Cargando…

Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay

ABSTRACT: The nucleic acid test is still the standard assessment for the diagnosis of coronavirus disease 2019 (COVID-19), which is caused by human infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In addition to supporting the confirmation of disease cases, serological assa...

Descripción completa

Detalles Bibliográficos
Autores principales: Hsiao, Chia-Chun, Chiang, Yi-Wei, Chao, Tai-Ling, Tsai, Zen-Uong, Wang, Ting-Xuan, Jiang, Yu-Wei, Hsu, Hsiang-Fu, Lu, De-Chao, Wang, Jann-Tay, Wang, Jen-Ren, Wang, An-Bang, Chang, Sui-Yuan, Chang, Shih-Chung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9676789/
https://www.ncbi.nlm.nih.gov/pubmed/36404356
http://dx.doi.org/10.1007/s00253-022-12288-0
_version_ 1784833674015932416
author Hsiao, Chia-Chun
Chiang, Yi-Wei
Chao, Tai-Ling
Tsai, Zen-Uong
Wang, Ting-Xuan
Jiang, Yu-Wei
Hsu, Hsiang-Fu
Lu, De-Chao
Wang, Jann-Tay
Wang, Jen-Ren
Wang, An-Bang
Chang, Sui-Yuan
Chang, Shih-Chung
author_facet Hsiao, Chia-Chun
Chiang, Yi-Wei
Chao, Tai-Ling
Tsai, Zen-Uong
Wang, Ting-Xuan
Jiang, Yu-Wei
Hsu, Hsiang-Fu
Lu, De-Chao
Wang, Jann-Tay
Wang, Jen-Ren
Wang, An-Bang
Chang, Sui-Yuan
Chang, Shih-Chung
author_sort Hsiao, Chia-Chun
collection PubMed
description ABSTRACT: The nucleic acid test is still the standard assessment for the diagnosis of coronavirus disease 2019 (COVID-19), which is caused by human infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In addition to supporting the confirmation of disease cases, serological assays are used for the analysis of antibody status and epidemiological surveys. In this study, a single Western blot strip (WBS) coated with multiple Escherichia coli (E. coli)-expressed SARS-CoV-2 antigens was developed for comprehensive studies of antibody profiles in COVID-19 patient sera. The levels of specific antibodies directed to SARS-CoV-2 spike (S), S2, and nucleocapsid (N) proteins were gradually increased with the same tendency as the disease progressed after hospitalization. The signal readouts of S, S2, and N revealed by the multi-antigen-coated WBS (mWBS)-based serological assay (mWBS assay) also demonstrated a positive correlation with the SARS-CoV-2 neutralizing potency of the sera measured by the plaque reduction neutralization test (PRNT) assays. Surprisingly, the detection signals against the unstructured receptor-binding domain (RBD) purified from E. coli inclusion bodies were not observed, although the COVID-19 patient sera exhibited strong neutralizing potency in the PRNT assays, suggesting that the RBD-specific antibodies in patient sera mostly recognize the conformational epitopes. Furthermore, the mWBS assay identified a unique and major antigenic epitope at the residues 1148, 1149, 1152, 1155, and 1156 located within the 1127–1167 fragment of the S2 subunit, which was specifically recognized by the COVID-19 patient serum. The mWBS assay can be finished within 14–16 min by using the automatic platform of Western blotting by thin-film direct coating with suction (TDCS WB). Collectively, the mWBS assay can be applied for the analysis of antibody responses, prediction of the protective antibody status, and identification of the specific epitope. KEY POINTS: • A Western blot strip (WBS) coated with multiple SARS-CoV-2 antigens was developed for the serological assay. • The multi-antigen-coated WBS (mWBS) can be utilized for the simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens. • The mWBS-based serological assay (mWBS assay) identified a unique epitope recognized by the COVID-19 patient serum.
format Online
Article
Text
id pubmed-9676789
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Springer Berlin Heidelberg
record_format MEDLINE/PubMed
