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lüll Apparent activity in high-throughput screening: origins of compound-dependent assay interference Thorne N; Auld DS; Inglese JCurr Opin Chem Biol 2010[Jun]; 14 (3): 315-24Expansive compound collections made up of structurally heterogeneous chemicals, the activities of which are largely undefined, present challenging problems for high-throughput screening (HTS). Foremost is differentiating whether the activity for a given compound in an assay is directed against the targeted biology, or is the result of surreptitious compound activity involving the assay detection system. Such compound interference can be especially difficult to identify if it is reproducible and concentration-dependent - characteristics generally attributed to compounds with genuine activity. While reactive chemical groups on compounds were once thought to be the primary source of compound interference in assays used in HTS, recent work suggests that other factors, such as compound aggregation, may play a more significant role in many assay formats. Considerable progress has been made to profile representative compound libraries in an effort to identify chemical classes susceptible to producing compound interference, such as compounds commonly found to inhibit the reporter enzyme firefly luciferase. Such work has also led to the development of practices that have the potential to significantly reduce compound interference, for example, through the addition of non-ionic detergent to assay buffer to reduce aggregation-based inhibition.|Artifacts[MESH]|Drug Discovery/*methods[MESH]|Enzyme Inhibitors[MESH]|High-Throughput Screening Assays/*methods[MESH]|Research Design[MESH]|Spectrometry, Fluorescence[MESH] |