SO 6668 04:48:09,080 --> 04:48:13,560 I WILL TRY TO CONVINCE YOU MORE 6669 04:48:13,560 --> 04:48:14,280 ASSAYS U YOU NEED ASSAYS THAT 6670 04:48:14,280 --> 04:48:15,760 ARE PHYSIOLOGICALLY RELEVANT. 6671 04:48:15,760 --> 04:48:17,040 THE REASON FOR THAT, WITH ALL 6672 04:48:17,040 --> 04:48:19,360 THESE ROBUST ASSAYS AND 6673 04:48:19,360 --> 04:48:21,080 BEAUTIFUL DOSE RESPONSES YOU 6674 04:48:21,080 --> 04:48:25,960 HAVE SEEN THESE TWO DAYS THE 6675 04:48:25,960 --> 04:48:28,520 RATE DRUGS GET APPROVED IS STILL 6676 04:48:28,520 --> 04:48:32,920 10% SO WHETHER TARGET 6677 04:48:32,920 --> 04:48:33,720 VALIDATION, WRONG TARGETS NOT 6678 04:48:33,720 --> 04:48:35,280 CLINICALLY VALIDATED OR THE 6679 04:48:35,280 --> 04:48:37,920 ASSAYS WE USE ARE TOO SIMPLISTIC 6680 04:48:37,920 --> 04:48:40,440 AND DON'T CAPTURE THE 6681 04:48:40,440 --> 04:48:42,200 ENVIRONMENT OF THE TARGET IN THE 6682 04:48:42,200 --> 04:48:46,160 ACTUAL BODY SO BEAUTIFUL TALKS 6683 04:48:46,160 --> 04:48:48,760 ALONG THE DAY WE USE ENZYMATIC 6684 04:48:48,760 --> 04:48:50,440 ASSAYS AND MANY TIMES YOU SCREEN 6685 04:48:50,440 --> 04:48:54,040 WITH CATALYTIC SIDE BUZZ FULL 6686 04:48:54,040 --> 04:49:03,720 LENGTH IS NOT MOST GPCR PROBABLY 6687 04:49:03,720 --> 04:49:04,760 SCREENING ARE CHOICE HEALTH SO 6688 04:49:04,760 --> 04:49:07,000 THAT IS NOT THE CELLS IN YOUR 6689 04:49:07,000 --> 04:49:10,080 BODY SO WE THINK PART OF THE 6690 04:49:10,080 --> 04:49:12,760 PROBLEM IS THAT BY CREATING 6691 04:49:12,760 --> 04:49:15,600 THESE ASAYS THAT ARE SO ROBUST, 6692 04:49:15,600 --> 04:49:18,600 WE MAKE COMPROMISES THE 6693 04:49:18,600 --> 04:49:20,600 COMPROMISE IS THEY LOSE THAT 6694 04:49:20,600 --> 04:49:22,120 PHYSIOLOGICAL RELEVANCE THAT WE 6695 04:49:22,120 --> 04:49:24,160 NEED IN THESE ASSAYS AND TOUCH 6696 04:49:24,160 --> 04:49:25,840 ON ASPECT OF BIOLOGYCAL 6697 04:49:25,840 --> 04:49:28,080 RELEVANCE BUT THE TRUTH IS THAT 6698 04:49:28,080 --> 04:49:31,680 BY GROWING CELLS ON GLASS NOT 6699 04:49:31,680 --> 04:49:33,360 PHYSIOLOGICALLY RELEVANT THE 6700 04:49:33,360 --> 04:49:34,280 CELLS IN BODY GROW TALKING TO 6701 04:49:34,280 --> 04:49:38,040 OTHER CELL TYPES, THEY GROW WITH 6702 04:49:38,040 --> 04:49:38,840 TYPES DIFFERENT (INAUDIBLE) 6703 04:49:38,840 --> 04:49:43,520 THERE'S MECHANICAL FORCES, THE 6704 04:49:43,520 --> 04:49:46,040 VASES QUEUE LATURE. SO SIGNALING 6705 04:49:46,040 --> 04:49:48,640 GENE EXPRESSION CELLS NEW TARGET 6706 04:49:48,640 --> 04:49:50,840 IN BODY IS PROBABLY NOT WHAT YOU 6707 04:49:50,840 --> 04:49:53,800 FIND IN OVEREXPRESSING THE 6708 04:49:53,800 --> 04:49:57,560 FAVORITE GPCR IN CELLS WHICH YOU 6709 04:49:57,560 --> 04:49:59,360 OPTIMIZE TO HAVE A WHOPPING 6710 04:49:59,360 --> 04:50:01,600 SIGNAL AND GOOD Z FACTOR AND 6711 04:50:01,600 --> 04:50:02,160 REPRODUCIBILITY AS YOU HEARD 6712 04:50:02,160 --> 04:50:07,320 BEFORE. SO WE ARE NOT THE ONLY 6713 04:50:07,320 --> 04:50:11,720 ONES, YOU PROBABLY HEARD ABOUT 6714 04:50:11,720 --> 04:50:14,520 3D MODELS TISSUE IN A CHIP, 6715 04:50:14,520 --> 04:50:18,160 ORGANOIDS STEROID, REALLY A 6716 04:50:18,160 --> 04:50:22,120 BROAD RANGE OF DIFFERENT 3D 6717 04:50:22,120 --> 04:50:25,680 MODELS AND THE IDEA IS THAT BY 6718 04:50:25,680 --> 04:50:29,560 CREATING THESE ASSAYS AND 6719 04:50:29,560 --> 04:50:31,360 PUTTING CELLS MORE TISSUE LIKE 6720 04:50:31,360 --> 04:50:34,240 DRUG RESPONSE WILL BE MORE LIKE 6721 04:50:34,240 --> 04:50:38,320 IN YOUR BODY. AS ALWAYS THIS 6722 04:50:38,320 --> 04:50:40,640 COMPROMISES WE CAN MAKE IT AS 6723 04:50:40,640 --> 04:50:42,880 COMPLEX AS YOU CAN OR SIMPLE AS 6724 04:50:42,880 --> 04:50:45,240 YOU CAN. SIMPLICITY BRINGS 6725 04:50:45,240 --> 04:50:46,800 THROUGH PUT WHICH IS IMPORTANT 6726 04:50:46,800 --> 04:50:50,760 FOR THE WORK THAT WE ARE DOING. 6727 04:50:50,760 --> 04:50:52,200 AS YOU INCREASE FIZZ CROW 6728 04:50:52,200 --> 04:50:55,800 LOGICAL COMPLEXITY, YOUR THROUGH 6729 04:50:55,800 --> 04:50:59,200 PUT GOES DOWN BECAUSE THESE ARE 6730 04:50:59,200 --> 04:51:00,480 TECHNICALLY COMPLEX TISSUE 6731 04:51:00,480 --> 04:51:03,280 MODELS WHERE YOU HAVE FLUIDICS, 6732 04:51:03,280 --> 04:51:06,600 YOU HAVE MUCH OF THE MODELS ARE 6733 04:51:06,600 --> 04:51:07,760 PHYSIOLOGICALLY MORE RELEVANT, 6734 04:51:07,760 --> 04:51:10,200 THEY ARE DEVELOPED WITH PRIMARY 6735 04:51:10,200 --> 04:51:12,720 CELLS, OR IPS DERIVED CELLS CELE 6736 04:51:12,720 --> 04:51:14,280 TRY TO AVOID THE CELL LINES THAT 6737 04:51:14,280 --> 04:51:18,560 WE USE IN NORMAL 2D BECAUSE 6738 04:51:18,560 --> 04:51:20,320 THESE CELLS HAVE PASSAGE IN 6739 04:51:20,320 --> 04:51:22,160 PLASTIC AND NOT WHAT YOU FIND IN 6740 04:51:22,160 --> 04:51:26,200 YOUR BODY. SO I WON'T GO INTO 6741 04:51:26,200 --> 04:51:27,560 DEAIL TO THE DIFFERENCES 6742 04:51:27,560 --> 04:51:29,600 SIMILARITIES PROS AND CONS, WE 6743 04:51:29,600 --> 04:51:33,560 HAVE A REVIEW MANY REVIEWERS BUT 6744 04:51:33,560 --> 04:51:34,880 WE HAD TABLE THAT EXPLAINS THE 6745 04:51:34,880 --> 04:51:39,000 DIFFERENCES IN SIMILARITIES WITH 6746 04:51:39,000 --> 04:51:42,520 DIFFERENT MODELS. WHAT IS 6747 04:51:42,520 --> 04:51:44,760 REALLY CRITICAL TO ALL OF THEM 6748 04:51:44,760 --> 04:51:46,680 IS THE QUESTION YOU ALWAYS GET, 6749 04:51:46,680 --> 04:51:50,440 HOW PREDICTABLE -- CLINICALLY 6750 04:51:50,440 --> 04:51:51,440 PREDICTIVE ARE THESE MODELS 6751 04:51:51,440 --> 04:51:54,600 COMPARED TO THE 2D MODEL. THIS 6752 04:51:54,600 --> 04:51:55,640 IS SOMETHING YOU HAVE TO KEEP IN 6753 04:51:55,640 --> 04:51:59,680 MIND AS YOU START DEVELOPING ANY 6754 04:51:59,680 --> 04:52:02,880 OF THESE 3D MODELS ORGANOIDS THE 6755 04:52:02,880 --> 04:52:04,840 OTHER ASPECT IS HOW MUCH 6756 04:52:04,840 --> 04:52:05,920 PHYSIOLOGICAL COMPLEXITY DO YOU 6757 04:52:05,920 --> 04:52:08,640 NEED TO INCLUDE IN THESE 6758 04:52:08,640 --> 04:52:11,200 MODELINGS TO HAVE THE 6759 04:52:11,200 --> 04:52:12,080 PREDICTABILITYIBILITY YOU ARE 6760 04:52:12,080 --> 04:52:14,640 LOOKING FOR FOR A SPECIFIC 6761 04:52:14,640 --> 04:52:15,680 DISEASE, SPECIFIC TARGET YOU ARE 6762 04:52:15,680 --> 04:52:17,080 INTERESTED IN. WHAT CELL TYPES 6763 04:52:17,080 --> 04:52:18,600 DO YOU NEED TO INCLUDE IN YOUR 6764 04:52:18,600 --> 04:52:23,520 MODEL? HOW YOU HAVE ALL THESE 6765 04:52:23,520 --> 04:52:24,920 DIFFERENT CELL TYPES AND WHAT 6766 04:52:24,920 --> 04:52:27,200 ARE YOU GOING TO USE, EVERY CELL 6767 04:52:27,200 --> 04:52:30,040 YOU BUY FROM A DIFFERENT VENDOR, 6768 04:52:30,040 --> 04:52:32,520 COMES WITH DIFFERENT MAGIC 6769 04:52:32,520 --> 04:52:34,160 MEDIA, SOMETIMES YOU DON'T KNOW 6770 04:52:34,160 --> 04:52:37,520 WHAT IS IN THE MEDIA. HOW YOU 6771 04:52:37,520 --> 04:52:40,000 BRING THESE TOGETHER MAKE SYSTEM 6772 04:52:40,000 --> 04:52:41,600 3DISH TISSUE SYSTEM THAT IS 6773 04:52:41,600 --> 04:52:43,880 FUNCTIONING BY WHATEVER CRITERIA 6774 04:52:43,880 --> 04:52:47,760 YOU DECIDE TO HAVE. AND AS YOU 6775 04:52:47,760 --> 04:52:50,280 KNOW, MANY DIFFERENT PLATFORMS, 6776 04:52:50,280 --> 04:52:53,120 DIFFERENT VENDORS HOW DO YOU 6777 04:52:53,120 --> 04:52:55,880 CHOOSE? AT THE END OF THE DAY 6778 04:52:55,880 --> 04:52:57,880 DECIDING BASED ON APPLICATION 6779 04:52:57,880 --> 04:52:58,800 AND THE QUESTIONS YOU ARE TRYING 6780 04:52:58,800 --> 04:53:03,800 TO ANSWER. YOU HEARD ABOUT 6781 04:53:03,800 --> 04:53:06,000 VALIDATION. BIOLOGICAL 6782 04:53:06,000 --> 04:53:07,320 VALIDATION, IN OUR CASE, YOU 6783 04:53:07,320 --> 04:53:11,240 WILL SEE IN EXAMPLES, I WILL 6784 04:53:11,240 --> 04:53:13,920 SHOW WE ARE TALKING ABOUT CELL 6785 04:53:13,920 --> 04:53:15,720 MARKERS WHAT CELLS DO I HAVE IN 6786 04:53:15,720 --> 04:53:19,600 MY TISSUE, DO THEY EXPRESS SAME 6787 04:53:19,600 --> 04:53:22,520 MARKERS THAT I WOULD EXPECT IN 6788 04:53:22,520 --> 04:53:25,440 THIS SAME CELLS IN A HUMAN 6789 04:53:25,440 --> 04:53:28,320 TISSUE. CELL COMPOSITION 6790 04:53:28,320 --> 04:53:30,760 MORPHOLOGY, DOES THE TISSUE HAVE 6791 04:53:30,760 --> 04:53:35,520 MORPHOLOGY YOU WOULD EXPECT IN A 6792 04:53:35,520 --> 04:53:37,560 HUMAN. WE DO A LOT OF 6793 04:53:37,560 --> 04:53:39,720 PROTEOMICS. HOW YOU VALIDATE A 6794 04:53:39,720 --> 04:53:41,280 LOT OF IMMUNOHISTOCHEMISTRY, 6795 04:53:41,280 --> 04:53:43,200 IMAGING YOU WILL SEE. WHETHER IN 6796 04:53:43,200 --> 04:53:44,320 SINGLE CELL SEQUENCING BECAUSE 6797 04:53:44,320 --> 04:53:46,960 AT THE END OF THE DAY YOU WANT 6798 04:53:46,960 --> 04:53:50,040 TO BENCHMARK THESE TISSUES YOU 6799 04:53:50,040 --> 04:53:52,720 ARE MAKING TO THE CLINICAL 6800 04:53:52,720 --> 04:53:54,440 SAMPLES. WE WANT TO BE CLOSE TO 6801 04:53:54,440 --> 04:53:59,160 CLINIC, SHOW US THAT YOU CAN DO 6802 04:53:59,160 --> 04:53:59,720 IT HOY CLOSE BY MORPHOLOGY 6803 04:53:59,720 --> 04:54:01,560 FUNCTION OMICS. HOW CLOSE WILL 6804 04:54:01,560 --> 04:54:04,840 YOU TO THAT HUMAN SAMPLE. ONCE 6805 04:54:04,840 --> 04:54:06,520 YOU GET AND CONSEQUENCE YOURSELF 6806 04:54:06,520 --> 04:54:07,560 YOU HAVE A TISSUE WITH THE RIGHT 6807 04:54:07,560 --> 04:54:08,760 MARKERS, THE RIGHT FUNCTION, 6808 04:54:08,760 --> 04:54:11,840 RIGHT MORPHOLOGY, YOU WANT TO 6809 04:54:11,840 --> 04:54:13,320 USE THEM TO BE ASSAY TO TEST 6810 04:54:13,320 --> 04:54:17,640 COMPOUNDS. SO USER TO WORRY 6811 04:54:17,640 --> 04:54:20,200 ABOUT EVERYTHING SAID ABOUT Z 6812 04:54:20,200 --> 04:54:21,640 FACTOR DOS THEY HAVE ROBUSTNESS. 6813 04:54:21,640 --> 04:54:23,240 REPRESENTATIVE FROM DAY TO DAY 6814 04:54:23,240 --> 04:54:26,120 WELL TO WELL PLATE TO PLATE. 6815 04:54:26,120 --> 04:54:28,680 MANY TIMES YOU USE EXPERIMENT 6816 04:54:28,680 --> 04:54:30,360 ITSELF SO A LOT OF PRIMARY CELLS 6817 04:54:30,360 --> 04:54:33,160 NO LONGER AVAILABLE I'M CHANGING 6818 04:54:33,160 --> 04:54:35,720 LOGS DO I GET THE SAME? THE 6819 04:54:35,720 --> 04:54:37,320 ANSWER IS NO. YOU DON'T KNOW 6820 04:54:37,320 --> 04:54:39,720 WHY. BUT YOU HAVE TO ALMOST 6821 04:54:39,720 --> 04:54:42,920 RE-EVALUATE AND REDEVELOP ASSAY 6822 04:54:42,920 --> 04:54:43,560 FROM THE CRUTCH BECAUSE CELLS 6823 04:54:43,560 --> 04:54:46,880 WERE DIFFERENT. AGAIN WE WANT TO 6824 04:54:46,880 --> 04:54:50,000 USE THESE ASSAYS FOR SCREENING, 6825 04:54:50,000 --> 04:54:51,640 WE WANT TO DEVELOP FUNCTIONAL 6826 04:54:51,640 --> 04:54:53,800 END POINTS. YOU CANNOT DO A 6827 04:54:53,800 --> 04:54:55,760 SCREEN WITH SECTIONING AND 6828 04:54:55,760 --> 04:54:57,680 IMMUNOHISTOCHEMISTRY. YOU CAN DO 6829 04:54:57,680 --> 04:55:00,840 -- YOU NEED ASSAYS THAT ARE 6830 04:55:00,840 --> 04:55:02,880 AMENABLE TO THE ALTERATIONS AND 6831 04:55:02,880 --> 04:55:05,880 READERS THAT YOU USE TO REGULAR 6832 04:55:05,880 --> 04:55:09,440 2D ASSAYS SO WHAT TYPE OF ASSAY 6833 04:55:09,440 --> 04:55:11,680 READ OUTS WE USE, WE USE A LOT 6834 04:55:11,680 --> 04:55:14,520 OF HIGH CONTENT SCREEN, USE A 6835 04:55:14,520 --> 04:55:17,760 LOT OF ASSAYS THAT PERHAPS YOU 6836 04:55:17,760 --> 04:55:21,080 CAN SEW IN A 1536 WELL PLATE BUT 6837 04:55:21,080 --> 04:55:22,560 FOR THIS TYPE OF THROUGH PUT, 6838 04:55:22,560 --> 04:55:26,200 IT'S OKAY TO WANT ELISA, 6839 04:55:26,200 --> 04:55:28,400 ACTUALLY POWERFUL SO WE DO 6840 04:55:28,400 --> 04:55:31,080 MULTIPLEX ELISA, LUMINEX TYPE OF 6841 04:55:31,080 --> 04:55:32,560 ASSAYS. THE HOPE IS THAT WE WILL 6842 04:55:32,560 --> 04:55:34,600 BE ABLE TO APPLY THE SAME 6843 04:55:34,600 --> 04:55:36,480 ROBUSTNESS CRITERIA THAT TC 6844 04:55:36,480 --> 04:55:39,400 DESCRIBED IN HIS TALK CAN WE 6845 04:55:39,400 --> 04:55:41,640 MAKE ASSAYS IN TISSUE MODELS AND 6846 04:55:41,640 --> 04:55:43,320 ENGINEER TISSUE MODELS WITH Z 6847 04:55:43,320 --> 04:55:47,320 FACTOR .5. I WILL SHOW IN SOME 6848 04:55:47,320 --> 04:55:51,880 ASSAYS WE CAN GIVE THEM. AND 6849 04:55:51,880 --> 04:55:55,320 FINALLY, AS DEVAN WHEN IN THE 6850 04:55:55,320 --> 04:55:56,560 LAST TALK TAKING CONTROL 6851 04:55:56,560 --> 04:55:58,000 COMPOUNDS AND DOING IC 50 AND 6852 04:55:58,000 --> 04:55:59,800 LOOKING AT POTENCY AND EFFICACY 6853 04:55:59,800 --> 04:56:00,960 OF COMPOUNDS IN THESE TISSUE 6854 04:56:00,960 --> 04:56:04,480 MODELS AND COMPARING IT TO 2D, 6855 04:56:04,480 --> 04:56:06,240 SO WE ALWAYS WE DON'T WANT MORE 6856 04:56:06,240 --> 04:56:07,200 WORK ON OURSELVES IF WE DON'T 6857 04:56:07,200 --> 04:56:13,400 HAVE TO. WE TRY TO BENCHMARK 6858 04:56:13,400 --> 04:56:16,080 TISSUES AGAINST CELLS IN 2D. 6859 04:56:16,080 --> 04:56:20,720 ADVANTAGE TO TAKE IPS DERIVED 6860 04:56:20,720 --> 04:56:23,200 NEWSPAPER AND PUT IN CONTEXT OF 6861 04:56:23,200 --> 04:56:24,200 TISSUE VERSUS SCREEN NEURON IN 6862 04:56:24,200 --> 04:56:32,320 2D. YOU HAVE TO GO BACK AND 6863 04:56:32,320 --> 04:56:33,880 BENCHMARK TO SEE ADVANTAGE OR 6864 04:56:33,880 --> 04:56:36,240 NOT. NOT ONLY IN THE BIOLOGY, 6865 04:56:36,240 --> 04:56:38,720 BUT ALSO IN THE PHARMACOLOGY OF 6866 04:56:38,720 --> 04:56:43,880 THE COMPOUNDS. WE DECIDED TO 6867 04:56:43,880 --> 04:56:46,200 INVEST IN 3D TISSUE 6868 04:56:46,200 --> 04:56:50,560 BIOFABRICATION. IT IS REALLY AN 6869 04:56:50,560 --> 04:56:52,240 EMERGING TECHNOLOGY IS THAT IS 6870 04:56:52,240 --> 04:56:56,240 POWERFUL AND ALLOWS YOU TO 6871 04:56:56,240 --> 04:56:59,320 ASSEMBLE COMPLEX ISSUES WITH A 6872 04:56:59,320 --> 04:57:02,280 SPECIAL CONTROL WHATEVER CELL 6873 04:57:02,280 --> 04:57:03,080 COMPOSITION YOU WANT TO INCLUDE 6874 04:57:03,080 --> 04:57:04,800 IN YOUR TISSUE. SO IT IS 6875 04:57:04,800 --> 04:57:09,680 ADDITIVE MANUFACTURING. FOR US 6876 04:57:09,680 --> 04:57:13,000 IT WAS INTERESTING BECAUSE WE 6877 04:57:13,000 --> 04:57:14,760 COULD HAVE SPATIAL CONTROL 6878 04:57:14,760 --> 04:57:19,240 MULTI-CELL TYPE STRUCTURES IF 6879 04:57:19,240 --> 04:57:22,200 YOU WANT A DISEASE TISSUE WE CAN 6880 04:57:22,200 --> 04:57:24,960 CHOOSE WHAT CELLS WE INCLUDE 6881 04:57:24,960 --> 04:57:26,680 NORMAL VERSUS DISEASE AND 6882 04:57:26,680 --> 04:57:27,720 UNDERSTAND A LITTLE BIT WHAT 6883 04:57:27,720 --> 04:57:30,440 TYPE OF CELLS IN THAT PARTICULAR 6884 04:57:30,440 --> 04:57:32,560 TISSUE WAS SEPARATING THE 6885 04:57:32,560 --> 04:57:36,320 DISEASE MODELS OR PHENOTYPE. 6886 04:57:36,320 --> 04:57:38,760 BIOPRINTING ALLOWS PHYSIOLOGICAL 6887 04:57:38,760 --> 04:57:40,240 FEATURES LIKE VASCULATURE, 6888 04:57:40,240 --> 04:57:43,760 INCLUDE IMMUNE CELLS, SO VERY 6889 04:57:43,760 --> 04:57:44,640 VERSATILE TECHNOLOGY TERMS OF 6890 04:57:44,640 --> 04:57:47,680 BEING ABLE TO INCLUDE WHATEVER 6891 04:57:47,680 --> 04:57:48,480 PHYSIOLOGY YOU WANT TO INCLUDE 6892 04:57:48,480 --> 04:57:49,280 IN YOUR PRESENTATION. MOST OF 6893 04:57:49,280 --> 04:57:52,000 THE TIME WE USE PRIMARY CELLS OR 6894 04:57:52,000 --> 04:57:54,360 IPS DERIVED CELLS, THE TIME TO 6895 04:57:54,360 --> 04:57:57,000 DEVELOPMENT AND HAVING MATURE 6896 04:57:57,000 --> 04:57:58,840 TISSUE IS SHORTER THAN PERHAPS 6897 04:57:58,840 --> 04:58:01,920 MAYBE ORGANOIDS FOR THOSE OF YOU 6898 04:58:01,920 --> 04:58:05,000 DEVELOP ORGANOIDS MIGHT TAKE 6899 04:58:05,000 --> 04:58:05,800 WEEKS AND NON-HAVE MATURE 6900 04:58:05,800 --> 04:58:08,000 ORGANOID, THAT IS FUNCTIONAL. 6901 04:58:08,000 --> 04:58:09,960 HERE WE CAN GO THREE TO FOUR 6902 04:58:09,960 --> 04:58:13,320 WEEKS, STILL LONG TIME, COMPARED 6903 04:58:13,320 --> 04:58:15,200 TO 2D MODEL BUT IT IS RELATIVELY 6904 04:58:15,200 --> 04:58:17,160 SHORT WHEN YOU TALK 3D MODEL. 