tetano
Editor, Senior Moderator
J Virol Methods
. 2024 Nov 16:115073.
doi: 10.1016/j.jviromet.2024.115073. Online ahead of print. Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants
Caroline L Ashley[SUP] 1 [/SUP], Malik Bloul[SUP] 2 [/SUP], Sibel Alca[SUP] 3 [/SUP], Lachlan Smith[SUP] 4 [/SUP], Wang Jin[SUP] 5 [/SUP], David Khoury[SUP] 6 [/SUP], Claudio Counoupas[SUP] 7 [/SUP], Miles Davenport[SUP] 8 [/SUP], James A Triccas[SUP] 9 [/SUP], Megan Steain[SUP] 10 [/SUP]
Affiliations
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[SUB]50[/SUB] 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[SUB]50[/SUB] 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.
Keywords: COVID-19; Neutralising antibodies; SARS-CoV-2; multiplex neutralisation assay; vaccine; variants.
. 2024 Nov 16:115073.
doi: 10.1016/j.jviromet.2024.115073. Online ahead of print. Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants
Caroline L Ashley[SUP] 1 [/SUP], Malik Bloul[SUP] 2 [/SUP], Sibel Alca[SUP] 3 [/SUP], Lachlan Smith[SUP] 4 [/SUP], Wang Jin[SUP] 5 [/SUP], David Khoury[SUP] 6 [/SUP], Claudio Counoupas[SUP] 7 [/SUP], Miles Davenport[SUP] 8 [/SUP], James A Triccas[SUP] 9 [/SUP], Megan Steain[SUP] 10 [/SUP]
Affiliations
- PMID: 39557342
- DOI: 10.1016/j.jviromet.2024.115073
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[SUB]50[/SUB] 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[SUB]50[/SUB] 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.
Keywords: COVID-19; Neutralising antibodies; SARS-CoV-2; multiplex neutralisation assay; vaccine; variants.