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Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis
Hepatitis B virus (HBV) core antigen (HBc) is a structural protein that forms the viral nucleocapsid and is involved in various steps of the viral replication cycle, but its role in the pathogenesis of HBV infection is still elusive. In this study, we generated a mouse monoclonal antibody (mAb) agai...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9783060/ https://www.ncbi.nlm.nih.gov/pubmed/36557634 http://dx.doi.org/10.3390/microorganisms10122381 |
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author | Nakai, Yusuke Miyakawa, Kei Yamaoka, Yutaro Hatayama, Yasuyoshi Nishi, Mayuko Suzuki, Hidefumi Kimura, Hirokazu Takahashi, Hidehisa Kimura, Yayoi Ryo, Akihide |
author_facet | Nakai, Yusuke Miyakawa, Kei Yamaoka, Yutaro Hatayama, Yasuyoshi Nishi, Mayuko Suzuki, Hidefumi Kimura, Hirokazu Takahashi, Hidehisa Kimura, Yayoi Ryo, Akihide |
author_sort | Nakai, Yusuke |
collection | PubMed |
description | Hepatitis B virus (HBV) core antigen (HBc) is a structural protein that forms the viral nucleocapsid and is involved in various steps of the viral replication cycle, but its role in the pathogenesis of HBV infection is still elusive. In this study, we generated a mouse monoclonal antibody (mAb) against HBc and used it in antibody-based in situ biotinylation analysis in order to identify host proteins that interact with HBc. HBc antigen was produced with a wheat germ cell-free protein synthesis system and used to immunize mice. Among the established hybridoma clones, a single clone (mAb #7) was selected and further characterized for its ability in the antibody-based in situ biotinylation analysis to collect host proteins that are in the vicinity of HBc. Using mass spectrometry, we identified 215 HBc-interacting host proteins, three of which bind HBc most significantly under hypoxic conditions. Our results indicate that mAb #7 can be used to systematically identify host proteins that interact with HBc under pathophysiological conditions, and thus may be useful to explore the molecular pathways involved in HBV-induced cytopathogenesis. |
format | Online Article Text |
id | pubmed-9783060 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-97830602022-12-24 Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis Nakai, Yusuke Miyakawa, Kei Yamaoka, Yutaro Hatayama, Yasuyoshi Nishi, Mayuko Suzuki, Hidefumi Kimura, Hirokazu Takahashi, Hidehisa Kimura, Yayoi Ryo, Akihide Microorganisms Article Hepatitis B virus (HBV) core antigen (HBc) is a structural protein that forms the viral nucleocapsid and is involved in various steps of the viral replication cycle, but its role in the pathogenesis of HBV infection is still elusive. In this study, we generated a mouse monoclonal antibody (mAb) against HBc and used it in antibody-based in situ biotinylation analysis in order to identify host proteins that interact with HBc. HBc antigen was produced with a wheat germ cell-free protein synthesis system and used to immunize mice. Among the established hybridoma clones, a single clone (mAb #7) was selected and further characterized for its ability in the antibody-based in situ biotinylation analysis to collect host proteins that are in the vicinity of HBc. Using mass spectrometry, we identified 215 HBc-interacting host proteins, three of which bind HBc most significantly under hypoxic conditions. Our results indicate that mAb #7 can be used to systematically identify host proteins that interact with HBc under pathophysiological conditions, and thus may be useful to explore the molecular pathways involved in HBV-induced cytopathogenesis. MDPI 2022-11-30 /pmc/articles/PMC9783060/ /pubmed/36557634 http://dx.doi.org/10.3390/microorganisms10122381 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Nakai, Yusuke Miyakawa, Kei Yamaoka, Yutaro Hatayama, Yasuyoshi Nishi, Mayuko Suzuki, Hidefumi Kimura, Hirokazu Takahashi, Hidehisa Kimura, Yayoi Ryo, Akihide Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title | Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title_full | Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title_fullStr | Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title_full_unstemmed | Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title_short | Generation and Utilization of a Monoclonal Antibody against Hepatitis B Virus Core Protein for a Comprehensive Interactome Analysis |
title_sort | generation and utilization of a monoclonal antibody against hepatitis b virus core protein for a comprehensive interactome analysis |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9783060/ https://www.ncbi.nlm.nih.gov/pubmed/36557634 http://dx.doi.org/10.3390/microorganisms10122381 |
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