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A highly sensitive ...
A highly sensitive bead-based flow cytometric competitive binding assay to detect SARS-CoV-2 neutralizing antibody activity
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- Yao, Xiangyu (författare)
- Shanxi University
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- Zhang, Zhichao (författare)
- Shanxi Academy of Advanced Research and Innovation
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- Mei, Qingmin (författare)
- University of Science and Technology of China
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- Li, Shenwei (författare)
- Shanghai International Travel Healthcare Center
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- Xing, Li (författare)
- Shanxi Provincial Key Laboratory for Major Infectious Disease Response,Shanxi University
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- Long, Yali (författare)
- Shanxi University
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- Zhang, Demei (författare)
- Taiyuan Blood Center
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- Wang, Jing (författare)
- University of Science and Technology of China
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- Wang, Xiedong (författare)
- Shanxi University
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- Xie, Bin (författare)
- Lund University,Lunds universitet,Tillämpad biokemi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Pure and Applied Biochemistry,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH
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- Yang, Bo (författare)
- Shanxi Academy of Advanced Research and Innovation,Shanxi Provincial Key Laboratory for Major Infectious Disease Response
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- Gao, Yong (författare)
- University of Science and Technology of China
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- Wu, Changxin (författare)
- Shanxi Provincial Key Laboratory for Major Infectious Disease Response,Shanxi University
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- Meng, Qinglai (författare)
- Shanxi University,Shanxi Provincial Key Laboratory for Major Infectious Disease Response
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(creator_code:org_t)
- 2022-11-30
- 2022
- Engelska 15 s.
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Ingår i: Frontiers in Immunology. - : Frontiers Media SA. - 1664-3224. ; 13
- Relaterad länk:
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http://dx.doi.org/10... (free)
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https://lup.lub.lu.s...
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https://doi.org/10.3...
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Abstract
Ämnesord
Stäng
- Accurate detection of SARS-CoV-2 neutralizing antibody (nAb) is critical for assessing the immunity levels after virus infection or vaccination. As fast, cost-effective alternatives to viral infection-based assays, competitive binding (CB) assays were developed to quantitate nAb by monitoring the ability of sera to inhibit the binding of viral spike (S) protein to the angiotensin converting enzyme 2 (ACE2) receptor. Herein, we established a bead-based flow cytometric CB assay and tested the detection performance of six combination models, i.e. immobilized ACE2 and soluble Fc-tagged S1 subunit of S protein (iACE2/S1-Fc), immobilized ACE2 and soluble Fc-tagged receptor binding domain (RBD) of S protein (iACE2/RBD-Fc), immobilized S1 and soluble Fc-tagged ACE2 (iS1/ACE2-Fc), immobilized S1 and soluble His-tagged ACE2 (iS1/ACE2-His), immobilized RBD and soluble Fc-tagged ACE2 (iRBD/ACE2-Fc), and immobilized RBD and soluble His-tagged ACE2 (iRBD/ACE2-His). Using SARS-CoV-2 monoclonal antibodies and sera of convalescent COVID-19 patients and vaccinated subjects, the combination models iACE2/RBD-Fc, iACE2/S1-Fc and iS1/ACE2-His were identified to be able to specifically detect SARS-CoV-2 nAb, among which iACE2/RBD-Fc model showed the highest sensitivity, superior to a commercial SARS-CoV-2 surrogate virus neutralization test (sVNT) ELISA kit. Further studies demonstrated that the sensitivity and specificity of CB assays were affected by the tag of ACE2, type of spike and method of measuring binding rate between ACE2 and spike. Moreover, the iACE2/RBD-Fc model showed good performance in detecting kinetic development of nAb against both the prototype SARS-CoV-2 strain and an omicron variant of SARS-CoV-2 in people immunized by an inactivated SARS-CoV-2 vaccine, and the results of iACE2/RBD-Fc model are correlated well with those of live virus-based and pseudovirus-based neutralization tests, demonstrating the potential to be developed into a highly sensitive, specific, versatile and high-throughput method for detecting SARS-CoV-2 nAb in clinical practice.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Immunologi inom det medicinska området (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Immunology in the medical area (hsv//eng)
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Yao, Xiangyu
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Zhang, Zhichao
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Mei, Qingmin
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Li, Shenwei
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Xing, Li
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Long, Yali
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visa fler...
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Zhang, Demei
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Wang, Jing
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Wang, Xiedong
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Xie, Bin
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Yang, Bo
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Gao, Yong
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Wu, Changxin
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Meng, Qinglai
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