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dc.contributor.authorGarland, Gavin D
dc.contributor.authorHarvey, Robert F
dc.contributor.authorMulroney, Thomas E
dc.contributor.authorMonti, Mie
dc.contributor.authorFuller, Stewart
dc.contributor.authorHaigh, Richard
dc.contributor.authorGerber, Pehuén Pereyra
dc.contributor.authorBarer, Michael R
dc.contributor.authorMatheson, Nicholas J
dc.contributor.authorWillis, Anne E
dc.date.accessioned2022-06-24T23:30:55Z
dc.date.available2022-06-24T23:30:55Z
dc.date.issued2022-04-29
dc.identifier.issn0264-6021
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/338362
dc.description.abstractDiagnostic 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.
dc.format.mediumPrint
dc.publisherPortland Press Ltd.
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectCOVID 19
dc.subjectCoronavirus
dc.subjectassay development
dc.subjectAntiviral Agents
dc.subjectCOVID-19
dc.subjectColorimetry
dc.subjectCoronavirus 3C Proteases
dc.subjectGold
dc.subjectHEK293 Cells
dc.subjectHumans
dc.subjectMetal Nanoparticles
dc.subjectPeptide Hydrolases
dc.subjectProtease Inhibitors
dc.subjectSARS-CoV-2
dc.titleDevelopment of a colorimetric assay for the detection of SARS-CoV-2 3CLpro activity.
dc.typeArticle
dc.publisher.departmentDepartment of Medicine
dc.publisher.departmentDepartment of Pharmacology
dc.date.updated2022-06-24T10:02:51Z
prism.endingPage920
prism.issueIdentifier8
prism.publicationDate2022
prism.publicationNameBiochem J
prism.startingPage901
prism.volume479
dc.identifier.doi10.17863/CAM.85771
dcterms.dateAccepted2022-04-04
rioxxterms.versionofrecord10.1042/BCJ20220105
rioxxterms.versionVoR
dc.contributor.orcidWillis, Anne E [0000-0002-1470-8531]
dc.identifier.eissn1470-8728
rioxxterms.typeJournal Article/Review
pubs.funder-project-idMedical Research Council (MR/P008801/1)
cam.issuedOnline2022-04-21
cam.depositDate2022-06-24
pubs.licence-identifierapollo-deposit-licence-2-1
pubs.licence-display-nameApollo Repository Deposit Licence Agreement


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International