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Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody
A swine acute diarrhea syndrome coronavirus (SADS-CoV) that causes severe diarrhea in suckling piglets was identified in Southern China in 2017. To develop an antigen that is specific, sensitive, and easy to prepare for serological diagnosis, antigenic sites in the SADS-CoV nucleocapsid (N) protein...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114574/ https://www.ncbi.nlm.nih.gov/pubmed/31518629 http://dx.doi.org/10.1016/j.virusres.2019.197752 |
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author | Han, Yuru Zhang, Jiyu Shi, Hongyan Zhou, Ling Chen, Jianfei Zhang, Xin Liu, Jianbo Zhang, Jialin Wang, Xiaobo Ji, Zhaoyang Jing, Zhaoyang Cong, Guangyi Ma, Jingyun Shi, Da Li, Feng |
author_facet | Han, Yuru Zhang, Jiyu Shi, Hongyan Zhou, Ling Chen, Jianfei Zhang, Xin Liu, Jianbo Zhang, Jialin Wang, Xiaobo Ji, Zhaoyang Jing, Zhaoyang Cong, