A flow cytometry-based assay to measure neutralizing antibodies against SARS-CoV-2 virus.
COVID‐19
PRNT
S protein
flow cytometry
Journal
Cytometry. Part A : the journal of the International Society for Analytical Cytology
ISSN: 1552-4930
Titre abrégé: Cytometry A
Pays: United States
ID NLM: 101235694
Informations de publication
Date de publication:
16 Apr 2024
16 Apr 2024
Historique:
revised:
11
03
2024
received:
25
09
2023
accepted:
25
03
2024
medline:
16
4
2024
pubmed:
16
4
2024
entrez:
16
4
2024
Statut:
aheadofprint
Résumé
The COVID-19 pandemic caused by the SARS-CoV-2 virus has highlighted the need for serological assays that can accurately evaluate the neutralizing efficiency of antibodies produced during infection or induced by vaccines. However, conventional assays often require the manipulation of live viruses on a level-three biosafety (BSL3) facility, which presents practical and safety challenges. Here, we present a novel, alternative assay that measures neutralizing antibodies (NAbs) against SARS-CoV-2 in plasma using flow cytometry. This assay is based on antibody binding to the S protein and has demonstrated precision in both intra- and inter-assay measurements at a dilution of 1:50. The cut-off was determined using Receiver Operating Characteristic (ROC) analysis and the value of 36.01% has shown high sensitivity and specificity in distinguishing between pre-pandemic sera, COVID-19 patients, and vaccinated individuals. The efficiency significantly correlates with the gold standard test, PRNT. Our new assay offers a safe and efficient alternative to conventional assays for evaluating NAbs against SARS-CoV-2.
Identifiants
pubmed: 38624015
doi: 10.1002/cyto.a.24838
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Coordenação de Aperfeiçoamento de Pessoal de Nível superior - CAPES
Organisme : Brazilian Ministry of Health (PROADI)
Organisme : Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq
Informations de copyright
© 2024 International Society for Advancement of Cytometry.
Références
Chan JFW, Yuan S, Kok KH, To KKW, Chu H, Yang J, et al. A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person‐to‐person transmission: a study of a family cluster. Lancet. 2020;395:514–523.
Zhou P, Yang XL, Wang XG, Hu B, Zhang L, Zhang W, et al. A pneumonia outbreak associated with a new coronavirus of probable bat origin. Nature. 2020;579:270–273.
WHO. WHO Director‐General's opening remarks at the media briefing on COVID‐19 – 11 March 2020. 2020.
Hu Y, Wen J, Tang L, Zhang H, Zhang X, Li Y, et al. The M protein of SARS‐CoV: basic structural and immunological properties. Genom Proteom Bioinformat/Beijing Genom Inst. 2003;1:118–130.
Wu F, Wang A, Liu M, Wang Q, Chen J, Xia S, et al. Neutralizing antibody responses to SARS‐CoV‐2 in a COVID‐19 recovered patient cohort and their implications. SSRN Electron J. 2020;152(1&2):82‐87.
Wu J, Liang B, Chen C, Wang H, Fang Y, Shen S, et al. SARS‐CoV‐2 infection induces sustained humoral immune responses in convalescent patients following symptomatic COVID‐19. Nat Commun. 2021;12:1.
He Y, Zhou Y, Liu S, Kou Z, Li W, Farzan M, et al. Receptor‐binding domain of SARS‐CoV spike protein induces highly potent NAbs: implication for developing subunit vaccine. Biochem Biophys Res Commun. 2004;324:773–781.
He Y, Lu H, Siddiqui P, Zhou Y, Jiang S. Receptor‐binding domain of severe acute respiratory syndrome coronavirus spike protein contains multiple conformation‐dependent epitopes that induce highly potent NAbs. J Immunol. 2005;174:4908–4915.
Yu F, Xiang R, Deng X, Wang L, Yu Z, Tian S, et al. Receptor‐binding domain‐specific human neutralizing monoclonal antibodies against SARS‐CoV and SARS‐CoV‐2. Signal transduction and targeted therapy. 2020;5(1):212.
Krammer F. SARS‐CoV‐2 vaccines in development. Nature. 2020;586:516–527.