6905 04:58:17,160 --> 04:58:21,960 WE CAN DO THAT THIS TISSUE IN A 6906 04:58:21,960 --> 04:58:23,920 REPRODUCIBLE WAY UP TO 96 WELL 6907 04:58:23,920 --> 04:58:27,120 PLACE, WHICH IS FOR US HIGH 6908 04:58:27,120 --> 04:58:32,240 THROUGH PUT. SO BIOPRINTING 101, 6909 04:58:32,240 --> 04:58:34,200 IT IS USED, ADDITIVE 6910 04:58:34,200 --> 04:58:36,560 MANUFACTURING TECHNOLOGY, 6911 04:58:36,560 --> 04:58:37,360 PROBABLY HAVE BEEN USED FOR 6912 04:58:37,360 --> 04:58:40,560 PRODUCTION OF ORGANS OR 6913 04:58:40,560 --> 04:58:41,960 REGENERATIVE MEDICINE. WE USE IT 6914 04:58:41,960 --> 04:58:44,240 TO CREATE THE COMPLEX 3D TISSUE 6915 04:58:44,240 --> 04:58:48,560 MODELS. THIS NOW SEVERAL BRANDS 6916 04:58:48,560 --> 04:58:50,640 OF BIOPRINTERS IN THE MARKERS SO 6917 04:58:50,640 --> 04:58:53,720 COMMERCIALLY AVAILABLE. THEY 6918 04:58:53,720 --> 04:58:55,880 RANGE FROM 50,000 TO HALF 6919 04:58:55,880 --> 04:58:59,720 MILLION DOLLARS DEPENDING ON 6920 04:58:59,720 --> 04:59:00,960 FEATURES YOU WANT IN THE 6921 04:59:00,960 --> 04:59:04,840 BIOPRINT BUT MOST HAVE LIKE 6922 04:59:04,840 --> 04:59:09,320 DROPLETS OF CELLS. HAVE 6923 04:59:09,320 --> 04:59:10,920 EXTRUSION YOU MIX YOUR CELLS 6924 04:59:10,920 --> 04:59:13,360 WITH HYDRA GEL AND PRESSURIZE 6925 04:59:13,360 --> 04:59:17,240 SQUEEZE YOUR KETCHUP OR MISON 6926 04:59:17,240 --> 04:59:18,800 YOUR HOT DOG. AT THE END OF THE 6927 04:59:18,800 --> 04:59:21,480 DAY, THE TECHNOLOGY WE USE IS 6928 04:59:21,480 --> 04:59:24,440 REALLY HYDRO GEL BASED EXTRUSION 6929 04:59:24,440 --> 04:59:26,240 FOR MOST OF OUR TISSUES. WHAT 6930 04:59:26,240 --> 04:59:32,640 YOU DO HERE IS MIX YOUR CELLS OF 6931 04:59:32,640 --> 04:59:35,720 INTEREST WITH HYDRO GEL. WE USE 6932 04:59:35,720 --> 04:59:39,120 GELATIN BASE WE ADD COLLAGEN, A 6933 04:59:39,120 --> 04:59:40,400 LOT OF DIFFERENT GELS BEING 6934 04:59:40,400 --> 04:59:42,600 DEVELOPED AT THE MOMENT BUT THE 6935 04:59:42,600 --> 04:59:45,360 IDEA IS THAT THIS HYDRO GEL HAS 6936 04:59:45,360 --> 04:59:46,440 TO HAVE THE RIGHT IMMUNOLOGICAL 6937 04:59:46,440 --> 04:59:48,320 PROPERTIES SO YOU CAN SQUEEZE IT 6938 04:59:48,320 --> 04:59:52,280 THROUGH MIDDLE OF YOUR SYRINGE. 6939 04:59:52,280 --> 04:59:53,640 SO YOU DON'T WANT TO KILL CELLS 6940 04:59:53,640 --> 04:59:56,000 BY EXTRUDING THEM THROUGH THE 6941 04:59:56,000 --> 04:59:58,440 NEEDLE. IT HAS TO PROMOTE CELL 6942 04:59:58,440 --> 04:59:59,280 ADHESION, YOU DON'T WANT CELLS 6943 04:59:59,280 --> 05:00:02,280 TO BE FLOATING IN THE GEL. YOU 6944 05:00:02,280 --> 05:00:08,480 WANT CELLS ATTACHING TO THE GEL. 6945 05:00:08,480 --> 05:00:10,720 NON-TOXIC AND PRINTABLE 6946 05:00:10,720 --> 05:00:12,120 PROPERTIES, IN MANY CASES YOU 6947 05:00:12,120 --> 05:00:16,320 WANT YOUR HYDRO GEL TO BE 6948 05:00:16,320 --> 05:00:17,960 BIODEGRADABLE, YOU WANT IT TO 6949 05:00:17,960 --> 05:00:19,360 MAKE A STRUCTURE WITH THE 6950 05:00:19,360 --> 05:00:21,160 PRINTER BUT YOU WANT THE CELLS 6951 05:00:21,160 --> 05:00:23,240 TO -- IF YOU MAKE A TISSUE AND 6952 05:00:23,240 --> 05:00:25,760 WANT THE CELLS IN YOUR TISSUE TO 6953 05:00:25,760 --> 05:00:27,640 BE SECRETING NATIVE EXTRA 6954 05:00:27,640 --> 05:00:29,960 CELLULAR MATRIX. THAT'S THE 6955 05:00:29,960 --> 05:00:32,480 ULTIMATE GOAL SO YOU HAVE A 6956 05:00:32,480 --> 05:00:37,280 BALANCE BETWEEN YOUR GELATIN OR 6957 05:00:37,280 --> 05:00:39,840 FIBRIN TO CREATE AT THE SAME 6958 05:00:39,840 --> 05:00:41,640 TIME THE TISSUE MAKES ONLY ECM 6959 05:00:41,640 --> 05:00:43,720 SO LOOKING AGENT THE RIGHT 6960 05:00:43,720 --> 05:00:47,200 BALANCE ESSENTIALLY IF THE GELS 6961 05:00:47,200 --> 05:00:50,320 DEGRADE TOO FAST YOU GET PANCAKE 6962 05:00:50,320 --> 05:00:51,440 BECAUSE EVERYTHING FLATTENS. SO 6963 05:00:51,440 --> 05:00:55,080 IT IS AN ART TO THE ASSAY IN THE 6964 05:00:55,080 --> 05:00:58,960 TISSUE, TO GET GOOD SPATIAL 6965 05:00:58,960 --> 05:01:00,680 CONTROL REPRODUCIBILITY AND DO 6966 05:01:00,680 --> 05:01:03,760 MEASURES. THIS IS JUST SOME 6967 05:01:03,760 --> 05:01:08,320 MOVIES, EXTRUSION BASE, THIS IS 6968 05:01:08,320 --> 05:01:11,040 -- THIS WAS SOME SORT OF INITIAL 6969 05:01:11,040 --> 05:01:12,920 EXPERIMENTS WE KID TO 6970 05:01:12,920 --> 05:01:14,280 DEMONSTRATE YOU CAN -- WE DID TO 6971 05:01:14,280 --> 05:01:15,880 DEMONSTRATE YOU CAN CREATE 6972 05:01:15,880 --> 05:01:19,200 PATTERNS WITH A COMPUTER DESIGN 6973 05:01:19,200 --> 05:01:22,680 SOFTWARE TO 2D SO WE MADE THIS 6974 05:01:22,680 --> 05:01:26,800 PIZZA WITH RED AND GREEN 6975 05:01:26,800 --> 05:01:28,920 FIBROBLASTS. YOU CAN MAKE 6976 05:01:28,920 --> 05:01:30,200 SPATIAL ARRANGEMENTS. CELLS 6977 05:01:30,200 --> 05:01:32,520 THAT ARE LIVE AND DO IT ALSO IN 6978 05:01:32,520 --> 05:01:35,000 3D SO THIS IS IS THE SHAPES WE 6979 05:01:35,000 --> 05:01:37,720 DID AT THE BEGINNING WHEN WE 6980 05:01:37,720 --> 05:01:39,560 STARTED WORKING USING THE 6981 05:01:39,560 --> 05:01:41,000 BIOPRINTER SO I WILL TELL YOU 6982 05:01:41,000 --> 05:01:42,920 ABOUT TWO MODELS THAT WE HAVE 6983 05:01:42,920 --> 05:01:44,400 BEEN WORK, ONE IS SKIN, THE 6984 05:01:44,400 --> 05:01:46,960 OTHER ONE IS A NEUROVASCULAR 6985 05:01:46,960 --> 05:01:48,120 UNIT THAT WE HAVE BEEN WORKING. 6986 05:01:48,120 --> 05:01:50,440 WE HAVE BEEN USING TO GENERATE A 6987 05:01:50,440 --> 05:01:56,080 MODEL OF GLIOM GLIOMA. SKIN ON 6988 05:01:56,080 --> 05:01:59,560 SIMPLISTIC WAY YOUR DERMIS WHICH 6989 05:01:59,560 --> 05:02:05,960 IS LOWER LAYER WHICH IS MOSTLY 6990 05:02:05,960 --> 05:02:08,120 FIBROBLASTS AND ECM BUT MAKE IT 6991 05:02:08,120 --> 05:02:10,440 AS COMPLEX AS YOU WANT WITH 6992 05:02:10,440 --> 05:02:15,080 VASCULATURE, WITH HAIR, WITH 6993 05:02:15,080 --> 05:02:16,600 NERVES AND SWEAT GLANDS AND 6994 05:02:16,600 --> 05:02:20,800 EPIDERMIS ON TOP IS ESSENTIALLY 6995 05:02:20,800 --> 05:02:22,040 KERATINOCYTES WHICH HAVE BEEN 6996 05:02:22,040 --> 05:02:23,200 STRATIFIED IN DIFFERENTIATED 6997 05:02:23,200 --> 05:02:28,600 WHEN EXPOSED TO THE INTERFACE SO 6998 05:02:28,600 --> 05:02:29,920 MANY YEARS PEOPLE HAVE BEEN 6999 05:02:29,920 --> 05:02:31,720 DEVELOPING PROTOCOLS TO MAKE 7000 05:02:31,720 --> 05:02:36,160 SKIN EQUIVALENTS BY USING THUD 7001 05:02:36,160 --> 05:02:41,920 TRANSWELL PLACE WITH INSERTS AND 7002 05:02:41,920 --> 05:02:46,360 YOU CAN MAKE -- SO YOU PUT YOUR 7003 05:02:46,360 --> 05:02:49,880 SKIN TISSUE OVER DERMIS WITH 7004 05:02:49,880 --> 05:02:51,880 FIBROBLASTS AND YOU ADD A LAYER 7005 05:02:51,880 --> 05:02:55,960 OF KERATINOCYTES AND YOU DO 7006 05:02:55,960 --> 05:02:57,440 SUBMERGE AFTER A FEW DAYS ONCE 7007 05:02:57,440 --> 05:02:58,960 FORMS THE TISSUE YOU BRING THE 7008 05:02:58,960 --> 05:03:01,640 WHOLE TISSUE TO LIQUID 7009 05:03:01,640 --> 05:03:02,440 INTERFACE, ESSENTIALLY YOU 7010 05:03:02,440 --> 05:03:03,920 REMOVE THE MEDIA FROM THE TOP 7011 05:03:03,920 --> 05:03:05,960 AND KERATINOCYTES DIFFERENTIATE 7012 05:03:05,960 --> 05:03:09,760 MAKING YOUR EPIDERMIS. SO WITH 7013 05:03:09,760 --> 05:03:13,320 THIS BASIC PROTOCOLS WE CREATED 7014 05:03:13,320 --> 05:03:16,960 A PLATFORM OF DIFFERENT SKIN 7015 05:03:16,960 --> 05:03:18,880 TISSUE. HERE IS WHERE 7016 05:03:18,880 --> 05:03:20,560 INVESTIGATING WHAT PHYSIOLOGICAL 7017 05:03:20,560 --> 05:03:24,400 COMPLEXITY DO I NEED IN MY MODEL 7018 05:03:24,400 --> 05:03:25,960 FOR WHATEVER DISEASE, WHATEVER 7019 05:03:25,960 --> 05:03:29,080 BIOLOGY I WANT TO STUDY SO YOU 7020 05:03:29,080 --> 05:03:34,520 CAN MAKE SKIN EPIDERMIS BY 7021 05:03:34,520 --> 05:03:35,880 KERATINOCYTES PUTTING IN RIGHT 7022 05:03:35,880 --> 05:03:38,720 MEDIA AND EARLY INTERFACE. YOU 7023 05:03:38,720 --> 05:03:44,880 CAN MAKE FULL SKIN, USING A 7024 05:03:44,880 --> 05:03:47,240 BIOPRINTER, BY MIXING 7025 05:03:47,240 --> 05:03:50,280 FIBROBLASTS WITH KERATINOCYTES, 7026 05:03:50,280 --> 05:03:51,920 WE HAVE'S SKYLARIZED DERMIS BY 7027 05:03:51,920 --> 05:03:58,400 ADDING ENDOTHELIAL CELLS 7028 05:03:58,400 --> 05:04:00,440 RECENTLY WE HAVE BEEN WORKING ON 7029 05:04:00,440 --> 05:04:04,520 INCLUDING IMMUNE CELLS IN 7030 05:04:04,520 --> 05:04:06,120 MACROPHAGES THROUGH ELEVATED 7031 05:04:06,120 --> 05:04:07,320 PHYSIOLOGICAL COMPLEXITY WE NEED 7032 05:04:07,320 --> 05:04:11,120 FOR MODELS WE ARE INTERESTED IN 7033 05:04:11,120 --> 05:04:13,000 AND AGAIN FOR ALL THESE MODELS 7034 05:04:13,000 --> 05:04:16,000 WE ARE DEVELOPING EXTENSIVE 7035 05:04:16,000 --> 05:04:19,840 CHARACTERIZATION USING HISTOLOGY 7036 05:04:19,840 --> 05:04:20,680 IMMUNOHISTOCHEMISTRY, USING A 7037 05:04:20,680 --> 05:04:23,680 LOT OF HIGH CONTENT IMAGING WITH 7038 05:04:23,680 --> 05:04:25,920 CONFOCAL MICROSCOPY. AT THE SAME 7039 05:04:25,920 --> 05:04:30,200 TIME AS MENTIONED BEFORE WE WANT 7040 05:04:30,200 --> 05:04:34,240 FUNCTIONAL -- CELL VIABILITY, 7041 05:04:34,240 --> 05:04:35,920 ELECTRICAL RESISTANCE 7042 05:04:35,920 --> 05:04:37,520 MEASUREMENTS. WE TAKE 7043 05:04:37,520 --> 05:04:39,360 SUPERNATANTS AND MEASURE 7044 05:04:39,360 --> 05:04:41,640 CYTOKINES, AND USE HIGH CONTENT 7045 05:04:41,640 --> 05:04:43,440 IMAGING AS I MENTIONED BEFORE. 7046 05:04:43,440 --> 05:04:45,760 WHAT IS NICE ABOUT THIS PLATFORM 7047 05:04:45,760 --> 05:04:49,040 IS THAT IT IS MODULAR, VERSATILE 7048 05:04:49,040 --> 05:04:52,240 SO WE DEVELOP NORMAL SKIN TISSUE 7049 05:04:52,240 --> 05:04:53,800 OF WHATEVER COMPLEXITY WE WANT 7050 05:04:53,800 --> 05:04:57,560 AND WE ADD DIFFERENT INSULTS TO 7051 05:04:57,560 --> 05:04:59,680 CREATE DISEASE MODEL. THE INSULT 7052 05:04:59,680 --> 05:05:02,640 CAN BE A CYTOKINE, A CANCER 7053 05:05:02,640 --> 05:05:06,400 CELL, IT CAN BE A VIRUS, OR IT 7054 05:05:06,400 --> 05:05:08,000 CAN BE IRRITANT IF YOU ARE 7055 05:05:08,000 --> 05:05:09,200 INTERESTED IN SPECIFIC -- BUT 7056 05:05:09,200 --> 05:05:10,720 THE BOTTOM LINE IS WHAT TAKES A 7057 05:05:10,720 --> 05:05:13,800 LOT OF TIME IS DEVELOPING THE 7058 05:05:13,800 --> 05:05:15,320 INITIAL TISSUE AND EVALUATING 7059 05:05:15,320 --> 05:05:19,560 THE INITIAL TISSUE. THEN IT IS 7060 05:05:19,560 --> 05:05:21,680 ADDING THAT DISEASE INSULT 7061 05:05:21,680 --> 05:05:24,880 NORMALLY DOESN'T TAKE AS LONG 7062 05:05:24,880 --> 05:05:26,000 THOUGH IT IS STILL AN ART IN 7063 05:05:26,000 --> 05:05:27,080 TERMS OF DEVELOPING THE RIGHT 7064 05:05:27,080 --> 05:05:28,960 CONDITIONS. SO TODAY I WILL 7065 05:05:28,960 --> 05:05:33,520 SHOW YOU AN EXAMPLE OF ECTOPIC 7066 05:05:33,520 --> 05:05:36,520 DERMATITIS MODEL, ECZEMA AND ONE 7067 05:05:36,520 --> 05:05:39,000 OF THE CANCER MODELS. SO WE HAVE 7068 05:05:39,000 --> 05:05:40,120 BEEN INTERESTED IN DEVELOPING 7069 05:05:40,120 --> 05:05:47,560 CANCER MODELS BEYOND TAKING 7070 05:05:47,560 --> 05:05:48,360 PROLIFERATION CELLS AND GROWING 7071 05:05:48,360 --> 05:05:51,360 THEM IN ASSAY. GROWING TUMORS IN 7072 05:05:51,360 --> 05:05:52,840 CONTEXT OF TISSUE WHERE THE 7073 05:05:52,840 --> 05:05:55,200 TUMOR GROWS SO WE ARE INTERESTED 7074 05:05:55,200 --> 05:05:56,240 IN SKIN MODEL SO WE ARE 7075 05:05:56,240 --> 05:05:59,560 INTERESTED IN LOOKING AT HOW DO 7076 05:05:59,560 --> 05:06:00,800 MELANOMA CANCER CELLS GROW IN 7077 05:06:00,800 --> 05:06:04,160 THE CONTEXT OF THE SKIN. OR 7078 05:06:04,160 --> 05:06:08,040 ANOTHER WE HEARD IS HOW DO SMALL 7079 05:06:08,040 --> 05:06:11,320 CELL CARCINOMA CANCER CELLS GROW 7080 05:06:11,320 --> 05:06:13,120 IN CONTEXT. THEN THE QUESTION 7081 05:06:13,120 --> 05:06:16,880 LIKE OKAY, SO IF YOU HAVE 7082 05:06:16,880 --> 05:06:23,160 COMPOUNDS THAT DO WE HAVE 7083 05:06:23,160 --> 05:06:25,480 COMPOUNDS THAT WORK MORE 7084 05:06:25,480 --> 05:06:27,760 PREDICTIVE IN TERMS OF THE 7085 05:06:27,760 --> 05:06:30,080 ANIMAL MODELS AND HOPEFULLY 7086 05:06:30,080 --> 05:06:32,040 HUMANS. AGAIN, IT IS THE IDEA OF 7087 05:06:32,040 --> 05:06:37,640 GROWING THE CANCER CELLS, TUMOR 7088 05:06:37,640 --> 05:06:39,240 CELLS IN CONTEXT OF TISSUE. YOU 7089 05:06:39,240 --> 05:06:40,720 CAN MAKE IT AS COMPLICATED AS 7090 05:06:40,720 --> 05:06:42,600 YOU WANT, T-CELLS, MACROPHAGES 7091 05:06:42,600 --> 05:06:45,800 SO STEP BY STEP WE ARE BUILDING 7092 05:06:45,800 --> 05:06:47,280 IN THIS PHYSIOLOGICAL COMPLEXITY 7093 05:06:47,280 --> 05:06:50,160 MANY THE MODELS. FOR THAT, THE 7094 05:06:50,160 --> 05:06:51,640 ENGINEERING APPROACH IS VERY 7095 05:06:51,640 --> 05:06:53,880 GOOD. WE PUBLISH THIS PAPER 7096 05:06:53,880 --> 05:06:59,280 COUPLE OF YEARS AGO, THIS WAS 7097 05:06:59,280 --> 05:07:01,080 CANCER CELL CARCINOMA, WE 7098 05:07:01,080 --> 05:07:03,320 STARTED WITH THE FIRE BREATHING 7099 05:07:03,320 --> 05:07:05,960 PROTOCOLS, WE ADDED SEC CELLS, 7100 05:07:05,960 --> 05:07:07,520 IN THIS CASE LABELED. WITH 7101 05:07:07,520 --> 05:07:08,440 FLUORESCENCE AND YOU CAN SEE 7102 05:07:08,440 --> 05:07:10,440 THAT WE CAN SEE THE CELLS 7103 05:07:10,440 --> 05:07:14,000 GROWING IN THE TISSUE. WITH 7104 05:07:14,000 --> 05:07:15,520 HISTOLOGY, VALIDATED BY 7105 05:07:15,520 --> 05:07:18,320 ALTERNATIVE ASSAY METHOD. THEN 7106 05:07:18,320 --> 05:07:20,800 WE DEVELOP ASSAY, CANCER CELLS 7107 05:07:20,800 --> 05:07:25,520 WERE LABEL WITH GFP OR RFP, RED 7108 05:07:25,520 --> 05:07:26,800 FLUORESCENT PROTEIN. YOU CAN SEE 7109 05:07:26,800 --> 05:07:30,240 AFTER TWO DAYS, YOUR SILL YOU 7110 05:07:30,240 --> 05:07:34,920 CAN SEE REDUCTION OF THE TUMORS 7111 05:07:34,920 --> 05:07:37,040 AND YOUR SILL IS USE #-D IN 7112 05:07:37,040 --> 05:07:39,000 CLINIC FOR SEC. WE WENT AHEAD 7113 05:07:39,000 --> 05:07:42,760 AND DEVELOPED THE ASSAY IN A 96 7114 05:07:42,760 --> 05:07:45,440 WELL PLATE AND THIS IS MY SLIDE 7115 05:07:45,440 --> 05:07:46,920 TO SHOW YOU CAN MAKE THIS ASSAY 7116 05:07:46,920 --> 05:07:51,000 IN A 96 WELL PLATE, NOT -- Z 7117 05:07:51,000 --> 05:07:54,080 FACTORS ARE .66 AND FOR THE 7118 05:07:54,080 --> 05:07:58,160 GREEN WHICH IS THE FIBROBLAST 7119 05:07:58,160 --> 05:08:00,040 LABEL GREEN SO THAT WAS OUR 7120 05:08:00,040 --> 05:08:01,120 INTERNAL CANCER SCREEN SO WE HAD 7121 05:08:01,120 --> 05:08:05,160 THE CANCER CELLS IN RED AND 7122 05:08:05,160 --> 05:08:06,160 FIBROBLAST IN GREEN SO KILL 7123 05:08:06,160 --> 05:08:07,880 CANCER CELLS BUT NOT FIBROBLAST 7124 05:08:07,880 --> 05:08:09,160 SO WESTERN ABLE TO HAVE A GOOD 7125 05:08:09,160 --> 05:08:14,400 ASSAY WINDOW FOR THE FIBROBLAST 7126 05:08:14,400 --> 05:08:18,000 AND .36 DOABLE ASSAY SO NOT BAD 7127 05:08:18,000 --> 05:08:20,960 FOR A BIOPRINT TISSUE MODEL WITH 7128 05:08:20,960 --> 05:08:23,480 CANCER GROWING IN THEM. THEN WE 7129 05:08:23,480 --> 05:08:25,160 DID PROOF OF CONCEPT WITH COUPLE 7130 05:08:25,160 --> 05:08:32,120 OF CONTROLS, EURO SILL AND CARBO 7131 05:08:32,120 --> 05:08:33,680 PLATIN. WE TAKE THE SAME CANCER 7132 05:08:33,680 --> 05:08:37,800 CELLS IN 3D AND TRADITIONAL 2D 7133 05:08:37,800 --> 05:08:41,640 MODEL. WHAT WAS INTERESTING IS 7134 05:08:41,640 --> 05:08:43,280 THE FIVE NEUROUROSILL IN 2D 7135 05:08:43,280 --> 05:08:46,560 DIDN'T HAVE ANY EFFECT IN CELL 7136 05:08:46,560 --> 05:08:47,880 PROLIFERATION BUT PUT IN CONTEXT 7137 05:08:47,880 --> 05:08:51,520 OF 3D IT HAD AN EFFECT. SO THAT 7138 05:08:51,520 --> 05:08:52,840 WAS VERY INTERESTING FOR US 7139 05:08:52,840 --> 05:08:56,520 BECAUSE IT SAYS THAT IN 3D YOU 7140 05:08:56,520 --> 05:08:57,680 CAPTURE BIOLOGY THAT YOU MIGHT 7141 05:08:57,680 --> 05:09:00,120 NOT CAPTURE IN A 2D ASSAY WHERE 7142 05:09:00,120 --> 05:09:02,600 CELLS ARE PROLIFERATING. BUT 7143 05:09:02,600 --> 05:09:04,280 COMPOUNDS WERE THE SAME, DIDN'T 7144 05:09:04,280 --> 05:09:05,720 MATTER. SO THESE DIFFERENCES IN 7145 05:09:05,720 --> 05:09:06,800 ASSESSING WHAT WORKS AND WHAT 7146 05:09:06,800 --> 05:09:08,280 DOESN'T WORK AND WHAT TYPE OF 7147 05:09:08,280 --> 05:09:12,360 ASSAY, WHERE WE ARE. THIS IS A 7148 05:09:12,360 --> 05:09:14,360 STATUS WHERE WE ARE NOW WITH THE 7149 05:09:14,360 --> 05:09:17,240 3D MODELS SO DEVELOP PROTOCOLS 7150 05:09:17,240 --> 05:09:20,400 TO MAKE VASCULATURE. BY MIXING 7151 05:09:20,400 --> 05:09:22,720 ENDOTHELIAL CELLS, FIBROBLASTS 7152 05:09:22,720 --> 05:09:26,720 AND PERICYTESS. WE CAN USE 7153 05:09:26,720 --> 05:09:29,920 BIOPRINTING TO CREATE PATTERNS 7154 05:09:29,920 --> 05:09:31,440 THIS ZIG ZAG PATTERN, WHY DOING 7155 05:09:31,440 --> 05:09:33,720 THAT? NOT BIOLOGICALLY RELEVANT 7156 05:09:33,720 --> 05:09:35,400 BUT IT IS ACTUALLY A VERY GOOD 7157 05:09:35,400 --> 05:09:37,400 PATTERN BECAUSE IT ALLOWS US TO 7158 05:09:37,400 --> 05:09:42,120 USE HIGH CONTENT IMAGING TO 7159 05:09:42,120 --> 05:09:47,160 READILY ASSESS AND QUANTITATE 7160 05:09:47,160 --> 05:09:48,320 VASCULAR FORMATION OF BIOGENESIS 7161 05:09:48,320 --> 05:09:50,080 SO BY CREATING THE EMPTY SPACES 7162 05:09:50,080 --> 05:09:53,320 WITH A HYDRO GEL YOU CAN SEE 7163 05:09:53,320 --> 05:09:56,200 VESSELS FORMING IN THIS SPACES 7164 05:09:56,200 --> 05:09:57,840 THEN WE CAN GO EASILY WITH THE 7165 05:09:57,840 --> 05:09:59,840 MASKS OR THE HIGH CONTENT READER 7166 05:09:59,840 --> 05:10:01,520 AND ASSESS HOW MUCH VASCULATURE 7167 05:10:01,520 --> 05:10:03,880 WE HAVE IN THESE SPACES. WE CALL 7168 05:10:03,880 --> 05:10:08,160 THAT ANGIOGENESIS. SO ABLE TO 7169 05:10:08,160 --> 05:10:10,480 DO VASSALRIESED SKIN, HAVE A LOT 7170 05:10:10,480 --> 05:10:12,080 OF CONTENT IMAGING LOOK AT THE 7171 05:10:12,080 --> 05:10:15,720 DIFFERENT LAYERS OF THE SKIN 7172 05:10:15,720 --> 05:10:17,160 IMMUNOHISTOCHEMISTRY. SO THIS 7173 05:10:17,160 --> 05:10:20,080 MODEL WE DECIDED TO REPRODUCE A 7174 05:10:20,080 --> 05:10:23,240 MODEL OF ECTOPIC DERMATITIS. 