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
Descripción
Sumario: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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