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Journal of Biomolecular Screening
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High-Throughput Screening by Mass Spectrometry: Comparison with the Scintillation Proximity Assay with a Focused-File Screen of AKT1/PKB{alpha}

Andrea K. Quercia

Department of Research Technologies, Bayer Pharmaceuticals Corporation, West Haven, Connecticut

William A. Lamarr

BioTrove Inc., Woburn, Massachusetts

Jayhyuk Myung

Department of Research Technologies, Bayer Pharmaceuticals Corporation, West Haven, Connecticut

Can C. Özbal

BioTrove Inc., Woburn, Massachusetts

James A. Landro

Department of Research Technologies, Bayer Pharmaceuticals Corporation, West Haven, Connecticut

Kevin J. Lumb

Department of Research Technologies, Bayer Pharmaceuticals Corporation, West Haven, Connecticut, kevin.lumb.b{at}bayer.com

Mass spectrometry is an emerging format for label-free high-throughput screening. The main limitation of mass spectrometry is throughput, due to the requirement to purify samples prior to ionization. Here the authors compare an automated high-throughput mass spectrometry (HTMS) system (RapidFireTM) with the scintillation proximity assay (SPA). The cancer therapy target AKT1/PKB{alpha} was screened against a focused library of kinase inhibitors and IC50 values determined for all compounds that exhibit > 50% inhibition. A selection of additional compounds that exhibited ≤ 50% inhibition in the primary screen was chosen as controls to confirm inactives. The selection of compounds is expected to identify common actives, common inactives, false positives, and false negatives. Agreement is found between HTMS and SPA in terms of primary hit identification and hit confirmation. (Journal of Biomolecular Screening 2007:473-480)

Key Words: high-throughput screening • mass spectrometry • SPA • false positives • false negatives

This version was published on June 1, 2007

Journal of Biomolecular Screening, Vol. 12, No. 4, 473-480 (2007)
DOI: 10.1177/1087057107300647
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