Kyriakidis NC, López‐Cortés A, González EV, Grimaldos AB, Prado EO. SARS‐CoV‐2 vaccines strategies: a comprehensive review of phase 3 candidates. NPJ Vaccines. 2021;6(1):28.
Samrat SK, Tharappel AM, Li Z, Li H. Prospect of SARS‐CoV‐2 spike protein: potential role in vaccine and therapeutic development. Virus research. 2020.
Muruato AE, Fontes‐Garfias CR, Ren P, Garcia‐Blanco MA, Menachery VD, Xie X, et al. A high‐throughput neutralizing antibody assay for COVID‐19 diagnosis and vaccine evaluation. Nat Commun. 2020;11(1):4059.
Kraus AA, Messer W, Haymore LB, De Silva AM. Comparison of plaque‐ and flow cytometry‐based methods for measuring dengue virus neutralization. J Clin Microbiol. 2007;45:3777–3780.
Bordignon J, Comin F, Ferreira SC, Caporale GM, Lima Filho JH, Zanetti CR. Calculating rabies virus neutralizing antibodies titres by flow cytometry. Rev Inst Med Trop Sao Paulo. 2002;44(3):151–154. doi:10.1590/s0036‐46652002000300007
Tan CW, Chia WN, Qin X, Liu P, Chen MIC, Tiu C, et al. A SARS‐CoV‐2 surrogate virus neutralization test based on antibody‐mediated blockage of ACE2–spike protein–protein interaction. Nat Biotechnol. 2020;38:1073–1078.
Abe KT, Li Z, Samson R, Samavarchi‐Tehrani P, Valcourt EJ, Wood H, et al. A simple protein‐based surrogate neutralization assay for SARS‐CoV‐2. JCI Insight. 2020;5:1.
Horndler L, Delgado P, Abia D, Balabanov I, Martínez‐Fleta P, Cornish G, et al. Flow cytometry multiplexed method for the detection of neutralizing human antibodies to the native SARS‐CoV‐2 spike protein. EMBO Mol Med. 2021;13:e13549.
Gai J, Ma L, Li G, Zhu M, Qiao P, Li X, et al. A potent neutralizing nanobody against SARS‐CoV‐2 with inhaled delivery potential. MedComm. 2021;2:101–113.
Amanat F, Stadlbauer D, Strohmeier S, Nguyen THO, Chromikova V, McMahon M, et al. A serological assay to detect SARS‐CoV‐2 seroconversion in humans. Nat Med. 2020;26:1033–1036. https://doi.org/10.1038/s41591-020-0913-5
Fazolo T, Lima K, Fontoura JC, de Souza PO, Hilario G, Zorzetto R, et al. Pediatric COVID‐19 patients in South Brazil show abundant viral mRNA and strong specific anti‐viral responses. Nat Commun. 2021;12:6844. https://doi.org/10.1038/s41467-021-27120-y
Souza WM, Amorim MR, Sesti‐Costa R, Coimbra LD, Brunetti NS, Toledo‐Teixeira DA, et al. Neutralisation of SARS‐CoV‐2 lineage P.1 by antibodies elicited through natural SARS‐CoV‐2 infection or vaccination with an inactivated SARS‐CoV‐2 vaccine: an immunological study. Lancet. 2021;2(10):e527–e535.
Tian L, Elsheikh EB, Patrone PN, Kearsley AJ, Gaigalas AK, Inwood S, et al. Article towards quantitative and standardized serological and neutralization assays for COVID‐19. Int J Mol Sci. 2021;22(5):1–15.
Egia‐Mendikute L, Bosch A, Prieto‐Fernández E, Vila‐Vecilla L, Zanetti SR, Lee SY, et al. A flow cytometry‐based neutralization assay for simultaneous evaluation of blocking antibodies against SARS‐CoV‐2 variants. Front Immunol. 2022;13:1014309. https://doi.org/10.3389/fimmu.2022.1014309
Reed GF, Lynn F, Meade BD. Use of relative standard deviation in assessing variability of quantitative assays. Clin Diagn Lab Immunol. 2002;9:1235–1239.