omation mbers - Society for Laboratory Automation and Screening
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omation mbers - Society for Laboratory Automation and Screening

omation mbers - Society for Laboratory Automation and Screening omation mbers - Society for Laboratory Automation and Screening

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WP030 Kurt Lund ACESystems, Inc. 135 Sixth Street Del Mar, California 92014 klund@abac.com The HTS Compound-Thawing Bottle Neck (and How to Avoid it With a New Processor) In terms of laboratory HTS efficiency, the thawing requirement for stored compounds presents itself as a huge bottleneck…until now with the advent of the new Thawing Processor System (TPS) from ACESystems, Inc. In the present paper the thawing behavior of solvents frozen in microplates is investigated in two types of experiments: (1) conventional thawing on the bench, and (2) rapid thawing with the TPS. It is also discussed how the TPS allows wells to be thawed, individually without disturbing the other wells in the plate, thus preserving precious compounds. The clearest observation from the bench tests (1) is that solvents in the wells remain largely frozen for hours after plate-removal from the freezer. Thus, for a center well of a 96-well microplate, even 3 hours was not enough time to effect 100% thawing on the bench. The rapid thawing experiment (2) was conducted using computer feedback control with the TPS. For DMSO in a standard one-ml well, frozen to -5˚ C, this resulted in 100% thawing in only 31⁄2 minutes, without causing excessive temperatures to the solvent. Overall, it is concluded that there can be great enhancement to laboratory efficiency with the Thawing Processor System, as well as large savings in compound preservation by thawing only selected wells in a microplate, instead of the whole plate. WP031 Joseph Machamer Molecular Devices 1311 Orleans Drive Sunnyvale, California 94089-1136 joe_machamer@moldev.com 206 Co-Author(s) Greg Kazan, Molecular Devices Tom Onofrey, Millipore Corp. Recent Developments in High Throughput, Turn-Key Solubility, and Permeability Compound Ranking Assays When combined with data from primary screening assays, measurement of the physiochemical properties of new chemical entities can provide a basis for the ranking of compounds based on their potential to be developed into viable drug candidates. The strategy of ranking the developability of new chemical entities (NCE) based on characterization by high throughput characterization of compound solubility and permeability is increasingly being applied in the drug discovery industry. We have developed automatable, turn-key assays to measure solubility and permeability that use Millipore membrane technology and a Molecular Devices SpectraMax absorbance microplate reader. The benefits of this approach include good correlation with low throughput, “gold standard” assays, no methods development, automation friendliness, reduced need for instrument expertise, and software flagging of data validity.

WP032 Colin Masui Symyx Technologies Life Sciences 3100 Central Expressway Santa Clara, California 95051 cmasui@symyx.com High Throughput Process Optimization for the Fine Chemical and Pharmaceutical Industries Symyx Technologies is currently applying its expertise in high throughput screening to develop fully integrated workflows for process optimization that greatly reduces the time and material needed to find the optimum conditions for a given organic transformation. Automated robotics is utilized in order to prepare catalysts and substrates and for post reaction workup and analytical preparation. The ability to run under a diverse set of conditions with temperature ranges from -20 to 200˚C and pressures up to 1500psi are capable using proprietary reactor formats (96 well Hip and PPR ® -48 reactors). Automated in situ injection of catalysts and substrates under reaction pressure is capable in the Parallel Pressure Reactor (PPR ® -48) System. The entire systems are completed by incorporation of the Symyx Renaissance software suite which delivers a complete, secure database environment to design experiments, control robotics, capture, store, recover, and explore the data and information from every stage of the workflow. WP033 Timothy McCauley Pierce Biotechnology, Inc. R&D 3747 N. Meridian Road Rockford, Illinois 61101 tim.mccauley@piercenet.com 207 Co-Author(s) Mahesh Mathrubutham, Sherri Z. Millis, Aric G. Morgan, Michael L. Stanaitis IQ ® Technology: An Automated HTS Assay for Screening Kinase, Phosphatase, and Protease Targets IQ ® Technology, a patent-pending homogeneous, universal detection platform originally designed for high throughput screening of kinases and phosphatases, has now been applied to screening of proteases. Representative enzymes from each of the major classes of proteases have been assayed in the IQ format. Enzyme activity and compound inhibition data will be presented for such enzymes as Trypsin, MMP-3 and Caspase-3. The technology has been tested in 96- to 384- to 1536-well microplate formats and is universally suited for automated screening systems. IQ Technology is a direct, non-competitive assay format that does not require antibodies or radioactive reagents. Fluorophore-labeled peptides are used as enzyme substrates. Kinase or phosphatase activity is quantified by direct measurement of the phosphorylation state of the substrate. For protease activity, cleavage is quantified by utilizing a peptide substrate containing a phospho-residue distal to the fluorphore. Protease assays result in cleavage of the substrate, which liberates the fluorphore-labeled terminus from the phosphorylated terminus. Cleavage is measured by the change in fluorescence intensity that occurs when a proprietary iron-containing compound binds specifically to phosphoryl groups on peptides and quenches the fluorescence. Ki’s generated using this platform correlate with published values. IQ Technology provides a universal method that can be used with any peptide sequence and is insensitive to high concentrations of ATP and substrate. The IQ Technology has been validated against a large number of detergents, organics, and other reagents found in reaction mixtures and has been optimized for high throughput applications with representative Z´ values of 0.7. POSTER ABSTRACTS

