A High-Throughput Broad Neutralizing Antibody Assay for Detecting SARS-CoV-2 Variant Immunity in Population.

IF 3.8 2区医学 Q2 CHEMISTRY, MEDICINAL

ACS Infectious Diseases Pub Date : 2025-09-12 DOI:10.1021/acsinfecdis.5c00402

Xiaohan Zhang, Yajie Wang, Mansheng Li, Haolong Li, Xiaomei Zhang, Xingming Xu, Qingqing Ma, Di Hu, Yan Jia, Te Liang, Yunping Zhu, Surbhi Bihani, Sanjeeva Srivastava, Manuel Fuentes, Yongzhe Li, Xiaoxu Han, Bingwei Wang, Xiaobo Yu

{"title":"A High-Throughput Broad Neutralizing Antibody Assay for Detecting SARS-CoV-2 Variant Immunity in Population.","authors":"Xiaohan Zhang, Yajie Wang, Mansheng Li, Haolong Li, Xiaomei Zhang, Xingming Xu, Qingqing Ma, Di Hu, Yan Jia, Te Liang, Yunping Zhu, Surbhi Bihani, Sanjeeva Srivastava, Manuel Fuentes, Yongzhe Li, Xiaoxu Han, Bingwei Wang, Xiaobo Yu","doi":"10.1021/acsinfecdis.5c00402","DOIUrl":null,"url":null,"abstract":"Detecting neutralizing antibodies (NAbs) to SARS-CoV-2 variants that are evolved is crucial to know the escape of host immunity to the newly arising variants. To address this need, we developed a high-throughput broad neutralizing antibody (bNAb) assay using flow cytometry with magnetic-fluorescent microspheres for detecting NAbs against diverse SARS-CoV-2 variants. The assay is rapid, reliable, and 35-fold more sensitive than Luminex technology. Our results highly correlated with the IgG serological assay (R = 0.90), the FDA-approved cPass sVNT assay (R = 0.92), pseudovirus-based neutralization assay (R = 0.96, 0.66, 0.65), and live virus-based neutralization assay (R = 0.79, 0.64). When applied to 56 healthy individuals receiving third-dose vaccines (18 CoronaVac; 38 ZF2001) and 35 HIV patients with breakthrough infection of COVID-19 (16 with CD4+ < 350 cells/μL; 19 with CD4+ > 500 cells/μL), results showed that the Omicron BA.1-BA.5 variants exhibited significant resistance to inactivated vaccines in healthy individuals. In HIV patients, the breakthrough infection of Omicron BA.5.2 or BF.7 variants can induce broad neutralizing activity to non-Omicron and Omicron variants together with vaccination. Notably, the levels of NAbs against most of the SARS-CoV-2 variants are much lower in the decreased immunity of HIV patients (CD4+ < 350 cells/μL) compared to the recovered immunity (CD4+ > 500 cells/μL), indicating that maintenance of the immune system is crucial for NAb production. Altogether, our high-throughput proteomics platform represents a powerful tool for the detection of bNAbs in the population and may inform the development of more effective COVID-19 vaccines and vaccination strategies in the future.","PeriodicalId":17,"journal":{"name":"ACS Infectious Diseases","volume":" ","pages":""},"PeriodicalIF":3.8000,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Infectious Diseases","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1021/acsinfecdis.5c00402","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, MEDICINAL","Score":null,"Total":0}

