Development of a colorimetric assay for the detection of SARS-CoV-2 3CLpro activity.
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Authors
Garland, Gavin D
Harvey, Robert F
Mulroney, Thomas E
Monti, Mie
Fuller, Stewart
Haigh, Richard
Gerber, Pehuén Pereyra
Barer, Michael R
Publication Date
2022-04-29Journal Title
Biochem J
ISSN
0264-6021
Publisher
Portland Press Ltd.
Volume
479
Issue
8
Pages
901-920
Type
Article
This Version
VoR
Physical Medium
Print
Metadata
Show full item recordCitation
Garland, G. D., Harvey, R. F., Mulroney, T. E., Monti, M., Fuller, S., Haigh, R., Gerber, P. P., et al. (2022). Development of a colorimetric assay for the detection of SARS-CoV-2 3CLpro activity.. Biochem J, 479 (8), 901-920. https://doi.org/10.1042/BCJ20220105
Abstract
Diagnostic testing continues to be an integral component of the strategy to contain the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) global pandemic, the causative agent of Coronavirus Disease 2019 (COVID-19). The SARS-CoV-2 genome encodes the 3C-like protease (3CLpro) which is essential for coronavirus replication. This study adapts an in vitro colorimetric gold nanoparticle (AuNP) based protease assay to specifically detect the activity of SARS-CoV-2 3CLpro as a purified recombinant protein and as a cellular protein exogenously expressed in HEK293T human cells. We also demonstrate that the specific sensitivity of the assay for SARS-CoV-2 3CLpro can be improved by use of an optimised peptide substrate and through hybrid dimerisation with inactive 3CLpro mutant monomers. These findings highlight the potential for further development of the AuNP protease assay to detect SARS-CoV-2 3CLpro activity as a novel, accessible and cost-effective diagnostic test for SARS-CoV-2 infection at the point-of-care. Importantly, this versatile assay could also be easily adapted to detect specific protease activity associated with other viruses or diseases conditions.
Keywords
COVID 19, Coronavirus, assay development, Antiviral Agents, COVID-19, Colorimetry, Coronavirus 3C Proteases, Gold, HEK293 Cells, Humans, Metal Nanoparticles, Peptide Hydrolases, Protease Inhibitors, SARS-CoV-2
Sponsorship
Medical Research Council (MR/P008801/1)
Identifiers
External DOI: https://doi.org/10.1042/BCJ20220105
This record's URL: https://www.repository.cam.ac.uk/handle/1810/338362
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