7175 05:10:23,240 --> 05:10:25,520 IMMUNE DISEASE, IT IS TRIGGERED 7176 05:10:25,520 --> 05:10:29,360 BY SOME INSULT, THROUGH THE SKIN 7177 05:10:29,360 --> 05:10:32,720 AND THAT GETS ACTIVATES TH 2 7178 05:10:32,720 --> 05:10:35,160 CELLS THAT TWO BACK TO THE SKIN, 7179 05:10:35,160 --> 05:10:41,480 AND THEN THIS IL 4 AND IL 13 THE 7180 05:10:41,480 --> 05:10:43,360 OTHER THING WE ARE LEARNING IS 7181 05:10:43,360 --> 05:10:44,960 DIFFICULT TO REPRODUCE DISEASES 7182 05:10:44,960 --> 05:10:48,280 END TO END IN THESE MODELS 7183 05:10:48,280 --> 05:10:50,880 BECAUSE WE HAD SKIN BUT NO LYMPH 7184 05:10:50,880 --> 05:10:52,240 NODE SO WE WOULDN'T BE ABLE TO 7185 05:10:52,240 --> 05:10:53,400 REPRODUCE THE WHOLE DISEASE END 7186 05:10:53,400 --> 05:10:57,040 TO END. WHAT WE DECIDED TO DO IS 7187 05:10:57,040 --> 05:11:01,280 LOOK LET'S START SIMPLE, WE WILL 7188 05:11:01,280 --> 05:11:03,240 USE IL 4 TO ACTIVATE THE DISEASE 7189 05:11:03,240 --> 05:11:08,280 PHENOTYPE. THIS WAS PUBLISHED 7190 05:11:08,280 --> 05:11:10,920 LIKE AGAIN COUPLE OF YEARS AGO, 7191 05:11:10,920 --> 05:11:13,160 SO WE CREATED THE VASCULARIZE 7192 05:11:13,160 --> 05:11:15,680 SKIN AND HAVE NORMAL AND THEN 7193 05:11:15,680 --> 05:11:16,840 INDUCE DISEASE PHENOTYPE WE 7194 05:11:16,840 --> 05:11:22,960 ADDED IL 4. PARTICULAR PROTOCOL 7195 05:11:22,960 --> 05:11:24,280 ADDED THE SAME DAY TISSUE WAS 7196 05:11:24,280 --> 05:11:26,200 ADDED TO THE INTERFACE. YOU CAN 7197 05:11:26,200 --> 05:11:28,480 GO BACK AND SEE BY HISTOLOGY, 7198 05:11:28,480 --> 05:11:32,560 THIS IS A NORMAL AND THIS IS THE 7199 05:11:32,560 --> 05:11:36,000 ECTOPIC DERMATITIS BY ADDING IL 7200 05:11:36,000 --> 05:11:41,800 4 YOU USE HYPO DISRUPTED 7201 05:11:41,800 --> 05:11:42,720 DIFFERENTIATION OF THE 7202 05:11:42,720 --> 05:11:44,800 KERATINOCYTES IN THE EPIDERMIS 7203 05:11:44,800 --> 05:11:46,560 THEN LOOK AT MARKERS OF 7204 05:11:46,560 --> 05:11:47,200 DIFFERENT DIFFERENTIATIONS SO 7205 05:11:47,200 --> 05:11:50,920 YOU CAN SEE LOWER IS WIN OF 7206 05:11:50,920 --> 05:11:54,320 THEM, NORMAL YOU SEE HIGH, IN 7207 05:11:54,320 --> 05:11:56,560 ECTOPIC DERMATITIS DISAPPEARS, 7208 05:11:56,560 --> 05:11:57,880 SAME FOR E CAD HEREIN. THE 7209 05:11:57,880 --> 05:12:01,960 QUESTION IS LOOKS BY HISTOLOGY 7210 05:12:01,960 --> 05:12:08,960 WE HAVE A GOOD MODEL FOR ECZEMA 7211 05:12:08,960 --> 05:12:10,960 PROOF OF CONCEPT, IL 2 SIGNAL 7212 05:12:10,960 --> 05:12:13,680 JAK STAT PATHWAY SO LET'S ADD 7213 05:12:13,680 --> 05:12:16,640 JACK INHIBITORS TO SEE WE CAN 7214 05:12:16,640 --> 05:12:21,920 PREVENT THESE DISEASE PHENOTYPE 7215 05:12:21,920 --> 05:12:25,040 HERE SHOWING DIFFERENT JAK 7216 05:12:25,040 --> 05:12:27,760 INHIBITORS WE ALSO TRIED A 7217 05:12:27,760 --> 05:12:29,560 IMMUNOSUPPRESSER. SO THE JACK 7218 05:12:29,560 --> 05:12:30,920 INHIBITORS CORRECTED THE DISEASE 7219 05:12:30,920 --> 05:12:34,600 PHENOTYPE, NORMAL,K ECZEMA AND 7220 05:12:34,600 --> 05:12:36,320 JAK 1, 2 AND 3 INHIBITORS AND 7221 05:12:36,320 --> 05:12:37,320 THEY CORRECT THE DISEASE 7222 05:12:37,320 --> 05:12:41,000 PHENOTYPE BY HISTOLOGY. WE 7223 05:12:41,000 --> 05:12:43,320 DIDN'T HAVE ANY P CELLS, THAT IS 7224 05:12:43,320 --> 05:12:45,720 THE TARGET OF CELLS SO MAKES 7225 05:12:45,720 --> 05:12:47,320 SENSE MAKES IT EVEN WORK IN THIS 7226 05:12:47,320 --> 05:12:49,240 ASSAY. THIS GOES ABOUT YOU ALSO 7227 05:12:49,240 --> 05:12:52,880 UNDERSTANDING WHAT IS THE 7228 05:12:52,880 --> 05:12:56,240 BIOLOGY OF ETIOLOGY YOUR ASSAY 7229 05:12:56,240 --> 05:12:58,080 CAPTURES? YOU CAPTURE WOULD MAKE 7230 05:12:58,080 --> 05:12:59,360 SENSE AND CAPTURE WHAT DOESN'T 7231 05:12:59,360 --> 05:13:01,320 MAKE SENSE AND THIS IS 7232 05:13:01,320 --> 05:13:03,760 DEMONSTRATED HERE. WE LIKE 7233 05:13:03,760 --> 05:13:06,320 ELECTRICAL RESISTANCE, IN 96 7234 05:13:06,320 --> 05:13:09,080 WELL PLATE, THE NORMAL IS HIGH, 7235 05:13:09,080 --> 05:13:13,400 THE ECZEMA LOW AND THEN J,K 7236 05:13:13,400 --> 05:13:14,560 INHIBITORS WERE CORRECT EVEN AND 7237 05:13:14,560 --> 05:13:16,120 CONFIRM WITH ADDITIONAL LIKE 7238 05:13:16,120 --> 05:13:18,440 MARKERS OF DIFFERENTIATION. SO 7239 05:13:18,440 --> 05:13:20,400 AGAIN VALIDATION PROOF OF 7240 05:13:20,400 --> 05:13:22,520 CONCEPT WITH SOME COMPOUND WE 7241 05:13:22,520 --> 05:13:24,240 THINK SHOULD WORK AND SOME 7242 05:13:24,240 --> 05:13:25,080 COMPOUNDS WE THINK SHOULDN'T 7243 05:13:25,080 --> 05:13:28,840 WORK. WHAT WAS INTERESTING IS 7244 05:13:28,840 --> 05:13:34,360 THAT WE WENT BACK TO WE HAVE ALL 7245 05:13:34,360 --> 05:13:37,720 THESE DIFFERENT MODELS WE HAVE 7246 05:13:37,720 --> 05:13:39,400 VASCULARIZED SKIN, DOESN'T MAKE 7247 05:13:39,400 --> 05:13:40,160 A DIFFERENCE. 7248 05:13:40,160 --> 05:13:41,960 DO WE HAVE TO DO A FULL 7249 05:13:41,960 --> 05:13:43,760 THICKNESS VASCULARIZED SKIN FOR 7250 05:13:43,760 --> 05:13:45,400 ECZEMA? THE TRUTH IS THAT WHAT 7251 05:13:45,400 --> 05:13:47,000 WAS REALLY INTERESTING IS THAT 7252 05:13:47,000 --> 05:13:49,120 IF YOU LOOK AT ELECTRICAL 7253 05:13:49,120 --> 05:13:51,520 RESISTANCE, AND LOOK JUST AT THE 7254 05:13:51,520 --> 05:13:55,760 DERMIS, BY ADDING THE IL 4 IT 7255 05:13:55,760 --> 05:13:59,000 DOESN'T HAVE EFFECT ON TIER 7256 05:13:59,000 --> 05:14:01,280 VALUE FOR JUST EPIDERMIS. THE 7257 05:14:01,280 --> 05:14:04,320 JAK INHIBITOR ESSENTIALLY KILL 7258 05:14:04,320 --> 05:14:06,360 THE EPIDERMIS, THAT WAS 7259 05:14:06,360 --> 05:14:11,440 INTERESTING AND BETWEEN THE 7260 05:14:11,440 --> 05:14:12,560 NON-VASCULARIZED AND 7261 05:14:12,560 --> 05:14:13,480 VASCULARIZED DIDN'T MAKE A 7262 05:14:13,480 --> 05:14:16,600 DIFFERENCE FOR THE READ OUT. SO 7263 05:14:16,600 --> 05:14:18,200 DON'T NEED VASCULATURE IF I WANT 7264 05:14:18,200 --> 05:14:21,120 TO LOOK AT THIS TIER EFFECT ON 7265 05:14:21,120 --> 05:14:22,800 THE ECZEMA MODEL. WHEN YOU LOOK 7266 05:14:22,800 --> 05:14:26,280 AT ONE OF THE CYTOKINES, SO WE 7267 05:14:26,280 --> 05:14:27,640 ALSO TOOK SUPERNATANTS AND 7268 05:14:27,640 --> 05:14:29,480 DIFFERENT CYTOKINES SO ATTACK 7269 05:14:29,480 --> 05:14:31,840 SIN IS ONE OF THE CYTOKINES AS A 7270 05:14:31,840 --> 05:14:33,320 BIOMARKER FOR ECZEMA. WHAT IS 7271 05:14:33,320 --> 05:14:34,880 INTERESTING IF YOU TAKE 7272 05:14:34,880 --> 05:14:36,680 KERATINOCYTES AND FIBROBLASTS 7273 05:14:36,680 --> 05:14:39,120 AND EVEN EPIDERMIS, 7274 05:14:39,120 --> 05:14:41,360 DIFFERENTIATED AND STIMULATE 7275 05:14:41,360 --> 05:14:43,520 WITH IL 4, YOU SEE NOTHING. 7276 05:14:43,520 --> 05:14:47,080 THERE IS NO SECRETION OF TOXIN. 7277 05:14:47,080 --> 05:14:49,120 ONLY WHEN YOU START WITH THE 7278 05:14:49,120 --> 05:14:51,240 FULL SIGNALS THAT INCLUDE 7279 05:14:51,240 --> 05:14:52,400 FIBROBLASTS LAYER, THAT'S WHEN 7280 05:14:52,400 --> 05:14:56,600 YOU START SEEING SECRETION OF AO 7281 05:14:56,600 --> 05:14:58,720 TAXIN AND REVERSE WITH JAK 7282 05:14:58,720 --> 05:15:01,880 INHIBITORS SO FOR US THIS WAS AN 7283 05:15:01,880 --> 05:15:06,560 EXAMPLE OF LIKE HITTING THE 7284 05:15:06,560 --> 05:15:07,480 SWEET SPOT FOR THAT PARTICULAR 7285 05:15:07,480 --> 05:15:09,080 READ OUT AND THAT PARTICULAR 7286 05:15:09,080 --> 05:15:10,560 EFFECT AND DISEASE PHENOTYPE. 7287 05:15:10,560 --> 05:15:13,120 YOU COULDN'T DO THAT IN 2D. NOW 7288 05:15:13,120 --> 05:15:15,200 WE HAVE OTHER EXAMPLES WE SEE 7289 05:15:15,200 --> 05:15:16,240 SIMILAR EFFECTS THAT ONLY WHEN 7290 05:15:16,240 --> 05:15:19,960 YOU HIT THE RIGHT PHYSIOLOGICAL 7291 05:15:19,960 --> 05:15:21,120 COMPLEXITY YOU SEE A PHENOTYPE 7292 05:15:21,120 --> 05:15:23,480 THAT IS RELEVANT. CLINICALLY 7293 05:15:23,480 --> 05:15:30,480 RELEVANT. GOING TO GIVE AN 7294 05:15:30,480 --> 05:15:31,400 EXAMPLE OF ANOTHER MODEL WE 7295 05:15:31,400 --> 05:15:37,080 DEVELOPED. THIS IS NEUROVASCULAR 7296 05:15:37,080 --> 05:15:39,640 UNIT. ANOTHER VERY NICE THING 7297 05:15:39,640 --> 05:15:42,280 METHOD WE USE IS THAT YOU CAN 7298 05:15:42,280 --> 05:15:46,160 REPURPOSE THE PROTOCOLS FOR 7299 05:15:46,160 --> 05:15:48,720 DIFFERENT TISSUE SO THE PROTOCOL 7300 05:15:48,720 --> 05:15:52,360 FOR VASCULATURE IS REDUNDANCY IN 7301 05:15:52,360 --> 05:15:55,280 NATURE. ENDOTHELIAL CELLS, 7302 05:15:55,280 --> 05:15:57,560 FIBROBLASTS OR IN THE BRAIN 7303 05:15:57,560 --> 05:15:59,600 ASTROCYTES TO FORM VASCULATURE 7304 05:15:59,600 --> 05:16:02,920 NETWORKS SO YOU CAN USE THE SAME 7305 05:16:02,920 --> 05:16:07,440 BASIC PROTOCOL AND USING BRAIN 7306 05:16:07,440 --> 05:16:11,360 ENENDOTHELIAL CELLS AND BRAIN 7307 05:16:11,360 --> 05:16:14,920 ASTROCYTES AND MIX THAT WITH THE 7308 05:16:14,920 --> 05:16:16,560 SAME HYDRA GELS, GELATIN BASED 7309 05:16:16,560 --> 05:16:18,360 FIBRIN BASED HYDRA JELL AND 7310 05:16:18,360 --> 05:16:20,680 CREATE PATTERNS AND FORM 7311 05:16:20,680 --> 05:16:21,480 VASCULATURE SO IN THIS 7312 05:16:21,480 --> 05:16:24,880 PARTICULAR ASSAY, AGAIN WE 7313 05:16:24,880 --> 05:16:28,000 CREATED THIS RING DONUT RATHER 7314 05:16:28,000 --> 05:16:29,680 AND THE PURPOSE IS SO YOU CAN 7315 05:16:29,680 --> 05:16:31,800 MEASURE READILY VASCULAR GENESIS 7316 05:16:31,800 --> 05:16:33,520 AN ANGIOGENESIS IN THE MODEL. 7317 05:16:33,520 --> 05:16:34,960 NOT FOR OTHER PURPOSE, VERY 7318 05:16:34,960 --> 05:16:36,640 PRACTICAL BUT ESSENTIALLY WHAT 7319 05:16:36,640 --> 05:16:39,440 YOU DO IS YOU BITE DONUTS IN 7320 05:16:39,440 --> 05:16:42,160 EACH WELL AN COVER WITH EMPTY 7321 05:16:42,160 --> 05:16:44,760 HYDRA GEL. WE WAIT ESSENTIALLY 7322 05:16:44,760 --> 05:16:48,080 FOR 21 DAYS AND YOU CAN SEE THIS 7323 05:16:48,080 --> 05:16:49,560 BEAUTIFUL VASCULATURE, THIS IS A 7324 05:16:49,560 --> 05:16:52,320 CARTOON, JUST BEFORE WE SHOW YOU 7325 05:16:52,320 --> 05:16:54,120 THAT FORMATION OF VASCULATURE, 7326 05:16:54,120 --> 05:16:57,160 TO GIVE YOU A SENSE THE 7327 05:16:57,160 --> 05:16:59,000 PREDICTABILITY IS REPRODUCIBLE 7328 05:16:59,000 --> 05:17:00,800 FROM WELL TO WELL TO WELL, WE 7329 05:17:00,800 --> 05:17:04,520 HAVE GOOD CV ON THE AREA, OF THE 7330 05:17:04,520 --> 05:17:08,960 DONUT AND THE ROUNDEDNESS, 7331 05:17:08,960 --> 05:17:19,480 PRETTY REPRODUCIBLE PROCESS. IF 7332 05:17:40,680 --> 05:17:43,480 YOU WAIT LONG ENOUGH RIGHT 7333 05:17:43,480 --> 05:17:45,680 CONDITIONS YOU CAN USE HIGH 7334 05:17:45,680 --> 05:17:48,400 CONTENT IMAGING CONFOCAL ENOUGH 7335 05:17:48,400 --> 05:17:51,920 EVEN THESE TISSUES ARE 200 7336 05:17:51,920 --> 05:17:54,720 MICROTWO PHOTON CLEARING YOU CAN 7337 05:17:54,720 --> 05:17:57,000 ACTUALLY STICK IN THIS CASE WE 7338 05:17:57,000 --> 05:17:59,360 HAVE A PHOENIX SO YOU CAN 7339 05:17:59,360 --> 05:18:01,640 MEASURE REAL TIME LIKE FORMATION 7340 05:18:01,640 --> 05:18:03,600 OF VASCULATURE NETWORK AND YOU 7341 05:18:03,600 --> 05:18:06,920 CAN QUANTITATE THAT CLEARLY WITH 7342 05:18:06,920 --> 05:18:07,840 ALGORITHMS FROM THE INSTRUMENT 7343 05:18:07,840 --> 05:18:09,560 AND YOU CAN SEE FORMATION OF THE 7344 05:18:09,560 --> 05:18:11,160 VESSELS AND YOU CAN QUANTITATE 7345 05:18:11,160 --> 05:18:19,720 FORMATION OF THE VESSELS. WE 7346 05:18:19,720 --> 05:18:21,840 HAVE NORMAL, WHAT CAN WE DO WITH 7347 05:18:21,840 --> 05:18:25,080 THIS. TO INTRODUCE A GLIOMA CELL 7348 05:18:25,080 --> 05:18:28,080 TO CREATE A MODEL OF GLIOMA IN 7349 05:18:28,080 --> 05:18:29,960 CONTEXT OF NEUROVAS QUEUE 7350 05:18:29,960 --> 05:18:31,240 ALREADY A UNIT. IT IS NOT 7351 05:18:31,240 --> 05:18:34,920 PERFECT, DOESN'T HAVE NEURONS. 7352 05:18:34,920 --> 05:18:36,920 THAT IS THE NEXT STEP BUT TRYING 7353 05:18:36,920 --> 05:18:39,280 TO SHOW THE PROCESS IS VERSATILE 7354 05:18:39,280 --> 05:18:42,480 AND MODULAR YOU CAN CHOOSE WHAT 7355 05:18:42,480 --> 05:18:46,960 CELLS YOU WANT TO INCLUDE AND 7356 05:18:46,960 --> 05:18:50,200 SEE, SO IN THIS CASE WE WANT TO 7357 05:18:50,200 --> 05:18:55,560 CANCER CELLS GROWING IN THE 7358 05:18:55,560 --> 05:18:58,040 RELEVANT TISSUE VASCULATURE. WE 7359 05:18:58,040 --> 05:18:59,880 ADDED A GLIOMA CELL IN THE 7360 05:18:59,880 --> 05:19:03,520 MIDDLE OF DONUT THEN WE LOOK AT 7361 05:19:03,520 --> 05:19:07,840 FORMATION OF VASCULATURE. 7362 05:19:07,840 --> 05:19:09,600 FORMATION IN THIS CASE 7363 05:19:09,600 --> 05:19:12,920 ENDOTHELIAL CELLS LABEL WITH 7364 05:19:12,920 --> 05:19:16,440 GFP. THE PATIENT DERIVED GLIOMA 7365 05:19:16,440 --> 05:19:18,360 CELLS LABELED WITH CHERRY SO WE 7366 05:19:18,360 --> 05:19:23,440 CAN USE IMAGING TO LOOK AT THE 7367 05:19:23,440 --> 05:19:25,040 EFFECTS, GROWTH OF THE VESSELS 7368 05:19:25,040 --> 05:19:28,880 AND GROWTH OF THE TUMOR. THEN 7369 05:19:28,880 --> 05:19:31,040 YOU CAN PLAY GAMES AND SAY WHAT 7370 05:19:31,040 --> 05:19:32,160 DIFFERENCE -- DOES THE 7371 05:19:32,160 --> 05:19:33,000 VASCULATURE MAKE A DIFFERENCE IN 7372 05:19:33,000 --> 05:19:35,400 THE GROWTH OF TUMOR? DOES TUMOR 7373 05:19:35,400 --> 05:19:36,720 MAKE A DIFFERENCE ON THE GROWTH 7374 05:19:36,720 --> 05:19:39,240 OF THE VESSEL, SO THESE ARE 7375 05:19:39,240 --> 05:19:40,120 INTERESTING QUESTIONS BECAUSE WE 7376 05:19:40,120 --> 05:19:45,520 ARE TRYING TO ASSESS WHETHER 7377 05:19:45,520 --> 05:19:46,240 THIS CROSS TALK BETWEEN VAS VIEW 7378 05:19:46,240 --> 05:19:48,880 LATURE AND TUMOR WHETHER THAT 7379 05:19:48,880 --> 05:19:52,360 AFFECTS EACH ONE OF THEM. THEN 7380 05:19:52,360 --> 05:19:56,720 ASSAY WORKSHOP. WE DID THAT IN A 7381 05:19:56,720 --> 05:19:59,280 96 WELL PLATE AND FOCUS SCREEN. 7382 05:19:59,280 --> 05:20:01,000 HOW MANY COMPOUNDS, THERE 7383 05:20:01,000 --> 05:20:01,960 WEREN'T TOO MANY LIKE 20 OR 7384 05:20:01,960 --> 05:20:06,880 SOMETHING. THE IDEA HERE WAS TO 7385 05:20:06,880 --> 05:20:12,400 THINK OF IT LIKE A XENOGRAPH. A 7386 05:20:12,400 --> 05:20:16,040 DAY 13 WE START SEEING TUMORS 7387 05:20:16,040 --> 05:20:18,200 GROWING AND THEN THIS IS ALL YOU 7388 05:20:18,200 --> 05:20:19,960 CAN DO REAL TIME MEASUREMENTS 7389 05:20:19,960 --> 05:20:23,080 AND YOU CAN MEASURE THE TUMOR 7390 05:20:23,080 --> 05:20:26,520 DAY 13 AND MEASURE TUMOR DAY 21, 7391 05:20:26,520 --> 05:20:28,880 LOOK AT DIFFERENCES IN GROWTH. 