FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)

FlipGFP assay characterizes the intracellular drug target engagement to M(pro) and PL(pro) and can be performed in the biosafety level 1/2 settings. Here, we provide the detailed protocol for the cell-based FlipGFP assay to identify and characterize SARS-CoV-2 M(pro) and PL(pro) inhibitors. We descr...

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Detalles Bibliográficos
Autores principales: Tan, Haozhou, Hu, Yanmei, Wang, Jun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10156985/
https://www.ncbi.nlm.nih.gov/pubmed/37329507
http://dx.doi.org/10.1016/j.xpro.2023.102323
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author Tan, Haozhou
Hu, Yanmei
Wang, Jun
author_facet Tan, Haozhou
Hu, Yanmei
Wang, Jun
author_sort Tan, Haozhou
collection PubMed
description FlipGFP assay characterizes the intracellular drug target engagement to M(pro) and PL(pro) and can be performed in the biosafety level 1/2 settings. Here, we provide the detailed protocol for the cell-based FlipGFP assay to identify and characterize SARS-CoV-2 M(pro) and PL(pro) inhibitors. We describe steps for cell passage and seeding, transfection, addition of compounds, and their incubation and timing. We then detail the quantification of the fluorescence signal of the assay For complete details on the use and execution of this protocol, please refer to Ma et al.(1)
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spelling pubmed-101569852023-05-04 FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro) Tan, Haozhou Hu, Yanmei Wang, Jun STAR Protoc Protocol FlipGFP assay characterizes the intracellular drug target engagement to M(pro) and PL(pro) and can be performed in the biosafety level 1/2 settings. Here, we provide the detailed protocol for the cell-based FlipGFP assay to identify and characterize SARS-CoV-2 M(pro) and PL(pro) inhibitors. We describe steps for cell passage and seeding, transfection, addition of compounds, and their incubation and timing. We then detail the quantification of the fluorescence signal of the assay For complete details on the use and execution of this protocol, please refer to Ma et al.(1) Elsevier 2023-05-04 /pmc/articles/PMC10156985/ /pubmed/37329507 http://dx.doi.org/10.1016/j.xpro.2023.102323 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Tan, Haozhou
Hu, Yanmei
Wang, Jun
FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title_full FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title_fullStr FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title_full_unstemmed FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title_short FlipGFP protease assay for evaluating in vitro inhibitory activity against SARS-CoV-2 M(pro) and PL(pro)
title_sort flipgfp protease assay for evaluating in vitro inhibitory activity against sars-cov-2 m(pro) and pl(pro)
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10156985/
https://www.ncbi.nlm.nih.gov/pubmed/37329507
http://dx.doi.org/10.1016/j.xpro.2023.102323
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AT huyanmei flipgfpproteaseassayforevaluatinginvitroinhibitoryactivityagainstsarscov2mproandplpro
AT wangjun flipgfpproteaseassayforevaluatinginvitroinhibitoryactivityagainstsarscov2mproandplpro

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