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

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 pat...

Full description

Saved in:
Bibliographic Details
Published in:Analytical chemistry (Washington) 2022-12, Vol.94 (50), p.17379-17387
Main Authors: 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
Format: Article
Language:English
Subjects:
Clinical Laboratory Techniques - methods
COVID-19
COVID-19 Testing
Humans
Mass Spectrometry - methods
Peptides
SARS-CoV-2
Sensitivity and Specificity
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 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 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.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.2c01610