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A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen
ABSTRACT: To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)–specific monoclonal antibody (mAb), the Escherichia coli–expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Springer Berlin Heidelberg
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8236046/ https://www.ncbi.nlm.nih.gov/pubmed/34129082 http://dx.doi.org/10.1007/s00253-021-11342-7 |
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author | Kubickova, Barbara Schenk, Jörg A. Ramm, Franziska Markuškienė, Kornelija Reetz, Jochen Dremsek, Paul Tamosiunas, Paulius Lukas Cepulyte, Laima Trinh, Hoai Anh Scholz, Johannes Memczak, Henry Hovestädt, Marc Ryll, René Petraityte-Burneikiene, Rasa Corman, Victor M. Andersson, Anika Becher, Dietmar Groschup, Martin H. Kubick, Stefan Sellrie, Frank Johne, Reimar Ulrich, Rainer G. |
author_facet | Kubickova, Barbara Schenk, Jörg A. Ramm, Franziska Markuškienė, Kornelija Reetz, Jochen Dremsek, Paul Tamosiunas, Paulius Lukas Cepulyte, Laima Trinh, Hoai Anh Scholz, Johannes Memczak, Henry Hovestädt, Marc Ryll, René Petraityte-Burneikiene, Rasa Corman, Victor M. Andersson, Anika Becher, Dietmar Groschup, Martin H. Kubick, Stefan Sellrie, Frank Johne, Reimar Ulrich, Rainer G. |
author_sort | Kubickova, Barbara |
collection | PubMed |
description | ABSTRACT: To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)–specific monoclonal antibody (mAb), the Escherichia coli–expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli–expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient–derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes. KEY POINTS: • The antibody showed cross-reactivity with capsid proteins of different hepeviruses. • The linear epitope of the antibody was mapped in a partially surface-exposed region. • The antibody detected native HEV-3 antigen in infected mammalian cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-021-11342-7. |
format | Online Article Text |
id | pubmed-8236046 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-82360462021-07-09 A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen Kubickova, Barbara Schenk, Jörg A. Ramm, Franziska Markuškienė, Kornelija Reetz, Jochen Dremsek, Paul Tamosiunas, Paulius Lukas Cepulyte, Laima Trinh, Hoai Anh Scholz, Johannes Memczak, Henry Hovestädt, Marc Ryll, René Petraityte-Burneikiene, Rasa Corman, Victor M. Andersson, Anika Becher, Dietmar Groschup, Martin H. Kubick, Stefan Sellrie, Frank Johne, Reimar Ulrich, Rainer G. Appl Microbiol Biotechnol Biotechnological Products and Process Engineering ABSTRACT: To generate a hepatitis E virus (HEV) genotype 3 (HEV-3)–specific monoclonal antibody (mAb), the Escherichia coli–expressed carboxy-terminal part of its capsid protein was used to immunise BALB/c mice. The immunisation resulted in the induction of HEV-specific antibodies of high titre. The mAb G117-AA4 of IgG1 isotype was obtained showing a strong reactivity with the homologous E. coli, but also yeast-expressed capsid protein of HEV-3. The mAb strongly cross-reacted with ratHEV capsid protein derivatives produced in both expression systems and weaker with an E. coli–expressed batHEV capsid protein fragment. In addition, the mAb reacted with capsid protein derivatives of genotypes HEV-2 and HEV-4 and common vole hepatitis E virus (cvHEV), produced by the cell-free synthesis in Chinese hamster ovary (CHO) and Spodoptera frugiperda (Sf21) cell lysates. Western blot and line blot reactivity of the mAb with capsid protein derivatives of HEV-1 to HEV-4, cvHEV, ratHEV and batHEV suggested a linear epitope. Use of truncated derivatives of ratHEV capsid protein in ELISA, Western blot, and a Pepscan analysis allowed to map the epitope within a partially surface-exposed region with the amino acid sequence LYTSV. The mAb was also shown to bind to human patient–derived HEV-3 from infected cell culture and to hare HEV-3 and camel HEV-7 capsid proteins from transfected cells by immunofluorescence assay. The novel mAb may serve as a useful tool for further investigations on the pathogenesis of HEV infections and might be used for diagnostic purposes. KEY POINTS: • The antibody showed cross-reactivity with capsid proteins of different hepeviruses. • The linear epitope of the antibody was mapped in a partially surface-exposed region. • The antibody detected native HEV-3 antigen in infected mammalian cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00253-021-11342-7. Springer Berlin Heidelberg 2021-06-15 2021 /pmc/articles/PMC8236046/ /pubmed/34129082 http://dx.doi.org/10.1007/s00253-021-11342-7 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Biotechnological Products and Process Engineering Kubickova, Barbara Schenk, Jörg A. Ramm, Franziska Markuškienė, Kornelija Reetz, Jochen Dremsek, Paul Tamosiunas, Paulius Lukas Cepulyte, Laima Trinh, Hoai Anh Scholz, Johannes Memczak, Henry Hovestädt, Marc Ryll, René Petraityte-Burneikiene, Rasa Corman, Victor M. Andersson, Anika Becher, Dietmar Groschup, Martin H. Kubick, Stefan Sellrie, Frank Johne, Reimar Ulrich, Rainer G. A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title | A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title_full | A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title_fullStr | A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title_full_unstemmed | A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title_short | A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen |
title_sort | broadly cross-reactive monoclonal antibody against hepatitis e virus capsid antigen |
topic | Biotechnological Products and Process Engineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8236046/ https://www.ncbi.nlm.nih.gov/pubmed/34129082 http://dx.doi.org/10.1007/s00253-021-11342-7 |
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