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  • Review Article
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High-throughput screening assays for the identification of chemical probes

Nature Chemical Biology volume 3, pages 466–479 (2007)Cite this article

Abstract

High-throughput screening (HTS) assays enable the testing of large numbers of chemical substances for activity in diverse areas of biology. The biological responses measured in HTS assays span isolated biochemical systems containing purified receptors or enzymes to signal transduction pathways and complex networks functioning in cellular environments. This Review addresses factors that need to be considered when implementing assays for HTS and is aimed particularly at investigators new to this field. We discuss assay design strategies, the major detection technologies and examples of HTS assays for common target classes, cellular pathways and simple cellular phenotypes. We conclude with special considerations for configuring sensitive, robust, informative and economically feasible HTS assays.

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Figure 1: Assay categories and methodologies.
Figure 2: Influence of assay design on the measured biological response.
Figure 3: Assay performance and sensitivity.
Figure 4: Compound effects on assay outputs.

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Acknowledgements

We thank D. Leja and J. Fekces for illustrations. The authors are supported by the Molecular Libraries Initiative of the NIH Roadmap for Medical Research and the Intramural Research Program of the National Human Genome Research Institute, NIH.

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  1. US National Institutes of Health Chemical Genomics Center, National Institutes of Health, 9800 Medical Center Drive, Bethesda, 20892-3370, Maryland, USA

    James Inglese, Ronald L Johnson, Anton Simeonov, Menghang Xia, Wei Zheng, Christopher P Austin & Douglas S Auld

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  1. James Inglese
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Correspondence to James Inglese.

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Inglese, J., Johnson, R., Simeonov, A. et al. High-throughput screening assays for the identification of chemical probes. Nat Chem Biol 3, 466–479 (2007). https://doi.org/10.1038/nchembio.2007.17

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