引用次数: 0

Abstract

Detecting neutralizing antibodies (NAbs) to SARS-CoV-2 variants that are evolved is crucial to know the escape of host immunity to the newly arising variants. To address this need, we developed a high-throughput broad neutralizing antibody (bNAb) assay using flow cytometry with magnetic-fluorescent microspheres for detecting NAbs against diverse SARS-CoV-2 variants. The assay is rapid, reliable, and 35-fold more sensitive than Luminex technology. Our results highly correlated with the IgG serological assay (R = 0.90), the FDA-approved cPass sVNT assay (R = 0.92), pseudovirus-based neutralization assay (R = 0.96, 0.66, 0.65), and live virus-based neutralization assay (R = 0.79, 0.64). When applied to 56 healthy individuals receiving third-dose vaccines (18 CoronaVac; 38 ZF2001) and 35 HIV patients with breakthrough infection of COVID-19 (16 with CD4+ < 350 cells/μL; 19 with CD4+ > 500 cells/μL), results showed that the Omicron BA.1-BA.5 variants exhibited significant resistance to inactivated vaccines in healthy individuals. In HIV patients, the breakthrough infection of Omicron BA.5.2 or BF.7 variants can induce broad neutralizing activity to non-Omicron and Omicron variants together with vaccination. Notably, the levels of NAbs against most of the SARS-CoV-2 variants are much lower in the decreased immunity of HIV patients (CD4+ < 350 cells/μL) compared to the recovered immunity (CD4+ > 500 cells/μL), indicating that maintenance of the immune system is crucial for NAb production. Altogether, our high-throughput proteomics platform represents a powerful tool for the detection of bNAbs in the population and may inform the development of more effective COVID-19 vaccines and vaccination strategies in the future.

查看原文本刊更多论文

高通量广谱中和抗体检测人群SARS-CoV-2变异免疫

检测进化出的针对SARS-CoV-2变体的中和抗体（nab）对于了解宿主对新出现的变体的免疫逃逸至关重要。为了满足这一需求，我们开发了一种高通量宽中和抗体（bNAb）检测方法，使用磁荧光微球流式细胞术检测不同SARS-CoV-2变体的nab。该方法快速、可靠，灵敏度是Luminex技术的35倍。我们的结果与IgG血清学检测（R = 0.90）、fda批准的cPass sVNT检测（R = 0.92）、基于假病毒的中和检测（R = 0.96、0.66、0.65）和基于活病毒的中和检测（R = 0.79、0.64）高度相关。对56例接受第三剂疫苗的健康人（CoronaVac 18例，ZF2001 38例）和35例突破感染的新冠病毒感染者（CD4+ < 350细胞/μL 16例，CD4+ > 500细胞/μL 19例）进行分析，结果表明，Omicron BA.1-BA在不同浓度下的表达量不同。5个变异在健康个体中对灭活疫苗表现出显著的耐药性。在HIV患者中，Omicron BA.5.2或BF.7变体的突破感染可以诱导对非Omicron和Omicron变体的广泛中和活性，并结合疫苗接种。值得注意的是，在HIV患者免疫力下降（CD4+ < 350细胞/μL）时，针对大多数SARS-CoV-2变体的NAb水平远低于恢复免疫力（CD4+ > 500细胞/μL），这表明免疫系统的维持对NAb的产生至关重要。总之，我们的高通量蛋白质组学平台是检测人群中bnab的强大工具，可能为未来开发更有效的COVID-19疫苗和疫苗接种策略提供信息。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

ACS Infectious Diseases CHEMISTRY, MEDICINALINFECTIOUS DISEASES&nb-INFECTIOUS DISEASES

CiteScore

9.70

自引率

3.80%

发文量

213

期刊介绍： ACS Infectious Diseases will be the first journal to highlight chemistry and its role in this multidisciplinary and collaborative research area. The journal will cover a diverse array of topics including, but not limited to: * Discovery and development of new antimicrobial agents — identified through target- or phenotypic-based approaches as well as compounds that induce synergy with antimicrobials. * Characterization and validation of drug target or pathways — use of single target and genome-wide knockdown and knockouts, biochemical studies, structural biology, new technologies to facilitate characterization and prioritization of potential drug targets. * Mechanism of drug resistance — fundamental research that advances our understanding of resistance; strategies to prevent resistance. * Mechanisms of action — use of genetic, metabolomic, and activity- and affinity-based protein profiling to elucidate the mechanism of action of clinical and experimental antimicrobial agents. * Host-pathogen interactions — tools for studying host-pathogen interactions, cellular biochemistry of hosts and pathogens, and molecular interactions of pathogens with host microbiota. * Small molecule vaccine adjuvants for infectious disease. * Viral and bacterial biochemistry and molecular biology.