Journal
SLAS DISCOVERY
Volume 26, Issue 7, Pages 851-854Publisher
ELSEVIER SCIENCE INC
DOI: 10.1177/24725552211008862
Keywords
high-throughput screening; high-content screening; hit triaging; false-positive exclusion; orthogonal assay; counter assay; cellular health
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Small-molecule screening is a powerful method for identifying modulators of biological targets, but hit compounds causing assay interference can lead to false positives. Computational and experimental approaches are essential for flagging and eliminating unwanted compounds to prioritize high-quality hits for further analysis.
Small-molecule screening is a powerful approach to identify modulators of either specific biological targets or cellular pathways with phenotypic endpoints. A myriad of assay technologies are available to assess the activity of enzymes, monitor protein-protein interactions, measure transcription factor activity in reporter assays, or detect cellular features and activities using high-content imaging. A common challenge during small-molecule screening is, however, the presence of hit compounds generating assay interference, thereby producing false-positive hits. Thus, efforts are needed to uncover such interferences to prioritize high-quality hits for further analysis. This process encompasses (1) computational approaches to flag undesirable compounds, and (2) the use of experimental approaches like counter, orthogonal, and cellular fitness screens to identify and eliminate artifacts. In this brief guide, we provide an overview for first-time users, highlighting experimental screening strategies to prioritize high-quality bioactive hits from high-throughput screening/high-content screening (HTS/HCS) campaigns.
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