1 research outputs found
Cov<sup>2</sup>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 patient samples. The Cov2MS 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 Cov2MS 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