7392 05:20:28,880 --> 05:20:30,440 WITH OR WITHOUT COMPOUNDS. SO 7393 05:20:30,440 --> 05:20:33,440 HERE I SHOW YOU PLATE ON DAY 13 7394 05:20:33,440 --> 05:20:35,800 ON TOP AND PLATE MAP HERE WE 7395 05:20:35,800 --> 05:20:37,480 HAVE THE DMSO CONTROLS ON THE 7396 05:20:37,480 --> 05:20:39,080 ONE SIDE AND THEN WE HAVE FOUR 7397 05:20:39,080 --> 05:20:40,280 DOSES ON TOP AND FOUR ON THE 7398 05:20:40,280 --> 05:20:45,360 BOTTOM. OF EACH PLATE. WHAT YOU 7399 05:20:45,360 --> 05:20:49,240 CAN SEE EASILY IS DAY 21 AFTER 7400 05:20:49,240 --> 05:20:52,720 SEVEN DAYS OF BIDING THE 7401 05:20:52,720 --> 05:20:55,080 COMPOUND MOST VASCULATURE, THESE 7402 05:20:55,080 --> 05:20:57,240 COMPOUNDS WERE SELECTED 7403 05:20:57,240 --> 05:20:59,080 CHEMOTHERAPEUTICS MORE TARGETED 7404 05:20:59,080 --> 05:21:00,600 COMPOUNDS, MEC INHIBITORS, WE 7405 05:21:00,600 --> 05:21:02,480 WERE TRYING TO LOOK AT EFFECT OF 7406 05:21:02,480 --> 05:21:05,040 COMPOUNDS WE KNEW SORT OF ALSO 7407 05:21:05,040 --> 05:21:07,640 AFFECTED THE SORT OF 7408 05:21:07,640 --> 05:21:09,720 MICROENVIRONMENT TUMOR CELLS IN 7409 05:21:09,720 --> 05:21:11,080 THE MICROENVIRONMENT. AT THE END 7410 05:21:11,080 --> 05:21:13,080 OF THE DAY, MOST OF THE EFFECTS 7411 05:21:13,080 --> 05:21:15,440 WERE REALLY IF WE HAD AN EFFECT 7412 05:21:15,440 --> 05:21:18,200 ON THE TUMOR HAD EFFECT ON 7413 05:21:18,200 --> 05:21:20,120 VASCULATURE. WE WERE VERY EASILY 7414 05:21:20,120 --> 05:21:22,160 SEE WHAT COMPOUND IS FLUORESCE 7415 05:21:22,160 --> 05:21:23,960 SENT BECAUSE IT FROM SLIDES, SO 7416 05:21:23,960 --> 05:21:26,160 YOU CAN REMOVE THAT EASILY. THE 7417 05:21:26,160 --> 05:21:27,800 BOTTOM LINE IS THAT THE ASSAY 7418 05:21:27,800 --> 05:21:29,960 WAS QUITE ROBUST. WE DIDN'T GET 7419 05:21:29,960 --> 05:21:31,560 THE RESULTS WE WERE EXPECTING 7420 05:21:31,560 --> 05:21:32,600 BUT AGAIN ALL THOSE COMPOUNDS 7421 05:21:32,600 --> 05:21:35,600 WERE DEVELOPED IN 2D SO MAYBE 7422 05:21:35,600 --> 05:21:38,600 THAT IS WHAT WE EXPECT. MEMBER 7423 05:21:38,600 --> 05:21:41,840 WHAT WE OUGHT TO DO IS START 7424 05:21:41,840 --> 05:21:43,480 SCREENS WITH ASSAYS AND FIND NEW 7425 05:21:43,480 --> 05:21:44,920 TARGETS AND NEW BIOLOGY THAT 7426 05:21:44,920 --> 05:21:48,000 WORK IN 3D AND NOT NECESSARILY 7427 05:21:48,000 --> 05:21:51,240 IN 2D BUT HOPEFULLY THEN WILL BE 7428 05:21:51,240 --> 05:21:52,480 MORE RELEVANT WHEN WE GO INTO 7429 05:21:52,480 --> 05:21:55,360 ANIMAL CLINIC. ALL I WANT TO 7430 05:21:55,360 --> 05:21:59,800 SHOW YOU HERE IS GENERATING DOSE 7431 05:21:59,800 --> 05:22:01,080 RESPONSES, PROBABLY HAVE THAT, 7432 05:22:01,080 --> 05:22:02,680 WE WOULD LIKE TO YET BUT THIS IS 7433 05:22:02,680 --> 05:22:05,200 WHERE WE ARE GOING TO, CAN STILL 7434 05:22:05,200 --> 05:22:08,440 -- CAN WE ALREADY ASSESS 7435 05:22:08,440 --> 05:22:10,560 ACTIVITY POTENCY AND EFFICACY OF 7436 05:22:10,560 --> 05:22:13,720 COMPOUNDS USING BIOPRINTED 7437 05:22:13,720 --> 05:22:16,200 TISSUES. SO THIS IS MY LAST 7438 05:22:16,200 --> 05:22:20,040 SLIDE. WANT THE GIVE A TASTE OF 7439 05:22:20,040 --> 05:22:22,560 WHAT WE ARE DOING WHERE ARE WE 7440 05:22:22,560 --> 05:22:24,160 GOING THE QUESTIONS WE ARE 7441 05:22:24,160 --> 05:22:28,080 TRYING TO ASK. IT IS NOT JUST 7442 05:22:28,080 --> 05:22:29,640 PRINTING THE TISSUES. I THINK 7443 05:22:29,640 --> 05:22:32,120 OUR BIGGEST CHALLENGE IS GETTING 7444 05:22:32,120 --> 05:22:35,560 THE RIGHT CELLS. AS I MENTION TO 7445 05:22:35,560 --> 05:22:37,400 MAKE THESE TISSUES MORE 7446 05:22:37,400 --> 05:22:38,800 PHYSIOLOGICALLY RELEVANT WE USE 7447 05:22:38,800 --> 05:22:40,600 PRIMARY CELLS AND IPS DERIVED 7448 05:22:40,600 --> 05:22:45,480 CELLS. THERE IS NO CLEAR WINNER. 7449 05:22:45,480 --> 05:22:47,080 IF YOU CAN YOU WANT TO USE 7450 05:22:47,080 --> 05:22:49,320 PRIMARY CELLS, FROM LOT TO LOT 7451 05:22:49,320 --> 05:22:50,560 SOMETIMES WORK, SOMETIMES IT 7452 05:22:50,560 --> 05:22:52,360 DOESN'T WORK. SOMETIMES THE 7453 05:22:52,360 --> 05:22:54,960 CELLSES LOOK OKAY BUT NO 7454 05:22:54,960 --> 05:22:56,800 VASCULATURE. WE DON'T KNOW WHY. 7455 05:22:56,800 --> 05:23:02,440 IPS DERIVED CELLS THEY ARE 7456 05:23:02,440 --> 05:23:05,520 EXPENSIVE. EITHER MAKE THEM IN 7457 05:23:05,520 --> 05:23:07,160 HOUSE OR BUY THEM ALREADY 7458 05:23:07,160 --> 05:23:09,760 DIFFERENTIATED. SO THE PROCESS 7459 05:23:09,760 --> 05:23:13,200 OF SCALING UP BATCH TO BATCH IS 7460 05:23:13,200 --> 05:23:15,040 BIG ISSUES FOR US AND A BIG 7461 05:23:15,040 --> 05:23:18,720 ISSUE FOR THE FIELD. 7462 05:23:18,720 --> 05:23:20,560 BIOPRINTING THE HYDRA GELS, I 7463 05:23:20,560 --> 05:23:24,040 THINK THAT IS SOMETHING WE ARE 7464 05:23:24,040 --> 05:23:26,680 WORKING ON. IT IS NOT BIG 7465 05:23:26,680 --> 05:23:28,360 HURDLE. TUSH SHOE VALIDATION IS 7466 05:23:28,360 --> 05:23:31,400 REALLY CRITICAL AS WELL, WHETHER 7467 05:23:31,400 --> 05:23:34,560 YOU USE IMMUNOHISTOCHEMISTRY, 7468 05:23:34,560 --> 05:23:36,120 SINGLE CELL SEQUENCING. WE ARE 7469 05:23:36,120 --> 05:23:38,760 LEARNING A LOT OF PLASTICITY IN 7470 05:23:38,760 --> 05:23:43,160 THE CELLS. AND ENDOTHELIAL CELLS 7471 05:23:43,160 --> 05:23:44,320 IS VERY DIFFERENT FROM 7472 05:23:44,320 --> 05:23:46,200 ENDOTHELIAL CELL FORMING TUBES 7473 05:23:46,200 --> 05:23:48,040 AND FORMING VASCULATURE. 7474 05:23:48,040 --> 05:23:48,960 UNDERSTANDING THESE DIFFERENCES 7475 05:23:48,960 --> 05:23:52,480 I THINK IS VERY IMPORTANT AND 7476 05:23:52,480 --> 05:23:54,640 BENCHMARKING THESE CHANGES TO 7477 05:23:54,640 --> 05:23:55,840 REAL CLINICAL SAMPLES I THINK 7478 05:23:55,840 --> 05:24:01,320 THAT IS VERY IMPORTANT AS WELL. 7479 05:24:01,320 --> 05:24:03,040 DOING SINGLE CELL SEQUENCING TO 7480 05:24:03,040 --> 05:24:04,480 UNDERSTAND PLASTICITY, 7481 05:24:04,480 --> 05:24:07,920 BENCHMARKING TISSUES. A LOT OF 7482 05:24:07,920 --> 05:24:08,640 IMMUNOHISTOCHEMISTRY WHEN WE CAN 7483 05:24:08,640 --> 05:24:13,400 PHARMACOLOGY. USE HIGH THROUGH 7484 05:24:13,400 --> 05:24:14,680 PUT SCREENING A LOT AND GENERATE 7485 05:24:14,680 --> 05:24:20,160 A LOT OF DATA. SO IT PERSON 7486 05:24:20,160 --> 05:24:21,640 BECOMES YOUR BEST FRIEND, THE 7487 05:24:21,640 --> 05:24:25,640 INFRASTRUCTURE YOU NEED TO STORE 7488 05:24:25,640 --> 05:24:31,680 AND ANALYZE THAT DATA, JUST 7489 05:24:31,680 --> 05:24:34,040 INCREDIBLE. IT IS A WHOLE POCKET 7490 05:24:34,040 --> 05:24:37,120 WE NEED TO SET UP TO REALLY BE 7491 05:24:37,120 --> 05:24:40,040 ABLE TO BE FUNCTIONAL DURING 7492 05:24:40,040 --> 05:24:41,640 THESE KINDS OF STUDIES. I THINK 7493 05:24:41,640 --> 05:24:42,600 THAT IS IT. THANK YOU VERY MUCH 7494 05:24:42,600 --> 05:24:49,560 [APPLAUSE] 7495 05:24:49,560 --> 05:24:52,240 >>THANK YOU SO MUCH, MARC. SO 7496 05:24:52,240 --> 05:24:53,520 COOL TO SEE YOU PRESENT. 7497 05:24:53,520 --> 05:25:03,880 QUESTIONS FOR MARC? 7498 05:25:06,680 --> 05:25:10,080 >>THAT WAS COOL. WHEN YOU ARE 7499 05:25:10,080 --> 05:25:11,720 SHOWING THE EXAMPLE, YOU CAN GO 7500 05:25:11,720 --> 05:25:13,840 NINE DAYS FOR THE BIOPRINTED 7501 05:25:13,840 --> 05:25:16,200 TISSUES AND THE ASSAY WAS 48 7502 05:25:16,200 --> 05:25:18,000 HOURS TWO DAYS FOR THE OTHER. SO 7503 05:25:18,000 --> 05:25:19,160 WONDERING ONE, COULD THAT 7504 05:25:19,160 --> 05:25:22,120 ACCOUNT FOR THE DIFFERENCE IN 7505 05:25:22,120 --> 05:25:23,720 ACTIVITIES FOR FIVE FU AND MAYBE 7506 05:25:23,720 --> 05:25:25,920 LEARN SOMETHING ABOUT THAT TOO. 7507 05:25:25,920 --> 05:25:26,720 ANY COMMENTS ON THAT? 7508 05:25:26,720 --> 05:25:30,600 >>ABSOLUTELY RIGHT. I THINK 7509 05:25:30,600 --> 05:25:32,560 MUCH CLOSER TO THE CLINIC AND 7510 05:25:32,560 --> 05:25:37,880 WHAT IS CLOSER TO ANIMAL MODELS 7511 05:25:37,880 --> 05:25:41,080 AND I THINK PROLIFERATION 7512 05:25:41,080 --> 05:25:45,760 ASSAYS, CELLS ARE GOING FAST, 7513 05:25:45,760 --> 05:25:47,920 THEY ARE A LOT OF MEDIA. THE 7514 05:25:47,920 --> 05:25:51,640 FACT THAT A COMPOUND DIDN'T WORK 7515 05:25:51,640 --> 05:25:54,960 FOR WHATEVER REASON, MEANS 7516 05:25:54,960 --> 05:25:56,520 (INAUDIBLE) MIGHT BE LONGER TIME 7517 05:25:56,520 --> 05:25:58,000 BUT IT IS JUST IN ONE WE DON'T 7518 05:25:58,000 --> 05:26:01,920 HAVE ANY EFFECT SIZE SO WE DON'T 7519 05:26:01,920 --> 05:26:03,080 KNOW THE RIGHT SIDE, THESE ARE 7520 05:26:03,080 --> 05:26:04,080 THE DIFFERENCES THEY ARE TRYING 7521 05:26:04,080 --> 05:26:06,080 TO ASSESS NOW. 7522 05:26:06,080 --> 05:26:08,040 >>THAT BRINGS UP A POINT I 7523 05:26:08,040 --> 05:26:09,720 DIDN'T REALLY CATCH. DO YOU 7524 05:26:09,720 --> 05:26:12,600 TREAT FOR NINE DAYS WITH 5 FU 7525 05:26:12,600 --> 05:26:14,160 FOR THAT NINE DAY EXPERIMENT? 7526 05:26:14,160 --> 05:26:17,640 >>SO WE TEND (INAUDIBLE) AND WE 7527 05:26:17,640 --> 05:26:18,720 -- (OVERLAPPING SPEAKERS) 7528 05:26:18,720 --> 05:26:21,320 >>OKAY. THANK YOU. SO HIGHER 7529 05:26:21,320 --> 05:26:21,920 DOSE DENSITY JUST TO -- 7530 05:26:21,920 --> 05:26:27,200 >>LONGER TIME. (INAUDIBLE) 7531 05:26:27,200 --> 05:26:29,240 >>DO YOU MIND TALKING INTO THE 7532 05:26:29,240 --> 05:26:29,560 MICROPHONE? 7533 05:26:29,560 --> 05:26:32,120 >>HI, MARC. I HAVE A QUESTION 7534 05:26:32,120 --> 05:26:39,200 AND IT MIGHT BE STUPID. THE 2D 7535 05:26:39,200 --> 05:26:40,800 SYSTEM YOU HAVE THE MATRIX IN 7536 05:26:40,800 --> 05:26:44,960 LIQUID FORM. AND 3D IN A GEL 7537 05:26:44,960 --> 05:26:46,880 FORM. MY UNDERSTANDING. MY 7538 05:26:46,880 --> 05:26:48,120 QUESTION IS WITH REGARDS TO WHEN 7539 05:26:48,120 --> 05:26:52,360 YOU ADD THE COMPOUND, DOES IS 7540 05:26:52,360 --> 05:26:53,520 SOLUBILITY OF COMPOUND CHANGE OR 7541 05:26:53,520 --> 05:26:56,320 HAVE YOU NOTICED ANY 7542 05:26:56,320 --> 05:26:59,400 VARIABILITY? 7543 05:26:59,400 --> 05:27:02,040 >>I GUESS WHEN WE SEE ACTIVITY 7544 05:27:02,040 --> 05:27:05,600 WE KNOW COMPOUND GETS IN. WHEN 7545 05:27:05,600 --> 05:27:07,640 WE DON'T SEE ACTIVITY WE HAVEN'T 7546 05:27:07,640 --> 05:27:10,320 DONE POSITIVE ANALYSIS OF 7547 05:27:10,320 --> 05:27:12,440 COMPOUND PENETRATION IN THESE 7548 05:27:12,440 --> 05:27:14,560 TISSUES. DYES GO IN THESE 7549 05:27:14,560 --> 05:27:16,040 BIOPRINTED TISSUES. IT IS 7550 05:27:16,040 --> 05:27:17,720 DIFFERENT FROM ORGANOIDS. AND 7551 05:27:17,720 --> 05:27:19,800 STEROIDS ORGANOIDS AN STEROIDS 7552 05:27:19,800 --> 05:27:21,960 ARE MORE DENSE AND THEN YOU HAVE 7553 05:27:21,960 --> 05:27:23,240 TIME, YOU HAVE A HARDER TIME 7554 05:27:23,240 --> 05:27:26,240 GETTING DYES INTO THE SYSTEM AND 7555 05:27:26,240 --> 05:27:29,680 PROBABLY COMPOUNDS TOO. THESE 7556 05:27:29,680 --> 05:27:32,200 TISSUES ARE NOT AS DENSE. SO I 7557 05:27:32,200 --> 05:27:33,800 THINK COMPOUNDS GO IN BUT WE 7558 05:27:33,800 --> 05:27:37,120 DON'T HAVE THE DATA TO SAY 7559 05:27:37,120 --> 05:27:38,120 POSITIVELY THAT IS WHAT HAPPENS. 7560 05:27:38,120 --> 05:27:40,920 THANK YOU. 7561 05:27:40,920 --> 05:27:44,240 >> 7562 05:27:44,240 --> 05:27:51,800 (OFF MIC) 7563 05:27:51,800 --> 05:27:54,600 VASCULATURE IN MY MIND IS BLOOD 7564 05:27:54,600 --> 05:27:56,280 VESSEL WITH ALLELE CELLS WITH A 7565 05:27:56,280 --> 05:27:58,680 CERTAIN ARCHITECTURE, WHEN YOU 7566 05:27:58,680 --> 05:28:00,520 ARE SHOWING THAT -- WHAT IS THE 7567 05:28:00,520 --> 05:28:02,480 NETWORK, WHAT IS THE NETWORK 7568 05:28:02,480 --> 05:28:12,800 LOOK LIKE? IS SOME CELLS LINED? 7569 05:28:12,800 --> 05:28:17,240 >>THE ONES YOU CAN FUND VAS 7570 05:28:17,240 --> 05:28:19,440 QUEUE LATURE, IT IS HOE LOW AND 7571 05:28:19,440 --> 05:28:21,640 WE CAN SEE THAT LITTLE 7572 05:28:21,640 --> 05:28:23,960 SECTIONING AND THAT DOES NOT PRO 7573 05:28:23,960 --> 05:28:25,960 FUSE. SO IN ORDER TO HAVE 7574 05:28:25,960 --> 05:28:27,520 PROFUSION YOU NEED TO GO TO A 7575 05:28:27,520 --> 05:28:30,920 DIFFERENT PLATFORM. MORE 7576 05:28:30,920 --> 05:28:32,520 MICROFLUIDIC TYPE OF PLATFORM. 7577 05:28:32,520 --> 05:28:35,720 BUT TO ANSWER YOUR QUESTION, 7578 05:28:35,720 --> 05:28:38,400 THESE ARE REAL VESSELS IN THE 7579 05:28:38,400 --> 05:28:40,080 SENSE -- YOU CAN SEE WHEN YOU 7580 05:28:40,080 --> 05:28:41,600 SECTION YOU CAN SEE ACTUALLY 7581 05:28:41,600 --> 05:28:41,800 THAT. 7582 05:28:41,800 --> 05:28:46,520 >>PRETTY COOL. 7583 05:28:46,520 --> 05:28:50,000 >>ANY MORE QUESTIONS? 7584 05:28:50,000 --> 05:28:51,640 >>THANK YOU SO MUCH, MARC. 7585 05:28:51,640 --> 05:29:01,800 [APPLAUSE] 7586 05:29:04,640 --> 05:29:06,360 WE HAVE TWO WORKSHOPS TALKING 7587 05:29:06,360 --> 05:29:08,600 ABOUT 3D MODELS. ONE ORGANIZED 7588 05:29:08,600 --> 05:29:13,480 WITH HELP OF DR. FERRER IN 7589 05:29:13,480 --> 05:29:14,560 COLLABORATION WITH BILL AND 7590 05:29:14,560 --> 05:29:17,600 MELINDA GATES FOUNDATION AND 3D 7591 05:29:17,600 --> 05:29:19,040 ANTI-DRUG DEVELOPMENT TWO FULL 7592 05:29:19,040 --> 05:29:21,040 DAYS OF 3D MODELS TALKS RECORDED 7593 05:29:21,040 --> 05:29:22,960 AVAILABLE FOR YOU TO VIEW. SO IF 7594 05:29:22,960 --> 05:29:25,040 YOU ARE INTERESTED IN THIS TYPE 7595 05:29:25,040 --> 05:29:26,840 OF RESEARCH I HIGHLY ENCOURAGE 7596 05:29:26,840 --> 05:29:30,040 YOU TO LOOK AT THOSE VIDEOS. WE 7597 05:29:30,040 --> 05:29:31,640 HAVE ANOTHER WORKSHOP ON CELL 7598 05:29:31,640 --> 05:29:35,720 BASED ASSAYS THAT HAD ONE DAY ON 7599 05:29:35,720 --> 05:29:36,880 COMPLEX VISUAL MODELS AS WELL. 7600 05:29:36,880 --> 05:29:38,960 SO ALL THESE RESOURCES ARE 7601 05:29:38,960 --> 05:29:42,800 AVAILABLE FOR YOU TO VIEW. ALL 7602 05:29:42,800 --> 05:29:49,760 RIGHT. OUR FINAL SPEAKER IS DR. 7603 05:29:49,760 --> 05:29:52,400 XIN XU, SENIOR SCIENTIST AND 7604 05:29:52,400 --> 05:29:55,360 DIRECTOR OF PHARMA KINETICS IN 7605 05:29:55,360 --> 05:29:56,360 THERAPEUTICS DEVELOPMENT BRANCH 7606 05:29:56,360 --> 05:29:57,880 DIVISION OF PRE-CLINICAL 7607 05:29:57,880 --> 05:30:01,040 INNOVATION AT NCATS. SHE IS ALSO 7608 05:30:01,040 --> 05:30:02,720 A LONG STANDING EDITORIAL BOARD 7609 05:30:02,720 --> 05:30:05,080 MEMBER OF THE AGM, WE ARE 7610 05:30:05,080 --> 05:30:06,520 EXCITED TO HAVE HER HERE TODAY. 7611 05:30:06,520 --> 05:30:08,640 HER TALK IS ENTITLED IN VIVO 7612 05:30:08,640 --> 05:30:10,280 ASSESSMENTS OF ADME PROPERTIES 7613 05:30:10,280 --> 05:30:15,680 OF LEAD COMPOUNDS. XIN, THE 7614 05:30:15,680 --> 05:30:26,000 FLOOR IS YOURS. 7615 05:30:30,800 --> 05:30:37,280 >>CAN YOU HEAR ME? I THANK YOU 7616 05:30:37,280 --> 05:30:40,480 FOR MY TALK ESPECIALLY AFTER 7617 05:30:40,480 --> 05:30:44,760 EXCITING TALKS MARC. WHAT I WILL 7618 05:30:44,760 --> 05:30:49,480 TALK ABOUT TODAY IS IN VITRO 7619 05:30:49,480 --> 05:30:57,840 ASSESSMENT OF THE ADME IS THE 7620 05:30:57,840 --> 05:30:59,640 FOUR IMPORTANT PROCESS OF DRUG 7621 05:30:59,640 --> 05:31:07,000 INVOLVEMENT. I WOULD LAKE TO 7622 05:31:07,000 --> 05:31:08,240 START ACKNOWLEDGMENT SLIDES, 7623 05:31:08,240 --> 05:31:09,360 COMBINED AWARD FROM MANY 7624 05:31:09,360 --> 05:31:12,320 SCIENTISTS IN THIS FIELD AND 7625 05:31:12,320 --> 05:31:16,760 ALSO LIKE TO THANK DR. 7626 05:31:16,760 --> 05:31:18,560 (INAUDIBLE) PHARMACOKINETIC 7627 05:31:18,560 --> 05:31:25,960 CHAPTER IN THIS ASSAY GUIDANCE. 7628 05:31:25,960 --> 05:31:27,600 HERE IS THE OUTLINE OF MY TALK. 7629 05:31:27,600 --> 05:31:33,760 FIRST GO OVER BASIC ADME 7630 05:31:33,760 --> 05:31:35,560 CONCEPTS IN DRUG DISCOVERY AND 7631 05:31:35,560 --> 05:31:39,880 DEVELOPMENT. AND I WILL GO OVER 7632 05:31:39,880 --> 05:31:44,200 SOME IN VITRO ADME ASSAYS IN THE 7633 05:31:44,200 --> 05:31:46,200 DRUG SELECTION LEAD COMPOUND AND 7634 05:31:46,200 --> 05:31:48,960 HELP DESIGN NEW MOLECULES IN THE 7635 05:31:48,960 --> 05:31:54,240 DRUG LIKE MOLECULES. I WILL TALK 7636 05:31:54,240 --> 05:31:56,560 ABOUT IN SILICO ADME BECAUSE 7637 05:31:56,560 --> 05:31:59,160 AFTER YOU GET THE DATA IN VITRO 7638 05:31:59,160 --> 05:32:00,560 ASSAYS, THE MODEL, ONCE YOU HAVE 7639 05:32:00,560 --> 05:32:02,920 A MODEL YOU CAN USE THE MODEL TO 7640 05:32:02,920 --> 05:32:05,000 START TO PREDICT OR HELP WITH 7641 05:32:05,000 --> 05:32:07,200 DESIGN THAT MAKES THE MOLECULE 7642 05:32:07,200 --> 05:32:08,640 AND I WILL END MY TALK WITH A 7643 05:32:08,640 --> 05:32:11,840 TAKE HOME MESSAGE. SO AS WITH E 7644 05:32:11,840 --> 05:32:14,160 KNOW, THERE ARE HURDLES FROM 7645 05:32:14,160 --> 05:32:16,240 DRUG DISCOVERY TO A FINAL 7646 05:32:16,240 --> 05:32:19,880 PRODUCT. SO WHAT WE ARE DOING 7647 05:32:19,880 --> 05:32:25,120 HERE ADME IS ONE HURDLE HERE. 7648 05:32:25,120 --> 05:32:27,560 WHAT HAPPEN IS BEFORE I GO TO 7649 05:32:27,560 --> 05:32:28,840 ADME I WILL TALK PHARMACO -- I 7650 05:32:28,840 --> 05:32:30,720 DON'T KNOW HOW MANY PEOPLE KNOW 7651 05:32:30,720 --> 05:32:33,800 WHAT IS THE PHARMACOKINETICS. IT 7652 05:32:33,800 --> 05:32:35,680 IS SCIENCE DESCRIBES THE DRUG 7653 05:32:35,680 --> 05:32:39,200 MOVEMENT IN THE BODY. WE USE A 7654 05:32:39,200 --> 05:32:40,360 MATHEMATICAL EQUATION TO 7655 05:32:40,360 --> 05:32:43,200 DESCRIBE THIS PROCESS. FOR 7656 05:32:43,200 --> 05:32:47,640 EXAMPLE, FOR ORAL YOU SEE AFTER 7657 05:32:47,640 --> 05:32:50,000 ORAL DOSE THE DIVIDE BY UPTAKE 7658 05:32:50,000 --> 05:32:54,160 OF IV DOSE DOSE. YOU CAN GET 7659 05:32:54,160 --> 05:32:56,840 ESTIMATE WHAT PERCENTAGE DRUG 7660 05:32:56,840 --> 05:32:59,120 INTO THE BIOLOGY SO MATHEMATIC 7661 05:32:59,120 --> 05:33:02,760 DESCRIBE CONCENTRATION AND TIME 7662 05:33:02,760 --> 05:33:04,120 RELATIONSHIP. WHY PHARMACO 7663 05:33:04,120 --> 05:33:06,040 KINETICS IS IMPORTANT? BACK IN 7664 05:33:06,040 --> 05:33:09,800 THE '90s YOU CAN SEE MAJORITY 7665 05:33:09,800 --> 05:33:12,040 OF DRUG FAILED IN THE CLINICAL 7666 05:33:12,040 --> 05:33:13,840 TRIALS DUE TO PHARMACO KINETICS. 