Journal
JOURNAL OF BIOMOLECULAR SCREENING
Volume 12, Issue 4, Pages 473-480Publisher
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057107300647
Keywords
high-throughput screening; mass spectrometry; SPA; false positives; false negatives
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Mass spectrometry is an ernerging 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 highthroughput mass spectrometry (HTMS) system (RapidFire (TM)) 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.
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