Development and translation of a method of clinical utility for LC-MS/MS analysis to detect SARS-CoV-2 antigens from ONP swabs and saliva
Year: 2024
Type: preprint
Abstract: During the COVID-19 pandemic, development of diagnostic tests was vital to chart the course and to reduce the impact of the infection. Continued testing and surveillance of vaccine escape will continue for years to come which presents an opportunity to integrate such testing into clinical biochemistry laboratories that form part of integrated healthcare testing. Here we describe a protocol for a targeted mass spectrometry based proteomic assay (COVIDCAP) developed to detect SARS-CoV-2 peptides from oro-nasopharyngeal swabs (ONP) and saliva. This uses novel SISCAPA antibodies bound to magnetic beads and subsequent analysis of captured and purified SARS-CoV-2 nucleocapsid (NCAP) peptides. The method involves immediate deactivation of the sample using an ethanolic solution. This simultaneously inactivates the virus and denatures viral proteins at sampling in contrast to the approach for RT-PCR testing, with benefits for the assay as well as for downstream processing. A plate-based preparation of the samples involving acetone precipitation followed by a short tryptic digestion and subsequent immunocapture allows LC-MS detection and quantification of peptides from the NCAP protein, in a 3-minute inject-to-inject assay with an LOD of 20 attomoles from starting sample. For 576 ONP swab samples taken as exemplars here, the sensitivity and specificity of this analysis is shown to be 97.0% and 96.6% respectively.
Source: ChemRxiv
Authors: N Leigh Anderson, Alistair Bailey, Perdita Barran, Philip Brownbridge, Kathleen Cain, Rachel S Carling, Rainer Cramer, R Neil Dalton, Matthew E Daly, Kayleigh Davis, Ivan Doykov, Maria Emilia Dueñas, Edward Emmott, Claire E Eyers, Akshada Gajbhiye, Bethany Geary, Pankaj Gupta, Jenny Hällqvist, Evita Hartmane, Simon Heales, Tiaan Heunis, Wendy Heywood, Katherine A Hollywood, Rosalind E Jenkins, Donald JL Jones, Brian Keevil, Henriette Krenkel, Dan Lane, Catherine S Lane, Sophie Lellman, Ellen N Liggett, Xiaoli Meng, Clare Mills, Kevin Mills, Atakan A Nalbant, Leong L Ng, Benjamin Nicholas, Dan Noels, Terry W Pearson, Andrew Pitt, Matthew E Pope, Andrew Porter, George W Preston, Morteza Razavi, Andrew Shapanis, Frances R Sidgwick, Raj Singh, Paul Skipp, Reynard Speiss, Justyna Spiewak, Anna L Tierney, Drupad Trivedi, Matthias Trost, Richard D Unwin, Luke Wainwright, Caitlin Walton-Doyle, Anthony D Whetton, Sandra Wilks, Richard Yip
Institutions: University of Southampton, University of Manchester, University of Liverpool, Guy's and St Thomas' NHS Foundation Trust, University of Reading, Evelina London Children's Healthcare, University College London, Newcastle University, Leicester Royal Infirmary, University of Leicester, Manchester University NHS Foundation Trust, University of Victoria, Cancer Research UK, University of Oxford, CRUK Lung Cancer Centre of Excellence
Language: English
Subfield: Infectious Diseases
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W4402137264