7667 05:33:13,840 --> 05:33:17,240 SO HERE LIES THE IMPORTANT 7668 05:33:17,240 --> 05:33:18,960 PHARMACOKINETICS ALL PHARMACY 7669 05:33:18,960 --> 05:33:20,440 DISCOVERED RESEARCH PUT OWL 7670 05:33:20,440 --> 05:33:22,240 EFFORT IN PHARMACOKINETIC TRYING 7671 05:33:22,240 --> 05:33:25,000 TO SELECT COMPOUND FROM VERY 7672 05:33:25,000 --> 05:33:27,000 EARLY ON SO YOU CAN SAVE TIME 7673 05:33:27,000 --> 05:33:32,200 AND MONEY. SO SEEMS EFFECTIVELY 7674 05:33:32,200 --> 05:33:35,360 SEE THIS HIGH INCREASE 7675 05:33:35,360 --> 05:33:37,040 DRAMATICALLY, AS WE KNOW 20 7676 05:33:37,040 --> 05:33:42,120 YEARS AGO DATA TODAY IS ANY DUE 7677 05:33:42,120 --> 05:33:45,920 (INAUDIBLE) VERY, VERY LOW. HOW 7678 05:33:45,920 --> 05:33:50,920 WITH DO IT? THIS IS A CURVE THAT 7679 05:33:50,920 --> 05:33:52,240 SHOW PROFILE AFTER 7680 05:33:52,240 --> 05:33:54,440 ADMINISTRATION IN THE BODY. YOU 7681 05:33:54,440 --> 05:33:56,680 WANT THE CURVE HAS THE BLUE LINE 7682 05:33:56,680 --> 05:34:02,680 HERE, THAT MEANS THEY GAVE ABOVE 7683 05:34:02,680 --> 05:34:05,320 THE (INAUDIBLE) LEVEL BUT NOT 7684 05:34:05,320 --> 05:34:06,560 GOING TO BE HIGH TALKS LAB WHAT 7685 05:34:06,560 --> 05:34:09,360 IS REALLY THE FACTOR DETERMINING 7686 05:34:09,360 --> 05:34:11,280 THIS OCCUR AFTER GIVE ORAL 7687 05:34:11,280 --> 05:34:12,480 ADMINISTRATION, WE DIVIDE TO THE 7688 05:34:12,480 --> 05:34:17,160 FULL PROCESS, WE TALK ABOUT 7689 05:34:17,160 --> 05:34:22,440 ADME, ABSORPTION, DISTRIBUTION, 7690 05:34:22,440 --> 05:34:23,800 METABOLISM AND SECRETION. WE 7691 05:34:23,800 --> 05:34:26,160 WILL TALK ABOUT FACTORS IMPACT 7692 05:34:26,160 --> 05:34:27,920 WHOLE PROCESS. WE DEVELOP IN 7693 05:34:27,920 --> 05:34:32,800 VITRO ASSAYS. WE CAN COMBINE 7694 05:34:32,800 --> 05:34:35,360 THESE REGENERATING WHICH WE WILL 7695 05:34:35,360 --> 05:34:36,600 CALL RESULT BASED MODELING AND 7696 05:34:36,600 --> 05:34:38,240 PUT BACK MT. BODY BECAUSE WE 7697 05:34:38,240 --> 05:34:42,640 HAVE ALL THESE ORGANS THERE, AND 7698 05:34:42,640 --> 05:34:44,160 BROOD FLOW THERE, REGENERATED IN 7699 05:34:44,160 --> 05:34:46,080 VITRO YOU CAN PLUG IN AND YOU 7700 05:34:46,080 --> 05:34:51,520 CAN START TO PREDICT. THIS TIME 7701 05:34:51,520 --> 05:34:53,240 WE WILL TALK SMALL MOLECULE, 7702 05:34:53,240 --> 05:34:56,680 SMALL MOLECULE IDEALLY YOU GIVE 7703 05:34:56,680 --> 05:34:59,040 ORAL ADMINISTRATION DUE TO 7704 05:34:59,040 --> 05:35:00,280 SCREENS. FOUR OUR 7705 05:35:00,280 --> 05:35:02,320 ADMINISTRATION, THE DRUG WENT TO 7706 05:35:02,320 --> 05:35:03,800 SMALL BIOMARKERS GO THROUGH 7707 05:35:03,800 --> 05:35:07,840 SEVERAL BARRIERS IN ORDER TO 7708 05:35:07,840 --> 05:35:09,000 REALLY GET TARGET CONCENTRATION 7709 05:35:09,000 --> 05:35:12,360 WHEN WE WANT IT. THE FIRST IS 7710 05:35:12,360 --> 05:35:19,400 THE STOMACH. FOR COMPOUNDS NOT 7711 05:35:19,400 --> 05:35:20,760 CHEMICALLY STABLE THEY WILL 7712 05:35:20,760 --> 05:35:28,000 DEGREE DATE IN THE STOMACH. SO 7713 05:35:28,000 --> 05:35:31,440 ALSO SOLUBILITY IS IMPORTANTMENT 7714 05:35:31,440 --> 05:35:33,080 TO BE ABSORBED TO BLOODSTREAM 7715 05:35:33,080 --> 05:35:34,200 HAS TO BE SOLID ROLE SO 7716 05:35:34,200 --> 05:35:35,800 RATIONALE IN THE GI TRACK DOES 7717 05:35:35,800 --> 05:35:37,160 NOT HAVE A CHANCE TO BE ABSORBED 7718 05:35:37,160 --> 05:35:40,120 SO WE HAD TO LOOK AT SOLUBILITY 7719 05:35:40,120 --> 05:35:43,080 AT DIFFERENT PH. OTHER FACTOR 7720 05:35:43,080 --> 05:35:47,440 INVOLVE SUCH LIKE FULL -- 7721 05:35:47,440 --> 05:35:49,720 CONDITIONS IN (INAUDIBLE) EFFECT 7722 05:35:49,720 --> 05:35:50,920 SEVERITY FROM DIFFERENT SEGMENT 7723 05:35:50,920 --> 05:36:01,440 OF GI TRACK. THE SIMILAR PILOT 7724 05:36:04,160 --> 05:36:06,440 JUNCTION BETWEEN CELLS GIVE INTO 7725 05:36:06,440 --> 05:36:07,440 YOUR BLOOD CIRCULATION. SO 7726 05:36:07,440 --> 05:36:10,200 THERE'S SEVERAL PROCESS INVOLVED 7727 05:36:10,200 --> 05:36:12,440 IN THE ABSORPTION PROCESS HERE. 7728 05:36:12,440 --> 05:36:14,480 THE MOST COMMON ONE IS PASSIVE 7729 05:36:14,480 --> 05:36:16,600 DIFFUSION. THIS CONCENTRATION 7730 05:36:16,600 --> 05:36:19,800 GRADIENT PROCESS. THERE ARE MANY 7731 05:36:19,800 --> 05:36:22,280 DRUGS THAT WILL BE CALLED ACTIVE 7732 05:36:22,280 --> 05:36:23,720 TRANSPORTER INVOLVED. THEN FOR 7733 05:36:23,720 --> 05:36:26,640 THE LARGE MOLECULES THEY HAVE 7734 05:36:26,640 --> 05:36:28,880 MECHANISM CALLED ENDOCYTOSIS, 7735 05:36:28,880 --> 05:36:31,200 PIE NOCITOSIS, THE CELLS BROUGHT 7736 05:36:31,200 --> 05:36:33,880 INTO THE COMPOUND INTO THE NEXT 7737 05:36:33,880 --> 05:36:40,400 STEP OF ABSORPTION. ANOTHER 7738 05:36:40,400 --> 05:36:46,000 BARRIER IS ME TAB LIMB EFFLUX 7739 05:36:46,000 --> 05:36:47,800 TRANSPORTER STILL IN THE GI 7740 05:36:47,800 --> 05:36:52,080 TRACK HER HERE. WE KNOW THERE AE 7741 05:36:52,080 --> 05:36:54,920 MANY DRUG ENZYMES IN THE 7742 05:36:54,920 --> 05:36:56,360 ERYTHROSITES. HEPATOCYTES AND 7743 05:36:56,360 --> 05:36:58,960 THERE IS A TRANSPORTER THERE, 7744 05:36:58,960 --> 05:37:00,600 THEY CAN HAVE EVE FLUX. EVEN THE 7745 05:37:00,600 --> 05:37:04,000 DRUG INTO THE SYSTEM, THE 7746 05:37:04,000 --> 05:37:06,720 LUMINAL SIDE, BUT THEN THROUGH 7747 05:37:06,720 --> 05:37:09,400 THE EFFLUX TRANSPORTER CAN PUMP 7748 05:37:09,400 --> 05:37:15,160 UP SO STILL NOT INTO THE BLOOD 7749 05:37:15,160 --> 05:37:17,200 SIDE. NEXT ONE WILL BE THE 7750 05:37:17,200 --> 05:37:19,320 LIVER. LIVER IS LARGEST ORGAN, 7751 05:37:19,320 --> 05:37:21,480 HAVE ALL -- AFTER OF ALL THE 7752 05:37:21,480 --> 05:37:23,480 DRUG, METABOLISM ENZYME, THE 7753 05:37:23,480 --> 05:37:29,400 BIGGEST DEFENSE MECHANISM. 7754 05:37:29,400 --> 05:37:31,520 COMPOUND (INAUDIBLE) SO THE 7755 05:37:31,520 --> 05:37:34,720 LIVER HAS TWO ROWS THERE, ONE IS 7756 05:37:34,720 --> 05:37:36,560 THE ENZYME TRYING TO DO BILE 7757 05:37:36,560 --> 05:37:39,000 TRANSFORMATION, ANOTHER ONE IS 7758 05:37:39,000 --> 05:37:40,800 THROUGH THE SECRETION. THAT IS 7759 05:37:40,800 --> 05:37:42,400 REALLY THE IMPORTANT ORGAN WE 7760 05:37:42,400 --> 05:37:46,040 ALWAYS FOCUS ON WHEN WE LOOK AT 7761 05:37:46,040 --> 05:37:56,000 DRUG ABSORPTION AND (INAUDIBLE) 7762 05:37:56,000 --> 05:37:59,080 NOW YOU CAN SEE FOR DRUG TO BE 7763 05:37:59,080 --> 05:38:02,320 SUCCESSFULLY ABSORBED INTO THE 7764 05:38:02,320 --> 05:38:03,160 BLOOD CIRCULATION THEY HAVE THE 7765 05:38:03,160 --> 05:38:09,120 PASS SO MANY BARRIERS HER HER HE 7766 05:38:09,120 --> 05:38:12,320 BOWEL WILL BE PROCESS -- 7767 05:38:12,320 --> 05:38:13,920 BIOAVAILABILITY FOR THE ORAL 7768 05:38:13,920 --> 05:38:16,160 ABSORPTION, BAR IS PRETTY HIGH. 7769 05:38:16,160 --> 05:38:17,680 THE NEXT PARAMETER WE ARE 7770 05:38:17,680 --> 05:38:19,960 TALKING ABOUT IS TISSUE 7771 05:38:19,960 --> 05:38:21,640 DISTRIBUTION. MANY OF THESE 7772 05:38:21,640 --> 05:38:24,600 CHEMICAL PROPERTIES IMPACT THE 7773 05:38:24,600 --> 05:38:29,880 DISTRIBUTION, SUCH AS SIZE, 7774 05:38:29,880 --> 05:38:33,520 WEIGHT, CHARGE AND LIPO 7775 05:38:33,520 --> 05:38:36,720 PHYLLISTYTY. PROTEIN BINDING IS 7776 05:38:36,720 --> 05:38:38,880 ALSO A FACTOR. BLOOD IS DRIVER 7777 05:38:38,880 --> 05:38:40,560 DISTRIBUTE YOUR COMPOUND TO THE 7778 05:38:40,560 --> 05:38:44,600 TISSUES SO IF YOUR DRUG HAS HIGH 7779 05:38:44,600 --> 05:38:47,320 PROTEIN BINDING OR RESIDE INSIDE 7780 05:38:47,320 --> 05:38:49,880 RBC, AVAILABLE THE FREE DRUG 7781 05:38:49,880 --> 05:38:51,800 GAVE INTO THE TISSUE BENEFITS SO 7782 05:38:51,800 --> 05:38:57,160 WE'LL HAVE TO DETERMINE THAT. 7783 05:38:57,160 --> 05:38:59,160 ONCE DRUG IS INTO TISSUE SERVE 7784 05:38:59,160 --> 05:39:01,400 PURPOSE O TO DESIGN TARGETED 7785 05:39:01,400 --> 05:39:03,920 BINDING AS WE LIKE TO HAVE BUT 7786 05:39:03,920 --> 05:39:05,640 UNFORTUNATELY HAVE TISSUE 7787 05:39:05,640 --> 05:39:11,400 BINDING THERE. FOR SOME ONCE 7788 05:39:11,400 --> 05:39:15,360 BANDED TISSUE AND HARD FOR -- 7789 05:39:15,360 --> 05:39:22,960 THE NEW MODALITY OF (INAUDIBLE) 7790 05:39:22,960 --> 05:39:25,520 SO DRUG METABOLISM WE TALK ABOUT 7791 05:39:25,520 --> 05:39:27,840 IDEALLY SO SIMPLE BUT IF YOU 7792 05:39:27,840 --> 05:39:30,400 COME TO THE DETAILS SO MANY 7793 05:39:30,400 --> 05:39:33,600 REACTIONS GOING ON SO WE DIVIDE 7794 05:39:33,600 --> 05:39:35,480 TO TWO MAJOR CLASSES ONE CALLED 7795 05:39:35,480 --> 05:39:39,480 THE PHASE 1 METABOLISM, SECOND 7796 05:39:39,480 --> 05:39:43,560 IS PHASE 2 ME TAB LISP, PHASE 1 7797 05:39:43,560 --> 05:39:46,440 IS IF MOLECULE, THESE DAYS 7798 05:39:46,440 --> 05:39:48,360 MOLECULE TENDS TO BE POTENCY 7799 05:39:48,360 --> 05:39:52,280 DRIVEN. THAT MEANS VERY 7800 05:39:52,280 --> 05:39:53,600 PROPHETIC. HIGH AFFINITY FOR 7801 05:39:53,600 --> 05:40:00,160 TARGET. SO FOR THOSE MOLECULES 7802 05:40:00,160 --> 05:40:02,040 HARD TO PARTICIPANT SPLIT OUT. 7803 05:40:02,040 --> 05:40:04,840 POOR SOLUBILITY. THEY ARE TRYING 7804 05:40:04,840 --> 05:40:06,960 TO ADD SOME POLAR GROUP TO THE 7805 05:40:06,960 --> 05:40:09,600 MOLECULE THERE. SOFA SILL AT A 7806 05:40:09,600 --> 05:40:12,080 TIME EITHER GOING TOWARD PHASE 2 7807 05:40:12,080 --> 05:40:20,360 REACTION, THAT IS REALLY POLAR 7808 05:40:20,360 --> 05:40:23,280 GROWTH CELL BASED REALLY BECOME 7809 05:40:23,280 --> 05:40:24,640 SOLUBLE MOLECULE, THE MOLECULE 7810 05:40:24,640 --> 05:40:27,360 CAN BE EXCRETED TO THE BILE INTO 7811 05:40:27,360 --> 05:40:31,840 THE URINE. SO IT IS REALLY TWO 7812 05:40:31,840 --> 05:40:34,800 MAJOR FAMILIES OF DRUG 7813 05:40:34,800 --> 05:40:39,320 METABOLISM REACTIONS. FOR PHASE 7814 05:40:39,320 --> 05:40:41,880 1 METABOLISM, THERE AGAIN SO 7815 05:40:41,880 --> 05:40:44,240 MANY ENZYMES THERE IN 7816 05:40:44,240 --> 05:40:46,680 TRANSFORMATION REACTIONS THERE, 7817 05:40:46,680 --> 05:40:51,840 WHY WOULD ALWAYS -- MEDIATED 7818 05:40:51,840 --> 05:40:53,240 INTERACTION, BECAUSE LOOK AT THE 7819 05:40:53,240 --> 05:40:57,360 APPROPRIATE DRUG MORE THAN 75% 7820 05:40:57,360 --> 05:40:59,240 DRUG INVOLVE SIF METABOLISM SO 7821 05:40:59,240 --> 05:41:04,040 THEY BECOME VERY IMPORTANT. IN 7822 05:41:04,040 --> 05:41:06,760 ADDITION TO P 450 SOME 7823 05:41:06,760 --> 05:41:08,320 (INAUDIBLE) MEDIATED METABOLISM 7824 05:41:08,320 --> 05:41:11,800 ESPECIALLY WHEN WE CALL AD HIGH 7825 05:41:11,800 --> 05:41:14,680 OXIDASE. THEY SHARE SOME 7826 05:41:14,680 --> 05:41:18,520 SIMILARITY WHEN THEY DO IT 7827 05:41:18,520 --> 05:41:21,800 CHEMICAL REACTION AS P 450. 7828 05:41:21,800 --> 05:41:22,840 PEOPLE KNOW IT IS IMPORTANT 7829 05:41:22,840 --> 05:41:26,160 TRYING TO DESIGN THE MOLECULES 7830 05:41:26,160 --> 05:41:29,360 AVOID MEDIA METABOLISM SOMEHOW 7831 05:41:29,360 --> 05:41:31,400 PUSH SCAFFOLDS AND GOING TO BE 7832 05:41:31,400 --> 05:41:34,080 THE SUBSTRATE OF AO. SO WHEN WE 7833 05:41:34,080 --> 05:41:38,080 DO ALL THESE ASSAY DEVELOP WE 7834 05:41:38,080 --> 05:41:40,920 HAD TO KNOW WHICH FRACTION OF 7835 05:41:40,920 --> 05:41:45,960 THE CELL, ENZYMES. RESIDE IN THE 7836 05:41:45,960 --> 05:41:51,000 MECHANISM FRACTION, HOWEVER FOR 7837 05:41:51,000 --> 05:41:53,920 (INAUDIBLE) IN THE SIDE OF THE 7838 05:41:53,920 --> 05:41:56,880 FRACTION CELLS. FOR THE PEOPLE 7839 05:41:56,880 --> 05:42:04,560 NOW GOING TO P 450 IT IS FLAVIN 7840 05:42:04,560 --> 05:42:08,040 CONTAINING PROTEINS, THERE'S 7841 05:42:08,040 --> 05:42:10,280 SEVERAL HERE IF YOU HAVE NEW 7842 05:42:10,280 --> 05:42:12,000 DRUG FOR APPROVAL THEY PROBABLY 7843 05:42:12,000 --> 05:42:14,880 ASK YOU ALL THESE CYPs IN 7844 05:42:14,880 --> 05:42:17,240 TERMS OF ACTIVITY. BECAUSE THEY 7845 05:42:17,240 --> 05:42:22,880 WANT SO MANY DRUG SHARE ENZYME 7846 05:42:22,880 --> 05:42:24,480 HERE THEY WERE ABOUT TO HAVE ANY 7847 05:42:24,480 --> 05:42:31,040 DRUG INTERACTION SO THE COMMON 7848 05:42:31,040 --> 05:42:35,440 ONE IS 3, 4, THEY ACCOUNT FOR 7849 05:42:35,440 --> 05:42:38,160 ALMOST 50% OF THE DRUG THAT HAVE 7850 05:42:38,160 --> 05:42:43,280 THE MARKER INVOLVED. CYP 3A 4. 7851 05:42:43,280 --> 05:42:46,800 WE ALSO HAVE 3A 43A 5 ARE 7852 05:42:46,800 --> 05:42:49,880 SIMILAR, SHARE A LOT OF SAME 7853 05:42:49,880 --> 05:42:53,560 PROPERTY FOR DRUG METABOLISM. IN 7854 05:42:53,560 --> 05:42:55,680 ADDITION TO 4 AND 5, AND ALSO WE 7855 05:42:55,680 --> 05:42:58,960 ARE WORKING ON THE 3A 7, IT IS 7856 05:42:58,960 --> 05:43:00,920 THE ENZYME FOR INFANT AND AGE OF 7857 05:43:00,920 --> 05:43:06,880 ONE. SO FOR NEWBORN BABY, YOU 7858 05:43:06,880 --> 05:43:12,600 RARELY SEE THIS. AND ACTUAL ONE 7859 05:43:12,600 --> 05:43:15,840 YEAR OF AGE IS ACTIVITY DECREASE 7860 05:43:15,840 --> 05:43:19,280 IN 3A 4 START PICKING UP. WHEN 7861 05:43:19,280 --> 05:43:24,600 WE DO ALL THIS METABOLISM STUDY 7862 05:43:24,600 --> 05:43:25,480 REALIZE THE SPACIOUS DIFFERENCE 7863 05:43:25,480 --> 05:43:27,600 THERE BECAUSE WE START DOING PK 7864 05:43:27,600 --> 05:43:32,080 IN ANIMAL FIRST. THEN KNOW 7865 05:43:32,080 --> 05:43:33,720 ENZYMES THERE'S SPECIES 7866 05:43:33,720 --> 05:43:35,520 DIFFERENCE SO WE HAVEN'T GO 7867 05:43:35,520 --> 05:43:37,840 AHEAD AND MOVE FORWARD FOR THE 7868 05:43:37,840 --> 05:43:39,000 HUMAN THAT IS THE MECHANISM WE 7869 05:43:39,000 --> 05:43:43,800 ARE INTERESTED REALLY START 7870 05:43:43,800 --> 05:43:49,800 HUMAN ENZYME ASSAY EARLY ON. 7871 05:43:49,800 --> 05:43:51,360 FOR EXCRETION WE TALK ABOUT IT 7872 05:43:51,360 --> 05:43:54,040 BRIEFLY ALREADY, THE ADDITION 7873 05:43:54,040 --> 05:43:55,040 RANDOM EXCRETION IS ALSO 7874 05:43:55,040 --> 05:44:00,240 IMPORTANT. THE KEY FACTOR IMPACT 7875 05:44:00,240 --> 05:44:01,640 EXCRETION SIZE, AND 7876 05:44:01,640 --> 05:44:09,880 TRANSPORTERS. WE TALK ABOUT 7877 05:44:09,880 --> 05:44:13,800 MEDICINE IN NEED OF METABOLISM, 7878 05:44:13,800 --> 05:44:15,240 DURING COVID ONE CLASS COMPOUND 7879 05:44:15,240 --> 05:44:18,800 SHOW UP AS NEW ANALOG. OR 7880 05:44:18,800 --> 05:44:20,680 ANTIVIRAL DRUG. WE ALSO HAVE 7881 05:44:20,680 --> 05:44:22,520 METABOLISM FOR MANY DRUG, THAT 7882 05:44:22,520 --> 05:44:30,240 IS PRO DRUG. EXCRETION IS COPING 7883 05:44:30,240 --> 05:44:33,320 FOR. SO NOW TALK ABOUT FACTORS 7884 05:44:33,320 --> 05:44:34,800 THERE, THAT IS HOW GOING TO 7885 05:44:34,800 --> 05:44:38,720 ADDRESS FACTORS THAT DESIGN 7886 05:44:38,720 --> 05:44:44,120 BATTERY OF IN VITRO ASSAY. FOR 7887 05:44:44,120 --> 05:44:47,880 ABSORPTION, LOOK AT SOLUBILITY, 7888 05:44:47,880 --> 05:44:50,120 PERMEABILITY AND EFFLUX 7889 05:44:50,120 --> 05:44:51,800 TRANSPORTER IN INTESTINE. FOR 7890 05:44:51,800 --> 05:44:55,840 METABOLISM WE TALK ABOUT ALL 7891 05:44:55,840 --> 05:45:01,440 THESE P 450 METABOLISM OR OTHER 7892 05:45:01,440 --> 05:45:03,120 TRANSLOCATIONS THERE, AND ALSO 7893 05:45:03,120 --> 05:45:05,120 TALK ABOUT BILIARY SECRETION AND 7894 05:45:05,120 --> 05:45:08,000 RHYTHM SECRETION. SO ALL THESE 7895 05:45:08,000 --> 05:45:10,600 ASSAY, IN VITRO, GO IN THE 7896 05:45:10,600 --> 05:45:14,320 COMPOUND. FOR THE DISTRIBUTION 7897 05:45:14,320 --> 05:45:17,680 THOSE ARE VERY IMPORTANT. THAT 7898 05:45:17,680 --> 05:45:23,160 IS HOW FOCUS ON THESE IN VITRO 7899 05:45:23,160 --> 05:45:26,160 ASSAYS. SCREEN OUT MOLECULE. 7900 05:45:26,160 --> 05:45:27,520 ONLY THE WINNER ONE WILL BE 7901 05:45:27,520 --> 05:45:32,360 MOVED FORWARD TO THE NEXT STAGE 7902 05:45:32,360 --> 05:45:34,160 SO MANY ASSAYS IF YOU RUN FOR 7903 05:45:34,160 --> 05:45:35,720 EVERY COMPOUND, CALLS WILL BE 7904 05:45:35,720 --> 05:45:38,560 VERY HIGH SO THEY START 7905 05:45:38,560 --> 05:45:42,040 INVENTING WITH THE PARADIGM. I 7906 05:45:42,040 --> 05:45:43,160 BELIEVE MOST IN THIS WORKSHOP 7907 05:45:43,160 --> 05:45:44,600 PROBABLY DOING THIS CELL BASED 7908 05:45:44,600 --> 05:45:46,440 ASSAY OR SOME SAY ASSAYS FOR THE 7909 05:45:46,440 --> 05:45:50,080 TARGET. SO WHEN ADVOCACY 7910 05:45:50,080 --> 05:45:52,880 STARTED REALLY DOING SCREENING, 7911 05:45:52,880 --> 05:45:56,040 FOR DMPK FUNCTION WE ALSO START 7912 05:45:56,040 --> 05:45:59,360 SCREENING BUT THAT WE CALL -- WE 7913 05:45:59,360 --> 05:46:01,240 ARE LIMITED ONLY JUST A FEW 7914 05:46:01,240 --> 05:46:03,160 ASSAYS THERE. THAT IS THE BASIC 7915 05:46:03,160 --> 05:46:07,240 ONE. FOR EXAMPLE SOLUBILITY AND 7916 05:46:07,240 --> 05:46:08,560 PERMEABILITY AND MICROSOMAL 7917 05:46:08,560 --> 05:46:13,840 STABILITY. IN SOME CASES LOOK AT 7918 05:46:13,840 --> 05:46:17,320 SIP INHIBITION. SO ONLY WHEN 7919 05:46:17,320 --> 05:46:19,040 PAST THIS HIGH SUPER SCREENING 7920 05:46:19,040 --> 05:46:21,720 THERE, WE SEND TO THE TEAM AND 7921 05:46:21,720 --> 05:46:22,880 TEAM TAKE A LOT, THEY ARE GOING 7922 05:46:22,880 --> 05:46:26,360 TO DESIGN THE NEXT SIZE MOLECULE 7923 05:46:26,360 --> 05:46:30,280 WE CALL THE STRUCTURE 7924 05:46:30,280 --> 05:46:32,880 OPTIMIZATION. THOSE ACTIVITY 7925 05:46:32,880 --> 05:46:35,400 THERE. THEN YOU DIVE IN TO EACH 7926 05:46:35,400 --> 05:46:37,200 TEAM AND UNDERSTAND WHAT IS THE 7927 05:46:37,200 --> 05:46:39,040 TARGET, WHAT DRUG WE NEED TO GET 7928 05:46:39,040 --> 05:46:42,880 TO THAT TARGET, DESIGN TIER 2 7929 05:46:42,880 --> 05:46:46,680 ASSAY. THAT IS THE PROJECT ASSAY 7930 05:46:46,680 --> 05:46:49,800 THERE IS. BUT AFTER THAT WE LEAD 7931 05:46:49,800 --> 05:46:53,000 AND GO INTO THE IN VIVO. IN VIVO 7932 05:46:53,000 --> 05:46:55,000 USUALLY START WITH RODENT. 7933 05:46:55,000 --> 05:46:58,760 ESPECIALLYRODENT.ESPECIALLY YOUO 7934 05:46:58,760 --> 05:47:01,600 CONCEPT STARTING. WHATEVER IN 7935 05:47:01,600 --> 05:47:04,400 VITRO POTENCY LOOKING AT IN VIVO 7936 05:47:04,400 --> 05:47:08,320 ANIMAL DISEASE MODEL ELLE MODELK 7937 05:47:08,320 --> 05:47:10,360 THERE. ONLY AFTER YOU VALIDATE 7938 05:47:10,360 --> 05:47:11,360 TARGET THEN THE COMPANY, 7939 05:47:11,360 --> 05:47:12,480 MANAGEMENT BE MORE INTERESTED 7940 05:47:12,480 --> 05:47:15,240 REALLY PUT MORE MONEY THERE TO 7941 05:47:15,240 --> 05:47:17,640 SUSTAIN THAT PROGRAM. SO THE 7942 05:47:17,640 --> 05:47:19,800 PROOF CONCEPTS BECOME IMPORTANT. 7943 05:47:19,800 --> 05:47:23,280 THEN WE HAVE TO REALLY DETERMINE 7944 05:47:23,280 --> 05:47:24,520 PHARMACOKINETICS AT THAT STAGE 7945 05:47:24,520 --> 05:47:27,000 MAKE SURE THE DRUG WILL HAVE THE 7946 05:47:27,000 --> 05:47:29,120 CONCENTRATION ABOVE IN VITRO 7947 05:47:29,120 --> 05:47:30,720 ASSAY SPECIFIC ESPECIALLY IN THE 7948 05:47:30,720 --> 05:47:34,160 TARGET ORGAN. THEN COME THROUGH 7949 05:47:34,160 --> 05:47:37,400 REALLY THE NEXT STEP, THEY PASS 7950 05:47:37,400 --> 05:47:42,200 THAT AND DOING ENABLING STUDIES 7951 05:47:42,200 --> 05:47:49,400 AND REALLY (INAUDIBLE). WE WERE 7952 05:47:49,400 --> 05:47:50,960 FOCUSED ON THE ASSAY PART, 7953 05:47:50,960 --> 05:47:52,920 TALKING TIER 1 ASSAY, REALLY THE 7954 05:47:52,920 --> 05:47:54,640 HIGH THROUGH PUT SCREENING 7955 05:47:54,640 --> 05:47:57,640 ASSAYS. WHICH IS QUICKLY GO OVER 7956 05:47:57,640 --> 05:48:02,120 ALL COMPOUNDS SYNTHESIZED, OUT 7957 05:48:02,120 --> 05:48:03,320 IN TERMS OF KEY PARAMETERS 7958 05:48:03,320 --> 05:48:05,680 LISTED HERE, SOLUBILITY, 7959 05:48:05,680 --> 05:48:08,640 PERMEABILITY, MICROSOMAL 7960 05:48:08,640 --> 05:48:12,520 STABILITY AND CYPs SO IT IS 7961 05:48:12,520 --> 05:48:17,280 FASTER PACE, RAPID TURN AROUND 7962 05:48:17,280 --> 05:48:20,040 TIME SO COMBINATION IS SIMPLE. 7963 05:48:20,040 --> 05:48:25,600 NCATS WE USE A (INAUDIBLE) 7964 05:48:25,600 --> 05:48:26,880 PLATFORM AND SCREEN COMPOUNDS 7965 05:48:26,880 --> 05:48:29,480 SYNTHESIZED BY CHEMISTS, AVERAGE 7966 05:48:29,480 --> 05:48:32,240 100 COMPOUNDS PER WEEK. SO WE 7967 05:48:32,240 --> 05:48:33,520 SCREEN EVERY COMPOUND WITH THESE 7968 05:48:33,520 --> 05:48:36,560 THREE ASSAYS. THEN SOME COMPOUND 7969 05:48:36,560 --> 05:48:43,840 EVEN LOOK AT 3 OR 4. SO OVER THE 7970 05:48:43,840 --> 05:48:45,680 TEN YEARS NCATS JUST UNDER TEN 7971 05:48:45,680 --> 05:48:48,320 YEARS ANNIVERSARY SO FOR TIER 1 7972 05:48:48,320 --> 05:48:50,080 WE WERE TO HAVE DATA FOR OVER 7973 05:48:50,080 --> 05:48:56,400 30,000 COMPOUNDS. WE 7974 05:48:56,400 --> 05:48:58,720 (INAUDIBLE) BUT HOW WE ARE GOING 7975 05:48:58,720 --> 05:48:59,840 TO MANAGE HOW FOOD GOOD ASSAY 7976 05:48:59,840 --> 05:49:03,360 WILL BE, WE MONITOR EACH PLATE 7977 05:49:03,360 --> 05:49:06,760 AND THE VARIATION THERE, WE USE 7978 05:49:06,760 --> 05:49:09,280 THE MSR EARLY PRESENTATION IN 7979 05:49:09,280 --> 05:49:11,720 THIS WORKSHOP. SO THIS IS JUST 7980 05:49:11,720 --> 05:49:13,840 TABLE SUMMARIZE THE DATA WE HAVE 7981 05:49:13,840 --> 05:49:17,760 FOR OVER 500 PLACE. AND WHAT IS 7982 05:49:17,760 --> 05:49:19,680 THE MSR THERE. IN GENERAL THE 7983 05:49:19,680 --> 05:49:21,200 ASSAYS ARE PRETTY GOOD FOR TIER 7984 05:49:21,200 --> 05:49:27,480 1 ASSAY. ONCE YOU HAVE THE DATA 7985 05:49:27,480 --> 05:49:29,960 GO TO THE TEAM MEETING AND SEE 7986 05:49:29,960 --> 05:49:31,360 THESE TABLES VERY OFTEN, THAT 7987 05:49:31,360 --> 05:49:33,000 MEANS THE COMPOUNDS CHEMISTS 7988 05:49:33,000 --> 05:49:36,240 WITH THOSE COMPOUND STRUCTURE ON 7989 05:49:36,240 --> 05:49:37,880 YOUR (INAUDIBLE) THERE AND WE 7990 05:49:37,880 --> 05:49:40,440 HAVE ALL THESE AVME PROPERTIES 7991 05:49:40,440 --> 05:49:44,080 THERE AND THEY HAVE ALL THESE 7992 05:49:44,080 --> 05:49:46,560 ADVOCACY DATA THER THERE. SO THN 7993 05:49:46,560 --> 05:49:50,240 GAVE COLOR CODED, THE GREEN IS 7994 05:49:50,240 --> 05:49:53,480 THE ONE, THIS IS GOOD AND YELLOW 7995 05:49:53,480 --> 05:49:55,400 MEANS WARNING AND RED IS 7996 05:49:55,400 --> 05:49:56,640 DEFINITELY NOT GOING TO MOVE 7997 05:49:56,640 --> 05:50:00,640 FORWARD. THAT'S HOW BASED ON 7998 05:50:00,640 --> 05:50:02,040 THIS DATA THEY GO BACK TO SELECT 7999 05:50:02,040 --> 05:50:04,640 THE NEXT MOLECULE FOR NEXT 8000 05:50:04,640 --> 05:50:05,880 COMPOUND MOVING FORWARD. FOR 8001 05:50:05,880 --> 05:50:10,080 THE TIER 2 ASSAY AS MENTIONED 8002 05:50:10,080 --> 05:50:10,880 MORE SPECIFIC FOR THE PROJECT 8003 05:50:10,880 --> 05:50:12,440 NEEDS. IF YOU ARE DOING THE 8004 05:50:12,440 --> 05:50:15,440 PROJECT, THEN YOU PROBABLY GOING 8005 05:50:15,440 --> 05:50:24,360 TO LOOK AT STABILITY. LIKE 8006 05:50:24,360 --> 05:50:27,800 REMDESIVIR, SO MANY AS RACE 8007 05:50:27,800 --> 05:50:34,240 THERE. ANOTHER ASSAY WANT TO 8008 05:50:34,240 --> 05:50:36,040 MENTION HERE IS (INAUDIBLE) THAT 8009 05:50:36,040 --> 05:50:37,480 TELL YOU IF COMPOUND IS STABLE 8010 05:50:37,480 --> 05:50:40,440 WHICH MOLECULE IS PRONE FOR MOVE 8011 05:50:40,440 --> 05:50:43,480 METABOLISM, WE CALL THAT 8012 05:50:43,480 --> 05:50:47,360 METABOLIC (INAUDIBLE) ONCE YOU 8013 05:50:47,360 --> 05:50:50,640 IDENTIFY SOFT SPOT. TO GUIDE TO 8014 05:50:50,640 --> 05:50:55,600 BLOCK VULNERABLE SITE. THIS IS 8015 05:50:55,600 --> 05:51:00,600 ONE EXAMPLE WE USE TO SHOW HOW 8016 05:51:00,600 --> 05:51:03,320 THE MED IP GUIDE STRUCTURE OF 8017 05:51:03,320 --> 05:51:04,320 THE ORGANIZATION. IT IS ONE 8018 05:51:04,320 --> 05:51:08,240 PROJECT WE CALL FOP PROJECT 8019 05:51:08,240 --> 05:51:10,120 DISEASE, MET ID. OUR 8020 05:51:10,120 --> 05:51:11,600 COLLABORATOR ABOUT MASS GENERAL, 8021 05:51:11,600 --> 05:51:14,080 COME TO OUR, THEY THINK THEY HAD 8022 05:51:14,080 --> 05:51:16,680 LEAD COMPOUND ALREADY, THIS IS 8023 05:51:16,680 --> 05:51:20,440 STRUCTURE SHOWN HERE, LDN 8024 05:51:20,440 --> 05:51:24,720 193189. HOWEVER WHEN WE LOOK AT 8025 05:51:24,720 --> 05:51:26,440 IT, THAT HAS -- WE LOOK AT 8026 05:51:26,440 --> 05:51:28,120 STRUCTURE AND JUST DID SOME 8027 05:51:28,120 --> 05:51:30,320 SIMILAR, IT IS NOT COMPLETE 8028 05:51:30,320 --> 05:51:33,240 SCAFFOLD BUT ONE PORTION OF THE 8029 05:51:33,240 --> 05:51:34,400 MOLECULE WE LOOK AT THIS PART, 8030 05:51:34,400 --> 05:51:38,400 WE KNOW THERE IS SOME ADD 8031 05:51:38,400 --> 05:51:39,840 FORMATION THERE, WHAT WE CALL 8032 05:51:39,840 --> 05:51:41,440 DRAFT METABOLITE, THEY ARE NOT 8033 05:51:41,440 --> 05:51:43,040 STABLE, THEY ARE PROTEIN OR DNA, 8034 05:51:43,040 --> 05:51:44,920 WE CALL THAT REACTIVE 8035 05:51:44,920 --> 05:51:46,880 METABOLITES. SO WE SAY WELL WE 8036 05:51:46,880 --> 05:51:50,960 ARE NOT SURE YOUR MOLECULE WILL 8037 05:51:50,960 --> 05:51:53,720 BE -- BECAUSE WE ARE (INAUDIBLE) 8038 05:51:53,720 --> 05:51:55,360 AND WE SPEND SO MUCH TIME 8039 05:51:55,360 --> 05:51:56,640 CONVINCE PEOPLE THAT'S GOING TO 8040 05:51:56,640 --> 05:51:59,400 HAVE ISSUE, WE PUSH THE 8041 05:51:59,400 --> 05:52:01,680 (INAUDIBLE) STILL CANNOT PASS. 8042 05:52:01,680 --> 05:52:03,960 SO TRYING TO AVOID THIS KIND OF 8043 05:52:03,960 --> 05:52:05,640 THING HAPPENING SO WE SAY WHAT 8044 05:52:05,640 --> 05:52:07,960 DO WE DO WITH MET ID FIRST? THE 8045 05:52:07,960 --> 05:52:11,960 DATA COME BACK, WE SEE SEVERAL 8046 05:52:11,960 --> 05:52:15,400 LIABILITY FOR THIS MOLECULE. WE 8047 05:52:15,400 --> 05:52:19,480 SEE REACTIVE METABOLITE FORM. 8048 05:52:19,480 --> 05:52:23,400 ALSO SEE TRACKING AGENT. CAN 8049 05:52:23,400 --> 05:52:24,960 TELL YOU THERE'S REACTIVE 8050 05:52:24,960 --> 05:52:32,440 METABOLISM FORM. WE SEE FROM 8051 05:52:32,440 --> 05:52:38,400 ANOLI. THAT IS SPECIES RESEARCH, 8052 05:52:38,400 --> 05:52:40,440 ALWAYS SOME TOXICITY THERE. BUT 8053 05:52:40,440 --> 05:52:43,080 WHAT SURPRISED US MOST MAJOR 8054 05:52:43,080 --> 05:52:44,800 METABOLIC PATHWAY IS ON THIS 8055 05:52:44,800 --> 05:52:46,600 SIDE, OXIDATION SIDE. THIS 8056 05:52:46,600 --> 05:52:51,000 ENZYME IS REALLY NOT BY PEOPLE 8057 05:52:51,000 --> 05:52:54,680 50, RATHER BY AOO. WHY AO IS A 8058 05:52:54,680 --> 05:52:59,200 CONCERN THERE. THIS ENZYME, IT 8059 05:52:59,200 --> 05:53:01,920 IS SPECIES DEPENDENT. DOG 8060 05:53:01,920 --> 05:53:02,920 DOESN'T HAVE ACTIVITY FOR THIS 8061 05:53:02,920 --> 05:53:05,600 ENZYME BUT IF YOU LOOK AT 8062 05:53:05,600 --> 05:53:08,680 SPECIES SELECTION THESE DAYS YOU 8063 05:53:08,680 --> 05:53:11,440 HAVE MORE THAN (INAUDIBLE) BUT 8064 05:53:11,440 --> 05:53:13,760 THIS ONE IS NOT GOING TO WORK 8065 05:53:13,760 --> 05:53:15,680 BECAUSE THEY DON'T HAVE THIS 8066 05:53:15,680 --> 05:53:18,280 ENZYME ACTIVITY METABOLITE YOU 8067 05:53:18,280 --> 05:53:22,000 SEE IN HUMAN. EVEN IN THE 8068 05:53:22,000 --> 05:53:25,160 SPECIES LIKE RODENTS, AND HUMAN 8069 05:53:25,160 --> 05:53:27,600 HAVE ALL THESE ENZYME THERE, 8070 05:53:27,600 --> 05:53:29,080 THEY HAVE LARGE INTERSUBJECT 8071 05:53:29,080 --> 05:53:32,720 VARIATION, IN HUMANS AND SIX 8072 05:53:32,720 --> 05:53:33,640 MORE DIFFERENCE THERE. IMAGINE 8073 05:53:33,640 --> 05:53:35,440 SUCH A VARIATION THERE WILL BE 8074 05:53:35,440 --> 05:53:40,120 HARD FOR YOU GOING TO DECIDE 8075 05:53:40,120 --> 05:53:42,400 THOSE (INAUDIBLE) SO THERE IS A 8076 05:53:42,400 --> 05:53:44,720 LIABILITY THERE. AFTER THAT, THE 8077 05:53:44,720 --> 05:53:46,800 TEAM DECIDE WED GO BACK, WE ARE 8078 05:53:46,800 --> 05:53:49,240 GOING TO FIX ALL THESE 8079 05:53:49,240 --> 05:53:52,080 LIABILITIES. AFTER FEW YEARS 8080 05:53:52,080 --> 05:53:55,040 EFFORT WE DIDN'T COMPLETELY 8081 05:53:55,040 --> 05:53:58,040 ELIMINATE AO BUT ATTENUATED SO 8082 05:53:58,040 --> 05:54:02,520 THE MAJOR METABOLIC PATHWAY 8083 05:54:02,520 --> 05:54:05,080 CANONICAL IS (INAUDIBLE) THIS 8084 05:54:05,080 --> 05:54:07,000 COMPOUND IS IN CLINICAL PHASE 2 8085 05:54:07,000 --> 05:54:08,720 TRIAL IN EUROPE. SO THIS IS JUST 8086 05:54:08,720 --> 05:54:11,120 A STORY TELLING HOW MET ID CAN 8087 05:54:11,120 --> 05:54:15,480 BE NEWER STRUCTURE OPTIMIZATION. 8088 05:54:15,480 --> 05:54:17,480 THE OTHER STUDY IN THE TIER 2 8089 05:54:17,480 --> 05:54:21,360 ASSAY IS A CYP. THE CYP STUDY IS 8090 05:54:21,360 --> 05:54:24,280 COMPLICATED BECAUSE SO MANY 8091 05:54:24,280 --> 05:54:27,000 ISOFORMS THERE. FOR THE TRUE 8092 05:54:27,000 --> 05:54:29,960 LEAD COMES TO THE IND ENABLING 8093 05:54:29,960 --> 05:54:34,000 START YOU ARE ADDING SO YOU HAVE 8094 05:54:34,000 --> 05:54:35,880 TO FOLLOW ADME GUIDANCE FOR THE 8095 05:54:35,880 --> 05:54:38,280 DBI ASSESSMENT SO IN THAT THEY 8096 05:54:38,280 --> 05:54:40,120 LISTED THE ENZYME THEIR METADATA 8097 05:54:40,120 --> 05:54:43,440 SO YOU HAVE TO DO THE CYP 8098 05:54:43,440 --> 05:54:44,320 INHIBITION FOR THESE ENZYMES 8099 05:54:44,320 --> 05:54:48,600 THERE. I DON'T GO OVER DETAIL ON 8100 05:54:48,600 --> 05:54:51,000 THE ASSAY PROTOCOL NOW BECAUSE 8101 05:54:51,000 --> 05:54:56,840 YOU CAN ALL GET DETAILS ONLINE 8102 05:54:56,840 --> 05:54:59,120 BUT NEVERTHELESS CYP, IN THE 8103 05:54:59,120 --> 05:55:00,360 DISCOVERY STAGE HIGH THROUGH PUT 8104 05:55:00,360 --> 05:55:01,960 SCREENING WE PROBABLY JUST DO 8105 05:55:01,960 --> 05:55:06,000 ONE CONCENTRATION SAY 10 8106 05:55:06,000 --> 05:55:07,480 MICROMOLAR, PRETTY HIGH FOR IN 8107 05:55:07,480 --> 05:55:09,400 VIVO CONCENTRATION SO WE SAY AT 8108 05:55:09,400 --> 05:55:13,440 THAT CONCENTRATION IF YOU HAVE 8109 05:55:13,440 --> 05:55:15,120 IF YOU DON'T HAVE INHIBITION 8110 05:55:15,120 --> 05:55:16,560 THERE YOUR COMPOUND SAFE BUT IF 8111 05:55:16,560 --> 05:55:18,720 YOU HAVE THEN YOU MAY HAVE THE 8112 05:55:18,720 --> 05:55:21,960 TITRATE WITH CONCENTRATION TO 8113 05:55:21,960 --> 05:55:25,040 REALLY SEE HOW POTENT IT IS. SO 8114 05:55:25,040 --> 05:55:28,240 THAT IS THE DETAILED ASSAY 8115 05:55:28,240 --> 05:55:29,200 VERSUS HIGH THROUGH PUT 8116 05:55:29,200 --> 05:55:30,760 SCREENING ONE CONCENTRATION 8117 05:55:30,760 --> 05:55:32,840 MAYBE TEN MICROMOLAR, BEFORE THE 8118 05:55:32,840 --> 05:55:35,240 DETAILED ASSAY YOU HAVE TO HAVE 8119 05:55:35,240 --> 05:55:37,160 SEVERAL CONCENTRATIONS THERE, 8120 05:55:37,160 --> 05:55:39,000 BUT YOU CAN SIT CONCENTRATION 8121 05:55:39,000 --> 05:55:41,440 PUSH HIGH AS LIKE 50 OR 100 8122 05:55:41,440 --> 05:55:44,080 MICROMOLAR DEPENDS ON SOLUBILITY 8123 05:55:44,080 --> 05:55:45,920 BECAUSE SOME ANTIBIOTIC DRUGS, 8124 05:55:45,920 --> 05:55:52,120 THOSE ARE HIGH ESPECIALLY HIGH 8125 05:55:52,120 --> 05:55:57,360 POTENTIAL ON TDI TO EXTENT OF 8126 05:55:57,360 --> 05:55:59,160 OTHER DRUG SO WE HAVE TO HAVE 8127 05:55:59,160 --> 05:56:02,400 SAFETY WINDOW SO WE PUSH HIGH 8128 05:56:02,400 --> 05:56:04,600 CONCENTRATION AND BROAD RANGE 8129 05:56:04,600 --> 05:56:11,640 TIER 2 ASSAYS. ALSO IN THE 8130 05:56:11,640 --> 05:56:12,680 DEMONSTRATION YOU HAVE TO LOOK 8131 05:56:12,680 --> 05:56:15,400 AT SUICIDE INHIBITION. MOST 8132 05:56:15,400 --> 05:56:17,680 INHIBITION IS COMPETITIVE 8133 05:56:17,680 --> 05:56:19,640 INHIBITION, DRUG A DRUG B WORK 8134 05:56:19,640 --> 05:56:23,360 FOR THE SAME ENZYME, SOMETIMES 8135 05:56:23,360 --> 05:56:26,520 IF ONE DRUG A ENZYME 8136 05:56:26,520 --> 05:56:28,640 IRREVERSIBLE WE SAUL SUICIDE 8137 05:56:28,640 --> 05:56:32,360 ENZYME, SO WE NEED TO KNOW THAT 8138 05:56:32,360 --> 05:56:36,800 INFORMATION EARLY ON. THE NEXT 8139 05:56:36,800 --> 05:56:40,560 ASSAY IS THE INDUCTION. THAT 8140 05:56:40,560 --> 05:56:43,240 MEANS IF YOUR DRUG WERE INDUCED 8141 05:56:43,240 --> 05:56:45,600 CYP ACTIVITY IN VIVO, IS THERE 8142 05:56:45,600 --> 05:56:47,480 TWO CONCERNS THERE, ONE IS IF 8143 05:56:47,480 --> 05:56:51,280 YOU INDUCE YOUR ENZYME ACTIVITY, 8144 05:56:51,280 --> 05:56:54,400 YOUR DRUG (INAUDIBLE) STANDARD 8145 05:56:54,400 --> 05:56:58,520 DOSAGE THERE, NOW GAVE 8146 05:56:58,520 --> 05:56:59,840 EFFICACIOUS ANY MORE. THE SECOND 8147 05:56:59,840 --> 05:57:02,000 IS LONG TERM THIS INDUCTION MAY 8148 05:57:02,000 --> 05:57:05,360 HAVE A SAFETY CONCERN ESPECIALLY 8149 05:57:05,360 --> 05:57:08,400 ON THE LIVER AND THOSE TOXICITY 8150 05:57:08,400 --> 05:57:11,400 ISSUE THERE SO DETAILED GUIDANCE 8151 05:57:11,400 --> 05:57:14,760 THERE. FOR THE ASSAY, THERE'S 8152 05:57:14,760 --> 05:57:18,800 LOTS WE USE mRNA ASSAY OR DO 8153 05:57:18,800 --> 05:57:20,880 WE USE FUNCTIONAL ASSAY? THE 8154 05:57:20,880 --> 05:57:22,440 ARGUMENT IS THAT EVEN IF YOU 8155 05:57:22,440 --> 05:57:24,200 HAVE INDUCTION BUT IDEALLY HAVE 8156 05:57:24,200 --> 05:57:25,920 ACTIVITY THERE, SO IT IS NOT 8157 05:57:25,920 --> 05:57:28,680 GOING TO ALTER DOSE REGIM REGIMT 8158 05:57:28,680 --> 05:57:32,720 NOW OVER THE YEARS YOU CAN SEE 8159 05:57:32,720 --> 05:57:39,400 YOUR 2020 ADME GUIDANCE PRETTY 8160 05:57:39,400 --> 05:57:40,840 MUCH SETTLED IN mRNA ASSAY 8161 05:57:40,840 --> 05:57:42,760 THERE. THIS IS ASSAY GAVE YOU 8162 05:57:42,760 --> 05:57:45,320 HOW DO YOU CALCULATE, LOTS OF 8163 05:57:45,320 --> 05:57:46,920 DETAIL CALCULATIONS. SO FOR 8164 05:57:46,920 --> 05:57:49,880 PEOPLE REALLY COME TO IS THAT 8165 05:57:49,880 --> 05:57:52,600 STAGE ENCOURAGE GUIDANCE YOU 8166 05:57:52,600 --> 05:57:55,040 PROBABLY HAVE LEAD COMPOUND BONE 8167 05:57:55,040 --> 05:57:58,720 TO CANONICAL BUT LOTS OF DETAIL 8168 05:57:58,720 --> 05:58:04,760 IN THE DATA ON INDUCTION PART. 8169 05:58:04,760 --> 05:58:07,160 THESE ARE TWO GUIDANCE ISSUE IN 8170 05:58:07,160 --> 05:58:10,760 THE 2022. FIRST ONLY ONE 8171 05:58:10,760 --> 05:58:13,680 GUIDANCE ON -- BUT 2020 GUIDANCE 8172 05:58:13,680 --> 05:58:18,440 TWO, ONE IN VITRO FOR THE ASSAY 8173 05:58:18,440 --> 05:58:21,480 WORK YOU PROBABLY LOOK AT IN 8174 05:58:21,480 --> 05:58:24,800 VITRO PART BUT SECOND IS 8175 05:58:24,800 --> 05:58:28,760 CANONICACANONICAL. DRUG DRUG 8176 05:58:28,760 --> 05:58:31,640 INTERACTION ASSESSMENT. 