Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants.

Autor: Ashley CL; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia., Bloul M; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: malik.i.bloul@gmail.com., Alca S; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: sibel.alca@sydney.edu.au., Smith L; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: lachlan.smith1@sydney.edu.au., Jin W; Kirby Institute, University of New South Wales, Sydney, NSW, 2052, Australia. Electronic address: jwang@kirby.unsw.edu.au., Khoury D; Kirby Institute, University of New South Wales, Sydney, NSW, 2052, Australia. Electronic address: dkhoury@kirby.unsw.edu.au., Counoupas C; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia; Centre for Infection and Immunity, Centenary Institute, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: claudio.counoupas@sydney.edu.au., Davenport M; Kirby Institute, University of New South Wales, Sydney, NSW, 2052, Australia. Electronic address: M.Davenport@unsw.edu.au., Triccas JA; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia; Centre for Infection and Immunity, Centenary Institute, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: jamie.triccas@sydney.edu.au., Steain M; Sydney Infectious Diseases Institute (Sydney ID), Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW 2006, Australia; Charles Perkin Centre, The University of Sydney, Camperdown, NSW 2006, Australia. Electronic address: megan.steain@sydney.edu.au.
Jazyk: angličtina
Zdroj: Journal of virological methods [J Virol Methods] 2025 Feb; Vol. 332, pp. 115073. Date of Electronic Publication: 2024 Nov 16.
DOI: 10.1016/j.jviromet.2024.115073
Abstrakt: A multiplexed, lentivirus-based pseudovirus neutralisation assay (pVNT) was developed for high-throughput measurement of neutralising antibodies (nAbs) against three distinct SARS-CoV-2 spike variants. Intra-assay variability was minimised by optimising the plate layout and determining an optimal percentage transduction for the pseudovirus inoculum. Comparison of EC 50 titres between single and multiplexed pVNT assays showed no significant differences, indicating reliability of the multiplexed assay. Evaluation of convalescent human sera confirmed assay robustness, with consistent EC 50 titres for variant pseudoviruses relative to the ancestral strain observed across single and multiplexed assays. This multiplexed pVNT provides a reliable tool for assessing nAb responses against SARS-CoV-2 variants and could be used to accelerate preclinical vaccine assessment in preparation for the next coronavirus pandemic.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
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