Development of a colorimetric assay for the detection of SARS-CoV-2 3CLpro activity.
COVID 19
Coronavirus
assay development
Journal
The Biochemical journal
ISSN: 1470-8728
Titre abrégé: Biochem J
Pays: England
ID NLM: 2984726R
Informations de publication
Date de publication:
29 04 2022
29 04 2022
Historique:
received:
02
03
2022
revised:
04
04
2022
accepted:
04
04
2022
pubmed:
6
4
2022
medline:
26
4
2022
entrez:
5
4
2022
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 35380004
pii: 231110
doi: 10.1042/BCJ20220105
pmc: PMC9162461
doi:
Substances chimiques
Antiviral Agents
0
Protease Inhibitors
0
Gold
7440-57-5
Peptide Hydrolases
EC 3.4.-
Coronavirus 3C Proteases
EC 3.4.22.28
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
901-920Subventions
Organisme : Medical Research Council
ID : MC_UU_00025/7
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/P008801/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MR/P008801/ 1
Pays : United Kingdom
Organisme : Department of Health
Pays : United Kingdom
Informations de copyright
© 2022 The Author(s).
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