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Cov(2)MS: An Automated and Quantitative Matrix-Independent Assay for Mass Spectrometric Measurement of SARS-CoV-2 Nucleocapsid Protein

[Image: see text] The pandemic readiness toolbox needs to be extended, targeting different biomolecules, using orthogonal experimental set-ups. Here, we build on our Cov-MS effort using LC–MS, adding SISCAPA technology to enrich proteotypic peptides of the SARS-CoV-2 nucleocapsid (N) protein from tr...

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Detalles Bibliográficos
Autores principales: Van Puyvelde, Bart, Van Uytfanghe, Katleen, Van Oudenhove, Laurence, Gabriels, Ralf, Van Royen, Tessa, Matthys, Arne, Razavi, Morteza, Yip, Richard, Pearson, Terry, Drouin, Nicolas, Claereboudt, Jan, Foley, Dominic, Wardle, Robert, Wyndham, Kevin, Hankemeier, Thomas, Jones, Donald, Saelens, Xavier, Martens, Geert, Stove, Christophe P., Deforce, Dieter, Martens, Lennart, Vissers, Johannes P.C., Anderson, N. Leigh, Dhaenens, Maarten
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9773173/
https://www.ncbi.nlm.nih.gov/pubmed/36490367
http://dx.doi.org/10.1021/acs.analchem.2c01610
Descripción
Sumario:[Image: see text] The pandemic readiness toolbox needs to be extended, targeting different biomolecules, using orthogonal experimental set-ups. Here, we build on our Cov-MS effort using LC–MS, adding SISCAPA technology to enrich proteotypic peptides of the SARS-CoV-2 nucleocapsid (N) protein from trypsin-digested patient samples. The Cov(2)MS assay is compatible with most matrices including nasopharyngeal swabs, saliva, and plasma and has increased sensitivity into the attomole range, a 1000-fold improvement compared to direct detection in a matrix. A strong positive correlation was observed with qPCR detection beyond a quantification cycle of 30–31, the level where no live virus can be cultured. The automatable sample preparation and reduced LC dependency allow analysis of up to 500 samples per day per instrument. Importantly, peptide enrichment allows detection of the N protein in pooled samples without sensitivity loss. Easily multiplexed, we detect variants and propose targets for Influenza A and B detection. Thus, the Cov(2)MS assay can be adapted to test for many different pathogens in pooled samples, providing longitudinal epidemiological monitoring of large numbers of pathogens within a population as an early warning system.