8177 05:58:31,640 --> 05:58:34,320 TRANSPORTER ASSAY, OUR BODY HAS 8178 05:58:34,320 --> 05:58:39,120 SO MANY TRANSPORTERS, 8179 05:58:39,120 --> 05:58:41,960 (INAUDIBLE) INITIALLY WE ARE 8180 05:58:41,960 --> 05:58:45,760 INTERESTED IN PGP, PG FOR US IS 8181 05:58:45,760 --> 05:58:47,200 IMPACT WITH ABSORPTION, THAT CAN 8182 05:58:47,200 --> 05:58:52,080 PUMP IT OUT. SO THE SECOND PART 8183 05:58:52,080 --> 05:58:54,360 IS THE BRAIN UPTAKE T NOT -- IF 8184 05:58:54,360 --> 05:58:57,240 YOUR TARGET INSIDE, YOU HAVE 8185 05:58:57,240 --> 05:58:59,960 SOME TRAVEL THERE BUT NOW WE 8186 05:58:59,960 --> 05:59:06,400 KNOW ACTUALLY REALLY WORKING ON 8187 05:59:06,400 --> 05:59:08,880 THIS ANIMAL WE REALIZE ALL THESE 8188 05:59:08,880 --> 05:59:11,800 TISSUE DISTRIBUTIONS ARE IMPACT 8189 05:59:11,800 --> 05:59:15,240 BY TRANSPORTERS. IF YOU WORK ON 8190 05:59:15,240 --> 05:59:17,160 ASSAY DEVELOPMENT THAT IS -- YOU 8191 05:59:17,160 --> 05:59:21,000 CAN SPEND MORE TIME THAN COLLECT 8192 05:59:21,000 --> 05:59:25,400 DATA INFORMATION SO THESE ARE 8193 05:59:25,400 --> 05:59:26,920 STANDARD SLIDE SHOW THREE MAJOR 8194 05:59:26,920 --> 05:59:32,440 ORGANS THERE, THAT IS INTESTINE, 8195 05:59:32,440 --> 05:59:35,320 LIVER AND KIDNEY, ALL THESE 8196 05:59:35,320 --> 05:59:37,160 TRANSPORTERS CAN BE INVOLVED IN 8197 05:59:37,160 --> 05:59:40,280 EACH ORGAN. THIS IS PRETTY 8198 05:59:40,280 --> 05:59:44,480 COMPLICATED ALREADY, SO ALLOWS 8199 05:59:44,480 --> 05:59:46,480 TRANSPORTERS REGULATE AND THE 8200 05:59:46,480 --> 05:59:48,360 GUIDANCE SO THEY ARE DEMANDING 8201 05:59:48,360 --> 05:59:49,360 YOU HAVE TO SUBMIT ALL THIS 8202 05:59:49,360 --> 05:59:54,120 DATA. SO THAT IS REALLY THE 8203 05:59:54,120 --> 05:59:56,880 TRANSPORTER ASSESSMENT. BUT FOR 8204 05:59:56,880 --> 06:00:00,040 OURS AS RESEARCH AND WE WANT TO 8205 06:00:00,040 --> 06:00:01,320 UNDERSTAND HOW THESE TRANSPORTER 8206 06:00:01,320 --> 06:00:04,960 REALLY WORK. FOR EXAMPLE, ONE 8207 06:00:04,960 --> 06:00:09,080 NCATS PROJECT IS CYCLE 8208 06:00:09,080 --> 06:00:12,720 (INAUDIBLE) IN VITRO NOT MUCH 8209 06:00:12,720 --> 06:00:16,920 METABOLISM THERE, (INAUDIBLE) 8210 06:00:16,920 --> 06:00:19,680 ANIMAL PK INTO THE CANONICAL WE 8211 06:00:19,680 --> 06:00:23,200 SEE THE PEAK. SO WE KNOW IT IS 8212 06:00:23,200 --> 06:00:24,480 NOT DUE TO VASCULARIZATION, 8213 06:00:24,480 --> 06:00:26,280 BECAUSE THERE IS MOME TAB LIMB, 8214 06:00:26,280 --> 06:00:29,080 SO LIKELY DUE TO TRANSPORTER AND 8215 06:00:29,080 --> 06:00:32,160 ALSO DISTRIBUTION TRANSPORTER 8216 06:00:32,160 --> 06:00:33,760 ALONG THE INTESTINES. AGAIN WE 8217 06:00:33,760 --> 06:00:36,440 DON'T HAVE DATA TO DEMONSTRATE 8218 06:00:36,440 --> 06:00:39,080 THAT BUT JUST BRING PEOPLE'S 8219 06:00:39,080 --> 06:00:42,400 ATTENTION WORK ON ASSAY WILL 8220 06:00:42,400 --> 06:00:43,800 REALLY HELP YOU CAN REALLY WORK 8221 06:00:43,800 --> 06:00:46,360 ON THE TRANSPORTERS THAT WE ARE 8222 06:00:46,360 --> 06:00:47,640 GOING TO START COLLECTING THE 8223 06:00:47,640 --> 06:00:50,440 DATA LIKE IN THE EARLY 'T 90s 8224 06:00:50,440 --> 06:00:52,800 WE STARTED ON THE CYP REGULATION 8225 06:00:52,800 --> 06:00:59,000 GUIDANCE AND ALL THE CYP, 8226 06:00:59,000 --> 06:01:03,240 TRANSPORTERS. THIS ONE SHOW YOU 8227 06:01:03,240 --> 06:01:06,600 THE TRANSPORTERS HAVE SO MANY 8228 06:01:06,600 --> 06:01:08,280 DIFFERENT (INAUDIBLE) SOME 8229 06:01:08,280 --> 06:01:13,880 UPTAKE, SOME EFFLUX, SOME HAVE 8230 06:01:13,880 --> 06:01:17,280 BOTH ACTIONS IN AND OUT FOR THE 8231 06:01:17,280 --> 06:01:19,640 EFFLUX TRANSPORTER WE USE THIS 8232 06:01:19,640 --> 06:01:26,040 TRANSWELL SET UP BY ALSO HAD 8233 06:01:26,040 --> 06:01:31,160 TRANSWELLS. TRANSWELLS YOU HAVE 8234 06:01:31,160 --> 06:01:33,240 FROM (INAUDIBLE) AND MEMORY 8235 06:01:33,240 --> 06:01:34,840 HERE, CELL MEMBRANE AND THEN YOU 8236 06:01:34,840 --> 06:01:37,480 HAVE B SIDE HERE SO LOAD DRUG 8237 06:01:37,480 --> 06:01:40,920 FROM A SIDE DETERMINE DRUG ON 8238 06:01:40,920 --> 06:01:42,480 BOTH SIDE AFTER CERTAIN 8239 06:01:42,480 --> 06:01:44,640 INNERVATION TIME OR LOAD DRUG ON 8240 06:01:44,640 --> 06:01:47,080 B SIDE THIS CHANNEL HERE AND 8241 06:01:47,080 --> 06:01:48,480 LOOK AT DRUG CONCENTRATION ON 8242 06:01:48,480 --> 06:01:50,280 BOTH SIDES, THEN YOU CALCULATE 8243 06:01:50,280 --> 06:01:54,920 WHAT IS CALLED THE EFFLUX SO BY 8244 06:01:54,920 --> 06:02:00,240 BOTH REACTIONS AND THEN LOOK AT 8245 06:02:00,240 --> 06:02:01,240 EFFLUX RASH YES THERE. YOU HAVE 8246 06:02:01,240 --> 06:02:03,280 BACKGROUND NOISE BUT IF YOU 8247 06:02:03,280 --> 06:02:06,200 RATIO GREATER THAN 203 FOR SURE 8248 06:02:06,200 --> 06:02:10,280 YOU HAVE EFFLUX TRANSPORTERS. 8249 06:02:10,280 --> 06:02:13,440 WHEN WE DO THIS TRANSWELL ASSAY 8250 06:02:13,440 --> 06:02:17,880 WE NEED THE ASSESS MEMBRANE -- 8251 06:02:17,880 --> 06:02:19,120 ONE PARAMETER TO MEASURE IS 8252 06:02:19,120 --> 06:02:24,680 CALLED TIER MEASUREMENT. BECAUSE 8253 06:02:24,680 --> 06:02:27,240 IF MEMBRANE NOW FORM TIGHT 8254 06:02:27,240 --> 06:02:30,880 JUNCTION THERE, YOUR THE 8255 06:02:30,880 --> 06:02:32,960 TRANSPORT THROUGH MEMBRANE THERE 8256 06:02:32,960 --> 06:02:35,720 ACTUALLY IS A PATIVE DIFFUSION 8257 06:02:35,720 --> 06:02:36,400 BECAUSE YOU HAVE HOLE THERE. SO 8258 06:02:36,400 --> 06:02:40,360 YOU HAVE THE MAKE SURE YOU HAVE 8259 06:02:40,360 --> 06:02:41,760 THE BETWEEN CELL THAT FORM TIGHT 8260 06:02:41,760 --> 06:02:45,200 JUNCTION AND REALLY GOES LIEU 8261 06:02:45,200 --> 06:02:48,440 CELL MEMBRANE THROUGH DIFFUSION, 8262 06:02:48,440 --> 06:02:49,760 OR THROUGH TRANSPORTERS THERE. 8263 06:02:49,760 --> 06:02:54,880 TO MEASURE THE TIERS, MEASU 8264 06:02:54,880 --> 06:02:59,760 MEASUREMENT, BEFORE ONLY CELL 24 8265 06:02:59,760 --> 06:03:03,280 THE CELL TRANSPORT CELLS NOW YOU 8266 06:03:03,280 --> 06:03:10,560 SEE DRIVE FOR PACE AND RATE. 8267 06:03:10,560 --> 06:03:11,560 PROVIDE 96 WELL TRANSWELL 8268 06:03:11,560 --> 06:03:16,840 ALREADY. HOWEVER FOR THE PEER 8269 06:03:16,840 --> 06:03:18,120 MEASUREMENT, PRETTY CLASSICAL 8270 06:03:18,120 --> 06:03:21,800 THEY USE THE MANUAL ONE. REALIZE 8271 06:03:21,800 --> 06:03:28,400 THE GAP HERE NCATS SBR FUNDING 8272 06:03:28,400 --> 06:03:29,880 WE CONTACT APPLIED BASICS THAT 8273 06:03:29,880 --> 06:03:33,760 IS A COMPANY THAT HAVE 24 CELL 8274 06:03:33,760 --> 06:03:37,520 MEASUREMENT DEVICE THERE. WE ASK 8275 06:03:37,520 --> 06:03:38,400 IF THEY CAN MODIFY WHAT THEY 8276 06:03:38,400 --> 06:03:40,200 HAVE THERE AND MAKE FOR 96 8277 06:03:40,200 --> 06:03:47,440 WELLS. SO I SEE (INAUDIBLE) 8278 06:03:47,440 --> 06:03:52,400 THERE, MY GROUP, HE IS FORCE 8279 06:03:52,400 --> 06:03:58,200 BEHIND DEVICES. REALLY WORK WITH 8280 06:03:58,200 --> 06:04:05,880 COMPANY, CASCADE VALIDATE THE 8281 06:04:05,880 --> 06:04:09,920 DEVICE IS PRETTY MUCH IN 8282 06:04:09,920 --> 06:04:12,960 DATABASE. IN THIS DEVICE MEASURE 8283 06:04:12,960 --> 06:04:15,120 EACH WELL ONE SECOND, 8284 06:04:15,120 --> 06:04:16,720 AUGUSTN'TED TO FINISH 8285 06:04:16,720 --> 06:04:17,960 MEASUREMENT WE THINK TWO MINUTES 8286 06:04:17,960 --> 06:04:21,000 AND ALSO THIS DEVICE IS SO 8287 06:04:21,000 --> 06:04:23,680 COMPLEX THEY CAN PUT INSIDE 8288 06:04:23,680 --> 06:04:25,120 INCUBATOR SO YOU HAVE LOTS OF 8289 06:04:25,120 --> 06:04:26,000 FLUCTUATION TEMPERATURE WHEN YOU 8290 06:04:26,000 --> 06:04:32,120 MEASURE HERE. NOW I WILL TALK 8291 06:04:32,120 --> 06:04:38,120 ABOUT COMPLICATIONAL (INAUDIBLE) 8292 06:04:38,120 --> 06:04:43,120 BASED TOOL TO DESIGN AND PREDICT 8293 06:04:43,120 --> 06:04:50,280 DRUG CONTENT STRIAT CONCENTRATIS 8294 06:04:50,280 --> 06:04:55,800 NOT CHEAP. NEVERTHELESS WHEN WE 8295 06:04:55,800 --> 06:04:57,040 HAVE COLLABORATION WITH IP 8296 06:04:57,040 --> 06:05:00,640 CONSORTIUM IT IS A 8297 06:05:00,640 --> 06:05:01,520 PHARMACEUTICAL SCIENTIFIC WHEN 8298 06:05:01,520 --> 06:05:03,240 THEY GET TOGETHER AND TALK ABOUT 8299 06:05:03,240 --> 06:05:06,480 SCIENCE PART. THEY ALSO MENTION 8300 06:05:06,480 --> 06:05:09,400 VERY HELPFUL TO PUBLIC IF WE 8301 06:05:09,400 --> 06:05:13,840 HAVE SOME AVAILABLE SOFTWARE. SO 8302 06:05:13,840 --> 06:05:16,640 WITH THE DATA WE HAVE OVER TEN 8303 06:05:16,640 --> 06:05:19,960 YEARS AS I MENTION TIER 1 WE 8304 06:05:19,960 --> 06:05:27,360 HAVE DATA OVER 30,000 COMPOUNDS, 8305 06:05:27,360 --> 06:05:29,640 AT THIS MOMENT WE HAVE MODEL FOR 8306 06:05:29,640 --> 06:05:31,760 PARAMETERS SO THE NEXT STEP IS 8307 06:05:31,760 --> 06:05:35,880 TO START A WEBSITE WHERE WE 8308 06:05:35,880 --> 06:05:38,960 START DISSEMINATE MODELS, THESE 8309 06:05:38,960 --> 06:05:41,640 RETROSPECTIVELY VALIDATE AND 8310 06:05:41,640 --> 06:05:46,600 SUCCESS MARKET DRUGS, AND ALSO 8311 06:05:46,600 --> 06:05:47,760 THIS MODEL YOU CAN JOIN YOUR 8312 06:05:47,760 --> 06:05:52,720 STRUCTURE WITH ENZYME MOLECULE 8313 06:05:52,720 --> 06:05:55,120 OR (INAUDIBLE) THERE. FOR THE 8314 06:05:55,120 --> 06:05:59,840 PREDICTION OUTPUT, GIVE YOU SOME 8315 06:05:59,840 --> 06:06:00,760 CONFIDENCE OF FORCE AND 8316 06:06:00,760 --> 06:06:04,200 IMPORTANTLY YOU WANT PEOPLE USE 8317 06:06:04,200 --> 06:06:05,880 -- WANT SOMETHING BIG GUY BEHIND 8318 06:06:05,880 --> 06:06:07,040 THE SCREEN WE ARE DOING THERE SO 8319 06:06:07,040 --> 06:06:10,120 YOU CAN DOWNLOAD IT AND WE WILL 8320 06:06:10,120 --> 06:06:12,960 SEE DOWNLOAD YOU CAN REALLY 8321 06:06:12,960 --> 06:06:18,920 DESIGN AND PREDICT FOR YOURSELF 8322 06:06:18,920 --> 06:06:21,120 PROTOCOL REMEMBER TIER 1 ASSAY 8323 06:06:21,120 --> 06:06:27,320 ALL THESE PROTOCOL WE POSTED AT 8324 06:06:27,320 --> 06:06:29,480 (INAUDIBLE) TOGETHER WITH 8325 06:06:29,480 --> 06:06:32,240 PARTIAL DATA SETS, WE CANNOT 8326 06:06:32,240 --> 06:06:33,160 REROLLS ALL YET BECAUSE ALL 8327 06:06:33,160 --> 06:06:35,080 THESE ARE ACTIVE ONGOING 8328 06:06:35,080 --> 06:06:37,360 PROJECT. WE HAVE SOMETIME HAVE 8329 06:06:37,360 --> 06:06:39,320 THIS CREDA AGREEMENT AND WE 8330 06:06:39,320 --> 06:06:41,320 CANNOT RELEASE ALL STRUCTURE 8331 06:06:41,320 --> 06:06:44,200 FROM COMPOUND YET. THIS IS A 8332 06:06:44,200 --> 06:06:48,160 SCREEN SHOT OF THE WEBSITE. I 8333 06:06:48,160 --> 06:06:49,680 GAVE YOU THE WINDOW, YOU CAN 8334 06:06:49,680 --> 06:06:51,120 JOIN YOUR STRUCTURE HERE OR 8335 06:06:51,120 --> 06:06:53,960 REALLY DO THE BATCH, SOME -- 8336 06:06:53,960 --> 06:06:59,240 LIKE COMPANIES THEY HAVE DESIGN 8337 06:06:59,240 --> 06:07:00,920 COMPOUND PRECONDUCT SO WE CAN 8338 06:07:00,920 --> 06:07:05,280 HAVE STABILITY MANAGE UP TO 8339 06:07:05,280 --> 06:07:09,000 (INAUDIBLE). SO YOU CAN DOWNLOAD 8340 06:07:09,000 --> 06:07:10,760 IT, YOU DON'T NEED TO WORRY 8341 06:07:10,760 --> 06:07:16,880 ABOUT THE PRIVACY ISSUES THERE. 8342 06:07:16,880 --> 06:07:22,560 THIS IS JUST WE ARE -- CURRENTLY 8343 06:07:22,560 --> 06:07:25,400 WE HAVE NANOMOLARS THERE BUT AS 8344 06:07:25,400 --> 06:07:26,480 WE GROW WE WILL ADD MORE MODEL 8345 06:07:26,480 --> 06:07:31,880 THERE ITHERE IS. YOU CAN SEE MM 8346 06:07:31,880 --> 06:07:33,800 STABILITY, (INAUDIBLE) DIFFERENT 8347 06:07:33,800 --> 06:07:40,160 PH AND SOLUBILITY AND CELL 8348 06:07:40,160 --> 06:07:42,640 MICROSTABILITY THESE ARE 8349 06:07:42,640 --> 06:07:51,320 PROTOCOLS FOR ASSAYS, WE UPLOAD. 8350 06:07:51,320 --> 06:07:54,520 SO THAT ONE MODEL WHAT WE TALK 8351 06:07:54,520 --> 06:07:56,640 ABOUT HELP CHEMISTS DESIGN THE 8352 06:07:56,640 --> 06:07:58,480 STRUCTURE AND ALSO THESE DAYS 8353 06:07:58,480 --> 06:08:00,720 CAN DESIGN SO MANY, HOW MANY 8354 06:08:00,720 --> 06:08:02,120 REALLY MAKE. 8355 06:08:02,120 --> 06:08:04,520 SO REALLY HELP WITH THE GRAND 8356 06:08:04,520 --> 06:08:06,040 COMPOUND, PROBABLY GAVE THE ONE 8357 06:08:06,040 --> 06:08:08,920 LOOKS LIKE MORE LIKE A 8358 06:08:08,920 --> 06:08:11,760 (INAUDIBLE) HIGH PRIORITY FOR 8359 06:08:11,760 --> 06:08:17,200 THOSE STRUCTURE, WE HOPE WE CAN 8360 06:08:17,200 --> 06:08:20,880 ACCELERATE DISCOVERY OF THE TIME 8361 06:08:20,880 --> 06:08:23,040 NEEDED. ALSO THEY CAN HELP TO 8362 06:08:23,040 --> 06:08:25,080 EXPAND DISCREPANCY WHEN YOU DO 8363 06:08:25,080 --> 06:08:27,480 THE ENZYME VERSUS CELL BASED 8364 06:08:27,480 --> 06:08:31,640 ASSAY, SOMETIMES PROBABLY DUE TO 8365 06:08:31,640 --> 06:08:34,840 SOLUBILITY. P YOU HAVE STRAFER 8366 06:08:34,840 --> 06:08:35,920 MOLECULE WE CAN PREDICT 8367 06:08:35,920 --> 06:08:41,080 SOLUBILITY THEN YOU ARE SEEING 8368 06:08:41,080 --> 06:08:44,960 HOPEFULLY IN VITRO DATA THERE. 8369 06:08:44,960 --> 06:08:48,000 ALSO I KNOW THESE DAYS 8370 06:08:48,000 --> 06:08:51,120 ESPECIALLY DURING THE COVID 8371 06:08:51,120 --> 06:08:53,600 REPURPOSES LIBRARY SCREEN BECOME 8372 06:08:53,600 --> 06:08:55,840 POPULAR. SO COMPANIES START 8373 06:08:55,840 --> 06:08:57,040 PURCHASE BIG LIBRARY COMPOUNDS 8374 06:08:57,040 --> 06:08:58,560 THERE. . 8375 06:08:58,560 --> 06:09:00,960 SO BEFORE YOU DO PURCHASING YOU 8376 06:09:00,960 --> 06:09:02,760 SEE THESE TOOLS HOW TO HELP YOU 8377 06:09:02,760 --> 06:09:04,680 TO SELECT COMPOUNDS. SO THEY ARE 8378 06:09:04,680 --> 06:09:07,200 NOT EFFICACIOUS THERE. WHAT WE 8379 06:09:07,200 --> 06:09:11,920 TALK ABOUT NOW IS REALLY JUST IN 8380 06:09:11,920 --> 06:09:15,360 VITRO, DYNAMIC OPEN IN VITRO IS 8381 06:09:15,360 --> 06:09:17,840 CLOSED. SO AGAIN WE NEED TO HAVE 8382 06:09:17,840 --> 06:09:19,480 IN VITRO IN VIVO CORRELATION 8383 06:09:19,480 --> 06:09:22,720 THERE. IN VIVO PHARMACO -- THESE 8384 06:09:22,720 --> 06:09:23,720 ARE JUST COMMON PARAMETER WE 8385 06:09:23,720 --> 06:09:27,840 LOOK AT IN EXPOSURES, LIKE 8386 06:09:27,840 --> 06:09:30,600 VIABILITY ABSORPTION, 8387 06:09:30,600 --> 06:09:31,840 DISTRIBUTION, AND CLEARANCE AND 8388 06:09:31,840 --> 06:09:36,480 HALF LIVES AND SO ON. SO ONCE WE 8389 06:09:36,480 --> 06:09:41,520 GET ALL THESE IN VITRO ASSAY 8390 06:09:41,520 --> 06:09:46,040 ANIMAL DATA THERE, WE CAN START 8391 06:09:46,040 --> 06:09:49,240 THESE CELLS BASED MODEL. IF YOU 8392 06:09:49,240 --> 06:09:52,560 LOOK AT BUILDING LOCKS FOR THIS 8393 06:09:52,560 --> 06:09:54,000 MODEL, THE FOREST PART IS 8394 06:09:54,000 --> 06:09:55,480 EASIER, YOU CAN SEARCH FROM THE 8395 06:09:55,480 --> 06:09:57,040 THE FIRST PART IS EASIER, YOU 8396 06:09:57,040 --> 06:09:58,280 CAN SEARCH -- RELATED PARAMETER 8397 06:09:58,280 --> 06:10:00,320 FOR YOUR ANIMAL, THE ORGAN 8398 06:10:00,320 --> 06:10:05,280 VOLUME, BLOOD FLOW , TISSUES AND 8399 06:10:05,280 --> 06:10:07,760 ENZYMES AND ORGANS. YOU CAN DO 8400 06:10:07,760 --> 06:10:10,400 LIST RESEARCH FOR YOUR OWN DATA 8401 06:10:10,400 --> 06:10:13,800 AND WHEN WE TALK ABOUT SOME DRUG 8402 06:10:13,800 --> 06:10:16,680 BIOLOGICAL PHYSICAL CHEMICAL 8403 06:10:16,680 --> 06:10:17,720 PROPERTIES TALK ABOUT ALREADY, 8404 06:10:17,720 --> 06:10:20,840 FOR EXAMPLE IF IT IS COMPOUND 8405 06:10:20,840 --> 06:10:23,360 MADE YOU ALREADY HAVE PHYSICAL 8406 06:10:23,360 --> 06:10:29,280 CHEMICAL PROPERTY, SOLUBILITY 8407 06:10:29,280 --> 06:10:33,920 WEIGHT, PERMEABILITY. 