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The Laboratory Robotics
Interest Group Mid Atlantic Chapter |
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The Laboratory Robotics Interest Group Mid Atlantic ChapterApril 1999 MeetingCombinatorial Chemistry & High Throughput ScreeningDate:
Wednesday, April 21, 1999
Agenda: The Social Period will feature vendor exhibits, food and refreshments. Members interested in presenting a poster are encouraged to do so. Open career positions at your company may be announced or posted. There is no fee to attend the meeting. Bring a business card to drop in the registration fishbowl - it eases registration and qualifies you for the rosewood pen set drawing.
Presentation: A New Reaction Block as the Core of an
Integrated System for High Throughput Automated Organic Synthesis. A new reaction block has been developed that forms the core of an integrated system for automated high throughput organic synthesis. Unlike many reactors previously described, this new block is capable of either solid- or solution-phase parallel synthesis. In addition, it can also be used for the cleavage step from split and pool synthesis using commercially available MicroKan™ reactors. The block offers large enough reaction scale to be used for lead optimization, yet delivers products into standard 96-well format for efficient post synthesis processing. The reaction block is based on a novel multiple valve mechanism with a single functional moving part, leading to low cost and high reliability. Maximum efficiency of this new reactor system is obtained from consolidated post synthesis processing regardless of synthesis method. Post synthesis processing is based on product collection into custom microtubes in 96-well format. Purpose built workstations process microtubes for weighing and selection for screening. This talk will describe the evolution and design of this new reactor system, along with its place in an efficient overall automated synthesis operation.
Presentation: ALPHAScreen a
novel non-radioactive homogeneous assay technology for uHTS This presentation will describe a novel assay technology termed Amplified Luminescent Proximity Homogeneous Assay, and its applications in drug discovery uHTS. The assay relies on two proprietary latex Donor and Acceptor beads (~200 nm) and generates highly amplified long life fluorescent signal when the beads are brought into proximity upon a biological binding event. The Donor bead contains a photosensitizer which converts ambient oxygen to the excited singlet state upon excitation by a laser at 680 nm. The Acceptor bead contains a thioxene derivative that reacts with the singlet oxygen rapidly and undergoes a quantitative first order decay with chemiluminescence at 370 nm, which is then transferred to the coexisting fluorephores through typical energy transfer process to generate fluorescence signal at 520-620 nm. Because of its short life time in aqueous solution (~4 usec), singlet oxygen diffuses to a distance of no more than ~200 nm. We have developed 11 ALPHA assays including kinase, protease, helicase, protein-protein, protein-DNA, PCR, SNP detection, as well as cAMP cell-based assay. I'll present data for representative assays in 384/1536 format using 1 to 20 ul volume. This technology enables quick miniaturization for uHTS without having to increase assay concentrations.
Presentation: A Solution-Phase High-Throughput Positional
Scanning Approach
Presentation: COMPARATIVE STUDY ON THE
FEASIBILITY OF THE ENZYME ASSAYS AND CELLULAR REPORTER GENE ASSAYS IN YEAST IN THE 96-,
384- AND 1536-WELL PLATE FORMAT Today’s demands in the drug discovery process force pharmaceutical companies to screen an ever increasing number of compounds against an increasing amount of targets. Consequently, miniaturization of established assays has become an advantageous means in high-throughput-screening (HTS) to meet reagent and sample cost, turnaround and space requirements. One way to realize this is to move from the standard 96-well plate to higher density microplate formats. This talk will describe the adaptation of two fluorescence-based assay systems to the challenges of the 1536-well plate format from both the engineer’s and biochemist’s point of view. We optimized liquid handling parameters of the micropipetting device JOBIwellTM (Jenoptik-Bioinstruments GmbH, Germany) using fluorescein-isothiocyanate as fluorescence dye. On this basis pipetting routines were established for an enzyme assay (?galactosidase) and a transcription assay in yeast (human progesterone receptor, hPR) in the 384- and 1536-well format. Finally, the experimental results were compared to those obtained in the well-established 96-well format. In all three assay formats, bioconversion of fluorescein-di-?D-galactopyranoside occurred as a function of the ?galactosidase concentration (in vitro assay) and the reporter gene expression showed the expected dependence on the ligand’s dose and affinity (yeast transcription assay), respectively. We conclude that miniaturization using the higher density 384- and 1536-well plate formats is advantageous as the next evolutionary step in HTS. JOBIwellTM proves to be a powerful tool for a careful adaptation of the liquid handling procedures.
Presentation: Combinatorial synthesis using the IRORI
Accutag system During my talk I will discuss our experiences with the IRORI Accutag high-throughput synthesis system. I'll cover our use of the system, its' strengths, weaknesses, and how we plan to use the IRORI system in the future.
Presentation: Development
of Fluorescence Polarization and FRET Assays for Tyrosine and Serine/Threonine Kinases Homogeneous fluorescence approaches such as FRET and FP are very important technologies for developing effective HTS assays for discovering therapeutic leads of kinases. This presentation will compare FRET and FP technologies for both tyrosine and serine/threonine kinase assays in terms of sensitivity, reliability and cost of reagents. The presentation will focus on discovery of a high affinity anti-phosphoserine antibody and applications of this antibody in the development of FP assays for a number of serine/threonine kinases. The preliminary screening data will also be discussed.
Exhibitors
Exhibitors - Click here for Exhibitor Information
Directions (on line directions at http://marriotthotels.com/EWRHO/): Take Route 287 to Exit 39 (Route 10 West). Go through one stoplight and take 1st u-turn.
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