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Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns

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 fac...

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Detalles Bibliográficos
Autores principales: Rothenaigner, Ina, Hadian, Kamyar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8293735/
https://www.ncbi.nlm.nih.gov/pubmed/33882754
http://dx.doi.org/10.1177/24725552211008862
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author Rothenaigner, Ina
Hadian, Kamyar
author_facet Rothenaigner, Ina
Hadian, Kamyar
author_sort Rothenaigner, Ina
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description 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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spelling pubmed-82937352021-08-06 Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns Rothenaigner, Ina Hadian, Kamyar SLAS Discov Article 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. SAGE Publications 2021-04-21 2021-08 /pmc/articles/PMC8293735/ /pubmed/33882754 http://dx.doi.org/10.1177/24725552211008862 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Article
Rothenaigner, Ina
Hadian, Kamyar
Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title_full Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title_fullStr Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title_full_unstemmed Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title_short Brief Guide: Experimental Strategies for High-Quality Hit Selection from Small-Molecule Screening Campaigns
title_sort brief guide: experimental strategies for high-quality hit selection from small-molecule screening campaigns
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8293735/
https://www.ncbi.nlm.nih.gov/pubmed/33882754
http://dx.doi.org/10.1177/24725552211008862
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