8408 06:10:33,920 --> 06:10:35,680 (INAUDIBLE) ANIMAL STARTED 8409 06:10:35,680 --> 06:10:37,960 PROTOCOLS THERE, ANIMAL DATA 8410 06:10:37,960 --> 06:10:39,400 ALREADY SO FROM ALL THIS YOU 8411 06:10:39,400 --> 06:10:40,600 PLUG IN ALL THIS INFORMATION 8412 06:10:40,600 --> 06:10:43,160 INTO HERE AND CHANGE NOW TO THE 8413 06:10:43,160 --> 06:10:45,080 HUMAN MODEL RATHER THAN ANIMAL. 8414 06:10:45,080 --> 06:10:48,520 THE FIRST THING IS TO ACTUALLY 8415 06:10:48,520 --> 06:10:49,640 ESTABLISH MODEL WITH ANIMAL 8416 06:10:49,640 --> 06:10:52,440 BECAUSE THAT ANIMAL YOU CAN HAVE 8417 06:10:52,440 --> 06:10:55,320 DATA. WE USE (INAUDIBLE) TO 8418 06:10:55,320 --> 06:10:56,840 CONFIRM THE MODEL SO AFTER THAT 8419 06:10:56,840 --> 06:11:00,320 YOU CAN HAVE THE CONFIDENCE THAT 8420 06:11:00,320 --> 06:11:02,640 YOU CAN PREDICT. SO FOR THIS 8421 06:11:02,640 --> 06:11:04,080 KIND OF MODEL HAVE LOTS OF 8422 06:11:04,080 --> 06:11:05,680 APPLICATION. WE ARE JUST 8423 06:11:05,680 --> 06:11:07,480 STARTING DOING OUR LAB NOW BUT 8424 06:11:07,480 --> 06:11:09,600 YOU CAN THINK ABOUT PEOPLE 8425 06:11:09,600 --> 06:11:12,200 PUSHING PKPD MODEL, IF YOU DON'T 8426 06:11:12,200 --> 06:11:16,800 HAVE PK MODEL FORGET PD -- SO DO 8427 06:11:16,800 --> 06:11:21,560 PK FIRST THEN ADD PB COMPONENTS 8428 06:11:21,560 --> 06:11:22,800 WHATEVER TARGET YOU HAVE THERE. 8429 06:11:22,800 --> 06:11:28,120 THAT IS PKPD MODEL SO THE 8430 06:11:28,120 --> 06:11:29,520 PATIENT PK MODELING WE CAN USE 8431 06:11:29,520 --> 06:11:34,800 THE FORECAST OF HUMAN PK BEFORE 8432 06:11:34,800 --> 06:11:35,680 PHASE 1 CLINICAL TRIALS. WE CAN 8433 06:11:35,680 --> 06:11:38,760 ASSESS THE POPULATION SPECIAL 8434 06:11:38,760 --> 06:11:41,240 PHARMACOKINETIC AND DIFFERENT 8435 06:11:41,240 --> 06:11:43,880 POPULATIONS LIKE CHILDREN OR 8436 06:11:43,880 --> 06:11:48,080 PEOPLE HAS RENAL DEFICIENCY 8437 06:11:48,080 --> 06:11:50,320 HEPATIC FAILURE. HOWEVER TO TALK 8438 06:11:50,320 --> 06:11:52,880 ABOUT DBI YOU HAVE DRUG DRUG 8439 06:11:52,880 --> 06:11:54,680 INTERACTION THERE BUT THAT IS IN 8440 06:11:54,680 --> 06:11:57,120 VITRO BUT THESE ARE REALLY IF 8441 06:11:57,120 --> 06:11:58,920 THEY HAVE SOME POTENTIAL DRUG 8442 06:11:58,920 --> 06:12:03,040 DRUG INTERACTION HOW WE CAN 8443 06:12:03,040 --> 06:12:04,960 JUDGE THOSE REACTIONS TO USE 8444 06:12:04,960 --> 06:12:07,960 THIS MODEL TO DO SOME EXCISE 8445 06:12:07,960 --> 06:12:09,600 THERE AND FINALLY WE TALK ABOUT 8446 06:12:09,600 --> 06:12:12,400 PK PD OR EVEN TALKS RELATIONSHIP 8447 06:12:12,400 --> 06:12:13,840 WITH ZERO CONCENTRATION. THIS 8448 06:12:13,840 --> 06:12:18,360 IS THE MODELS. THAT IS PRETTY 8449 06:12:18,360 --> 06:12:21,240 MUCH ALL I HAVE HERE. I HOPE 8450 06:12:21,240 --> 06:12:23,160 THROUGH THIS TALK IF YOU LEARN 8451 06:12:23,160 --> 06:12:26,280 THINGS HOW WE DO USE IN VITRO 8452 06:12:26,280 --> 06:12:28,040 ADME ASSAY TO SUPPORT DRUG 8453 06:12:28,040 --> 06:12:30,080 DISCOVERY AND DEVELOPMENT ALL 8454 06:12:30,080 --> 06:12:35,680 THE WAY TO THE FINAL MARKET. 8455 06:12:35,680 --> 06:12:38,360 REALLY SEPARATED ASSAY STAGE 8456 06:12:38,360 --> 06:12:40,480 WISE IN THE DRUG DISCOVERY STAGE 8457 06:12:40,480 --> 06:12:42,080 WE USE HIGH THROUGHPUT PUTT 8458 06:12:42,080 --> 06:12:43,200 SCREEN ASSAYS BECAUSE YOU HAVE 8459 06:12:43,200 --> 06:12:44,040 SO MANY COMPOUNDS THERE YOU ARE 8460 06:12:44,040 --> 06:12:46,080 TRYING TO FILTER OUT WHAT ARE 8461 06:12:46,080 --> 06:12:50,600 THE SCAFFOLD, WHAT ARE THE 8462 06:12:50,600 --> 06:12:53,040 LIMITED COME POUNDS THERE, THAN 8463 06:12:53,040 --> 06:12:55,000 YOU DIVE TO THE TIER 2 ASSAY, 8464 06:12:55,000 --> 06:12:58,960 THAT IS REALLY GOING TO FOR YOU 8465 06:12:58,960 --> 06:13:03,600 TO CHOOSE THE IND SELECTIVE 8466 06:13:03,600 --> 06:13:05,320 NEEDS THERE AND WE CAN STORE ALL 8467 06:13:05,320 --> 06:13:08,720 IN VITRO DATA THOSE INFORMATION 8468 06:13:08,720 --> 06:13:12,120 CAN HELP BUILD THE MODEL, THAT 8469 06:13:12,120 --> 06:13:13,920 CAN PREDICT THE GUIDES TO THE 8470 06:13:13,920 --> 06:13:17,200 CLINICAL TRIALS. SO I WILL ENDS 8471 06:13:17,200 --> 06:13:22,000 THIS TALK AGAIN PROBABLY TO MAKE 8472 06:13:22,000 --> 06:13:25,240 DATA VALID YOU HAVE TO HAVE 8473 06:13:25,240 --> 06:13:26,240 (INAUDIBLE). SO NEED TO HAVE 8474 06:13:26,240 --> 06:13:28,760 CONTROLS AND REPLICATE RUNS FOR 8475 06:13:28,760 --> 06:13:31,080 EACH ASSAY. THANK YOU. 8476 06:13:31,080 --> 06:13:38,080 [APPLAUSE] 8477 06:13:38,080 --> 06:13:39,320 >>THANK YOU SO MUCH FOR A 8478 06:13:39,320 --> 06:13:45,360 WONDERFUL TALK. QUESTIONS. 8479 06:13:45,360 --> 06:13:46,840 >>THANKS FOR AMAZING 8480 06:13:46,840 --> 06:13:49,200 PRESENTATION. THAT WAS 8481 06:13:49,200 --> 06:13:51,800 INSIGHTFUL. FOR THE IN SILICO 8482 06:13:51,800 --> 06:13:53,800 ADME DETERMINATIONS, IS THAT THE 8483 06:13:53,800 --> 06:13:56,840 OUTPUT OF THAT, IS THAT 8484 06:13:56,840 --> 06:13:58,200 SUFFICIENT FOR ANY DOWNSTREAM 8485 06:13:58,200 --> 06:13:59,320 REGULATORY CONSIDERATIONS? OR DO 8486 06:13:59,320 --> 06:14:05,080 YOU HAVE TO DO THEM AGAIN? 8487 06:14:05,080 --> 06:14:07,720 >>(INAUDIBLE) 8488 06:14:07,720 --> 06:14:08,280 >>SORRY? 8489 06:14:08,280 --> 06:14:11,640 >>THIS IS EARLY STAGE. FILE IND 8490 06:14:11,640 --> 06:14:12,040 THROUGH THE DATA. 8491 06:14:12,040 --> 06:14:13,720 >>YOU STILL NEED THE WET LAB. 8492 06:14:13,720 --> 06:14:18,760 >>AS I SAID, IT IS NOT -- I 8493 06:14:18,760 --> 06:14:20,280 MEAN (INAUDIBLE) AND FOR EXAMPLE 8494 06:14:20,280 --> 06:14:23,200 YOU PROBABLY HAVE ANIMAL PK 8495 06:14:23,200 --> 06:14:25,360 DATA. YOU HAVE PK DATA ANY WAY, 8496 06:14:25,360 --> 06:14:28,000 LOOK AT EXPOSURE THERE. RIGHT? 8497 06:14:28,000 --> 06:14:29,640 SO YOU HAVE INFORMATION THERE 8498 06:14:29,640 --> 06:14:31,880 ALREADY. SO REALLY TO HELP YOU 8499 06:14:31,880 --> 06:14:36,200 TO SAY FOR EXAMPLE, HOW MUCH IS 8500 06:14:36,200 --> 06:14:38,960 DUE TO THE METABOLISM. AND IF 8501 06:14:38,960 --> 06:14:41,760 YOU HAVE THE LIVER FAILURE AND 8502 06:14:41,760 --> 06:14:44,440 HOW MUCH IMPACT IT WILL HAVE SO 8503 06:14:44,440 --> 06:14:47,680 YOUR IN VITRO DATA HELPS. THEY 8504 06:14:47,680 --> 06:14:49,280 HAVE ALL THESE SOFTWARES THEY DO 8505 06:14:49,280 --> 06:14:52,920 SOME MODELING THERE TOO. THEY 8506 06:14:52,920 --> 06:14:55,440 WILL SEE THERE'S ANYTHING THE 8507 06:14:55,440 --> 06:14:59,520 TRIGGER CHANGE THE LABEL, OR 8508 06:14:59,520 --> 06:15:02,000 EVEN ON THE MARKET. (INAUDIBLE) 8509 06:15:02,000 --> 06:15:02,440 BLACK BOX. 8510 06:15:02,440 --> 06:15:04,680 >>ESSENTIALLY A GREAT TOOL TO 8511 06:15:04,680 --> 06:15:06,360 HELP YOU SELECT WHICH COMPOUNDS 8512 06:15:06,360 --> 06:15:07,080 UP TO SUBSEQUENTLY -- 8513 06:15:07,080 --> 06:15:08,040 >>YES. 8514 06:15:08,040 --> 06:15:10,640 >>BUT YOU STILL HAVE TO DO WET 8515 06:15:10,640 --> 06:15:10,800 LAB. 8516 06:15:10,800 --> 06:15:13,120 >>ACTUALLY I FELT MOST PEOPLE 8517 06:15:13,120 --> 06:15:14,920 RESPOND AID SAY DEVELOPMENT PART 8518 06:15:14,920 --> 06:15:16,520 PROBABLY EARLY STAGE, EARLY 8519 06:15:16,520 --> 06:15:18,960 STAGE ACTUALLY IS REALLY FOR YOU 8520 06:15:18,960 --> 06:15:22,400 TO DESIGN THE MOLECULE. I CAN 8521 06:15:22,400 --> 06:15:24,640 GIVE EXAMPLE, THERE IS NO 8522 06:15:24,640 --> 06:15:30,680 (INAUDIBLE) HERE. WE HAVE A 8523 06:15:30,680 --> 06:15:33,280 (INAUDIBLE) THEY ARE A SMALL 8524 06:15:33,280 --> 06:15:35,800 COMPANY. CRO THAT COMPOUND YOU 8525 06:15:35,800 --> 06:15:38,000 CAN JOIN. THOSE PEOPLE FORM 8526 06:15:38,000 --> 06:15:39,000 SMALL COMPANY THEY HAVE LOTS OF 8527 06:15:39,000 --> 06:15:41,680 EXPERIENCE IN THE PHARMA 8528 06:15:41,680 --> 06:15:46,760 ALREADY. (INAUDIBLE) THEY MADE 8529 06:15:46,760 --> 06:15:50,440 200 COMPOUND IN ONE OF THE 8530 06:15:50,440 --> 06:15:53,680 (INAUDIBLE) IN CHINA. THEY HAD 8531 06:15:53,680 --> 06:15:56,760 DATA FOR 75 COMPOUNDS. SO NOW 8532 06:15:56,760 --> 06:16:00,720 THEY START (INAUDIBLE) THEN 8533 06:16:00,720 --> 06:16:04,440 AGAIN THERE IS A MODEL. SO HOW 8534 06:16:04,440 --> 06:16:11,240 ABOUT YOU GIVE (INAUDIBLE) YOU 8535 06:16:11,240 --> 06:16:14,040 KNOW HOW IT IS RIGHT? SO FOR 8536 06:16:14,040 --> 06:16:15,320 STABILITY WE HAVE 95% 8537 06:16:15,320 --> 06:16:16,640 PREDICTION. AGAIN WE DON'T SEE 8538 06:16:16,640 --> 06:16:18,200 THE STRUCTURE. 8539 06:16:18,200 --> 06:16:18,640 >> 8540 06:16:18,640 --> 06:16:20,360 (OFF MIC) 8541 06:16:20,360 --> 06:16:23,680 >>YEAH IT IS ONLINE. SINCE WE 8542 06:16:23,680 --> 06:16:25,120 ALL -- I DID HAVE THE NEWER 8543 06:16:25,120 --> 06:16:29,440 SLIDES, WE LAUNCHED NCATS CALLED 8544 06:16:29,440 --> 06:16:33,400 ADME NCATS, THERE YOU CAN GO YOU 8545 06:16:33,400 --> 06:16:35,240 CAN TRY THINGS WE JUST LAUNCH A 8546 06:16:35,240 --> 06:16:38,080 YEAR AGO LAST YEAR, NOW WE HAVE 8547 06:16:38,080 --> 06:16:42,760 A -- OVER 2500 (INAUDIBLE) 8548 06:16:42,760 --> 06:16:44,440 AROUND 77 COUNTRIES IN ONE YEAR 8549 06:16:44,440 --> 06:16:47,560 AND AMONG THAT 500 RETURNING 8550 06:16:47,560 --> 06:16:49,400 (INAUDIBLE) SO PEOPLE ARE 8551 06:16:49,400 --> 06:16:51,080 TRYING. AGAIN OR CONTINUE ADDING 8552 06:16:51,080 --> 06:16:55,200 MORE. THE BEAUTY IS THAT IS AN 8553 06:16:55,200 --> 06:16:58,520 EFFORT THEY HAVE A LIBRARY, BUT 8554 06:16:58,520 --> 06:16:59,920 HERE WE ARE CONTINUING TO ADDING 8555 06:16:59,920 --> 06:17:02,960 NEW DATA. ACTUALLY FOR PHARMA 8556 06:17:02,960 --> 06:17:06,960 BECAUSE THEY HAVE BIGGER 8557 06:17:06,960 --> 06:17:08,200 (INAUDIBLE) THEIR MODEL 24 HOURS 8558 06:17:08,200 --> 06:17:09,520 UPDATE BECAUSE EVERY DAY NEW 8559 06:17:09,520 --> 06:17:13,840 DATA COME IN. ONCE YOU HAVE THIS 8560 06:17:13,840 --> 06:17:17,720 DESCRIPTOR SET ALL YOU NEED IS 8561 06:17:17,720 --> 06:17:18,880 (INAUDIBLE) TRAINING VERIFYING 8562 06:17:18,880 --> 06:17:20,680 ALL THESE. ONCE YOU HAVE THE 8563 06:17:20,680 --> 06:17:23,320 MODALITY, MODULE SET IT UP, VERY 8564 06:17:23,320 --> 06:17:25,920 -- I DON'T THINK WE HAVE SO MANY 8565 06:17:25,920 --> 06:17:28,160 COMPOUNDS SO WE ARE DOING MAYBE 8566 06:17:28,160 --> 06:17:33,880 MONTHLY. SO BUT STILL OUR -- THE 8567 06:17:33,880 --> 06:17:37,960 BEAUTY OUR DATA IS IT IS COVER 8568 06:17:37,960 --> 06:17:40,320 OVER 300 PROJECTS THE SCAFFOLDS 8569 06:17:40,320 --> 06:17:43,440 IS A VERY DIVERSE CHEMICAL SPACE 8570 06:17:43,440 --> 06:17:46,360 HERE. THOUGH THEY HAVE MILLIONS 8571 06:17:46,360 --> 06:17:49,120 COMPOUNDS THERE. YOU CAN LOOK AT 8572 06:17:49,120 --> 06:17:52,240 THIS REALLY WHAT (INAUDIBLE) 8573 06:17:52,240 --> 06:17:54,880 YOUR STAFF FOCUS ON THAT TARGET 8574 06:17:54,880 --> 06:17:57,240 RIGHT? SO YOU MAY HAVE MORE BUT 8575 06:17:57,240 --> 06:17:58,800 WE ARE NOT WORKING ON THAT 8576 06:17:58,800 --> 06:18:00,600 DISEASE ANY MORE, WE WILL SWITCH 8577 06:18:00,600 --> 06:18:03,240 OURS SO THERE IS ACTUALLY WE DO 8578 06:18:03,240 --> 06:18:07,080 THIS DATA MODELING ASSESSMENT WE 8579 06:18:07,080 --> 06:18:08,760 CALL (INAUDIBLE) THE RECENT DATA 8580 06:18:08,760 --> 06:18:15,360 IS MOST VALUABLE. 8581 06:18:15,360 --> 06:18:17,000 >>I HAVE A QUESTION ABOUT THE 8582 06:18:17,000 --> 06:18:18,600 ADME NCATS ALSO. THAT SOUNDS 8583 06:18:18,600 --> 06:18:19,920 LIKE A REALLY INTERESTING TOOL 8584 06:18:19,920 --> 06:18:21,160 AND WONDERING WHICH DATA GOES 8585 06:18:21,160 --> 06:18:24,680 INTO IT, IS IT JUST YOUR FIRST 8586 06:18:24,680 --> 06:18:26,280 PASS SCREENING WHICH I THINK IT 8587 06:18:26,280 --> 06:18:30,680 INCLUDES PAMPA BUT NOT CACO 2 8588 06:18:30,680 --> 06:18:35,440 RIGHT? (OVERLAPPING SPEAKERS) 8589 06:18:35,440 --> 06:18:39,240 >>PAMPA IS 2 PH THAT, IS 8590 06:18:39,240 --> 06:18:40,800 ORIGINAL IN THE LAB. BUT LATER 8591 06:18:40,800 --> 06:18:44,240 ON REALIZED FOR G ABSORPTION YOU 8592 06:18:44,240 --> 06:18:47,400 NEEDS TO HAVE LOWER PH SO WE DID 8593 06:18:47,400 --> 06:18:51,040 (INAUDIBLE) WE FIGURED, CAN DO 8594 06:18:51,040 --> 06:18:55,240 MORE PH LEVEL BUT BUILD 8595 06:18:55,240 --> 06:18:58,000 DATABASE. SO WE FIGURED THAT 8596 06:18:58,000 --> 06:18:59,840 WOULD BE (INAUDIBLE) FIVE IS 8597 06:18:59,840 --> 06:19:03,560 REASONABLE RIGHT? SO THEN WE 8598 06:19:03,560 --> 06:19:07,000 CORRELATE WITH IN VIVO 8599 06:19:07,000 --> 06:19:07,960 BIOAVAILABILITY AND TOGETHER YOU 8600 06:19:07,960 --> 06:19:10,360 HAVE A CORRECT ORDER METABOLISM. 8601 06:19:10,360 --> 06:19:12,880 IF YOU JUST USE THAT ONE 8602 06:19:12,880 --> 06:19:13,560 (INAUDIBLE) SOMETIMES NOT 8603 06:19:13,560 --> 06:19:17,760 ACCURATE. BECAUSE A (INAUDIBLE) 8604 06:19:17,760 --> 06:19:19,360 ADDING METABOLISM DATA THERE YOU 8605 06:19:19,360 --> 06:19:20,120 HAVE BETTER PREDICTION. 8606 06:19:20,120 --> 06:19:21,240 >>THAT IS INTERESTING BECAUSE 8607 06:19:21,240 --> 06:19:22,480 YOU DONE HAVE ANY TRANSPORTERS 8608 06:19:22,480 --> 06:19:25,160 IN THERE. 8609 06:19:25,160 --> 06:19:28,040 >>NOT YET. TRANSPORTERS, OKAY. 8610 06:19:28,040 --> 06:19:29,680 >>GIVEN THE CACO 2. 8611 06:19:29,680 --> 06:19:33,200 >>CACO 2 WE HAVE A FEW HUNDRED 8612 06:19:33,200 --> 06:19:39,280 COMPOUND NOW BUT USUALLY FOR NEW 8613 06:19:39,280 --> 06:19:42,200 MODEL MINIMALLY YOU NEED A FEW 8614 06:19:42,200 --> 06:19:43,240 THOUSAND COMPOUNDS. THE FIRST 8615 06:19:43,240 --> 06:19:48,640 MODEL IS I THINK IT (INAUDIBLE) 8616 06:19:48,640 --> 06:19:50,760 THAT WE HAVE ABOUT OF 07,000 8617 06:19:50,760 --> 06:19:52,880 COMPOUNDCOMPOUNDS. EVEN WITH TE 8618 06:19:52,880 --> 06:19:57,400 DON'T FEEL COMFORTABLE BUT NOW 8619 06:19:57,400 --> 06:20:04,240 WE HAVE OVER 31, 3200 SOLID 8620 06:20:04,240 --> 06:20:04,520 COMPOUNDS, 8621 06:20:04,520 --> 06:20:07,840 >>THANK YOU. 8622 06:20:07,840 --> 06:20:09,480 >>I DON'T SEE ANY MORE 8623 06:20:09,480 --> 06:20:11,440 QUESTIONS. THANK YOU SO MUCH, 8624 06:20:11,440 --> 06:20:12,280 XIN FOR YOUR WONDERFUL 8625 06:20:12,280 --> 06:20:16,480 PRESENTATION. 8626 06:20:16,480 --> 06:20:16,880 [APPLAUSE] 8627 06:20:16,880 --> 06:20:18,520 >>I HAVE ONE MORE SLIDE TO 8628 06:20:18,520 --> 06:20:28,880 SHARE. IF YOU BEAR WITH ME. 8629 06:20:28,880 --> 06:20:31,280 THANK YOU FOR BEING HERE, THANKS 8630 06:20:31,280 --> 06:20:35,640 FOR PRESENTERS FOR AMAZING 8631 06:20:35,640 --> 06:20:37,600 TALKS. Y'ALL FOR ATTENDING. 8632 06:20:37,600 --> 06:20:40,680 WOULD LIKE YOU TO FILL OUT THE 8633 06:20:40,680 --> 06:20:42,480 FEEDBACK SURVEY WE ARE GOING TO 8634 06:20:42,480 --> 06:20:45,480 SEND THROUGH EMAIL, CORRECT? 8635 06:20:45,480 --> 06:20:47,520 WOULD LOVE TO HEAR YOUR 8636 06:20:47,520 --> 06:20:48,960 FEEDBACK. PLEASE TELL US WHAT WE 8637 06:20:48,960 --> 06:20:51,720 DID GOOD, WHAT WE DIDN'T DO 8638 06:20:51,720 --> 06:20:53,760 WELL, WHAT DO WE WANT TO HEAR, 8639 06:20:53,760 --> 06:20:57,040 MORE FROM ASSAY MANUAL WHERE WE 8640 06:20:57,040 --> 06:20:58,560 CAN GO FUTURE TO LOVE TO HEAR 8641 06:20:58,560 --> 06:21:01,360 YOUR FEEDBACK. THE VIDEO OF 8642 06:21:01,360 --> 06:21:04,000 THIS WORKSHOP WILL BE AID MADE 8643 06:21:04,000 --> 06:21:05,880 AVAILABLE ON THE NIH VIDEOCAST 8644 06:21:05,880 --> 06:21:09,320 WEBSITE, WE WILL SHARE THAT WITH 8645 06:21:09,320 --> 06:21:13,520 YOU AS WELL, WE WILL ALSO SHARE 8646 06:21:13,520 --> 06:21:15,000 THE LINKS I TACKED ABOUT OR 8647 06:21:15,000 --> 06:21:16,640 RESOURCES, THE ASSAY GUIDANCE 8648 06:21:16,640 --> 06:21:19,560 MANUAL HAS, THE PRESENTATIONS OF 8649 06:21:19,560 --> 06:21:21,640 THE SPEAKERS, AT THEIR 8650 06:21:21,640 --> 06:21:22,920 DISCRETION OF COURSE IF WILLING 8651 06:21:22,920 --> 06:21:26,800 TO SHARE WITH Y'ALL. AND THAT IS 8652 06:21:26,800 --> 06:21:30,960 IT. THANK YOU SO MUCH FOR 8653 06:21:30,960 --> 06:21:33,760 ATTENDING. HOPEFULLY THIS WAS 8654 06:21:33,760 --> 06:21:36,200 HELPFUL. AND HAVE A WONDERFUL 8655 06:21:36,200 --> 06:21:37,440 NIGHT. I WILL ADJOURN THE 8656 06:21:37,440 --> 06:21:37,880 MEETING. THANK YOU. 8657 06:21:37,880 --> 06:21:48,080 [APPLAUSE]