WP032<br />

Colin Masui<br />

Symyx Technologies<br />

Life Sciences<br />

3100 Central Expressway<br />

Santa Clara, Cali<strong>for</strong>nia 95051<br />

cmasui@symyx.com<br />

High Throughput Process Optimization <strong>for</strong> the Fine Chemical <strong>and</strong> Pharmaceutical Industries<br />

Symyx Technologies is currently applying its expertise in high throughput screening to develop fully integrated<br />

workflows <strong>for</strong> process optimization that greatly reduces the time <strong>and</strong> material needed to find the optimum<br />

conditions <strong>for</strong> a given organic trans<strong>for</strong>mation. Automated robotics is utilized in order to prepare catalysts <strong>and</strong><br />

substrates <strong>and</strong> <strong>for</strong> post reaction workup <strong>and</strong> analytical preparation. The ability to run under a diverse set of<br />

conditions with temperature ranges from -20 to 200˚C <strong>and</strong> pressures up to 1500psi are capable using proprietary<br />

reactor <strong>for</strong>mats (96 well Hip <strong>and</strong> PPR ® -48 reactors). Automated in situ injection of catalysts <strong>and</strong> substrates under<br />

reaction pressure is capable in the Parallel Pressure Reactor (PPR ® -48) System. The entire systems are completed<br />

by incorporation of the Symyx Renaissance software suite which delivers a complete, secure database<br />

environment to design experiments, control robotics, capture, store, recover, <strong>and</strong> explore the data <strong>and</strong> in<strong>for</strong>mation<br />

from every stage of the workflow.<br />

WP033<br />

Timothy McCauley<br />

Pierce Biotechnology, Inc.<br />

R&D<br />

3747 N. Meridian Road<br />

Rock<strong>for</strong>d, Illinois 61101<br />

tim.mccauley@piercenet.com<br />

207<br />

Co-Author(s)<br />

Mahesh Mathrubutham, Sherri Z. Millis,<br />

Aric G. Morgan, Michael L. Stanaitis<br />

IQ ® Technology: An Automated HTS Assay <strong>for</strong> <strong>Screening</strong> Kinase, Phosphatase, <strong>and</strong><br />

Protease Targets<br />

IQ ® Technology, a patent-pending homogeneous, universal detection plat<strong>for</strong>m originally designed <strong>for</strong> high throughput<br />

screening of kinases <strong>and</strong> phosphatases, has now been applied to screening of proteases. Representative enzymes<br />

from each of the major classes of proteases have been assayed in the IQ <strong>for</strong>mat. Enzyme activity <strong>and</strong> compound<br />

inhibition data will be presented <strong>for</strong> such enzymes as Trypsin, MMP-3 <strong>and</strong> Caspase-3. The technology has been<br />

tested in 96- to 384- to 1536-well microplate <strong>for</strong>mats <strong>and</strong> is universally suited <strong>for</strong> automated screening systems.<br />

IQ Technology is a direct, non-competitive assay <strong>for</strong>mat that does not require antibodies or radioactive reagents.<br />

Fluorophore-labeled peptides are used as enzyme substrates. Kinase or phosphatase activity is quantified by direct<br />

measurement of the phosphorylation state of the substrate. For protease activity, cleavage is quantified by utilizing<br />

a peptide substrate containing a phospho-residue distal to the fluorphore. Protease assays result in cleavage of the<br />

substrate, which liberates the fluorphore-labeled terminus from the phosphorylated terminus. Cleavage is measured<br />

by the change in fluorescence intensity that occurs when a proprietary iron-containing compound binds specifically<br />

to phosphoryl groups on peptides <strong>and</strong> quenches the fluorescence. Ki’s generated using this plat<strong>for</strong>m correlate with<br />

published values. IQ Technology provides a universal method that can be used with any peptide sequence <strong>and</strong><br />

is insensitive to high concentrations of ATP <strong>and</strong> substrate. The IQ Technology has been validated against a large<br />

number of detergents, organics, <strong>and</strong> other reagents found in reaction mixtures <strong>and</strong> has been optimized <strong>for</strong> high<br />

throughput applications with representative Z´ values of 0.7.<br />

POSTER ABSTRACTS

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