spelling pubmed-96767892022-11-21 Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay Hsiao, Chia-Chun Chiang, Yi-Wei Chao, Tai-Ling Tsai, Zen-Uong Wang, Ting-Xuan Jiang, Yu-Wei Hsu, Hsiang-Fu Lu, De-Chao Wang, Jann-Tay Wang, Jen-Ren Wang, An-Bang Chang, Sui-Yuan Chang, Shih-Chung Appl Microbiol Biotechnol Applied Genetics and Molecular Biotechnology ABSTRACT: The nucleic acid test is still the standard assessment for the diagnosis of coronavirus disease 2019 (COVID-19), which is caused by human infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In addition to supporting the confirmation of disease cases, serological assays are used for the analysis of antibody status and epidemiological surveys. In this study, a single Western blot strip (WBS) coated with multiple Escherichia coli (E. coli)-expressed SARS-CoV-2 antigens was developed for comprehensive studies of antibody profiles in COVID-19 patient sera. The levels of specific antibodies directed to SARS-CoV-2 spike (S), S2, and nucleocapsid (N) proteins were gradually increased with the same tendency as the disease progressed after hospitalization. The signal readouts of S, S2, and N revealed by the multi-antigen-coated WBS (mWBS)-based serological assay (mWBS assay) also demonstrated a positive correlation with the SARS-CoV-2 neutralizing potency of the sera measured by the plaque reduction neutralization test (PRNT) assays. Surprisingly, the detection signals against the unstructured receptor-binding domain (RBD) purified from E. coli inclusion bodies were not observed, although the COVID-19 patient sera exhibited strong neutralizing potency in the PRNT assays, suggesting that the RBD-specific antibodies in patient sera mostly recognize the conformational epitopes. Furthermore, the mWBS assay identified a unique and major antigenic epitope at the residues 1148, 1149, 1152, 1155, and 1156 located within the 1127–1167 fragment of the S2 subunit, which was specifically recognized by the COVID-19 patient serum. The mWBS assay can be finished within 14–16 min by using the automatic platform of Western blotting by thin-film direct coating with suction (TDCS WB). Collectively, the mWBS assay can be applied for the analysis of antibody responses, prediction of the protective antibody status, and identification of the specific epitope. KEY POINTS: • A Western blot strip (WBS) coated with multiple SARS-CoV-2 antigens was developed for the serological assay. • The multi-antigen-coated WBS (mWBS) can be utilized for the simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens. • The mWBS-based serological assay (mWBS assay) identified a unique epitope recognized by the COVID-19 patient serum. Springer Berlin Heidelberg 2022-11-21 2022 /pmc/articles/PMC9676789/ /pubmed/36404356 http://dx.doi.org/10.1007/s00253-022-12288-0 Text en © The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.
spellingShingle Applied Genetics and Molecular Biotechnology
Hsiao, Chia-Chun
Chiang, Yi-Wei
Chao, Tai-Ling
Tsai, Zen-Uong
Wang, Ting-Xuan
Jiang, Yu-Wei
Hsu, Hsiang-Fu
Lu, De-Chao
Wang, Jann-Tay
Wang, Jen-Ren
Wang, An-Bang
Chang, Sui-Yuan
Chang, Shih-Chung
Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title_full Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title_fullStr Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title_full_unstemmed Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title_short Simultaneous detection of antibody responses to multiple SARS-CoV-2 antigens by a Western blot serological assay
title_sort simultaneous detection of antibody responses to multiple sars-cov-2 antigens by a western blot serological assay
topic Applied Genetics and Molecular Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9676789/
https://www.ncbi.nlm.nih.gov/pubmed/36404356
http://dx.doi.org/10.1007/s00253-022-12288-0
work_keys_str_mv AT hsiaochiachun simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT chiangyiwei simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT chaotailing simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT tsaizenuong simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT wangtingxuan simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT jiangyuwei simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT hsuhsiangfu simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT ludechao simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT wangjanntay simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT wangjenren simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT wanganbang simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT changsuiyuan simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay
AT changshihchung simultaneousdetectionofantibodyresponsestomultiplesarscov2antigensbyawesternblotserologicalassay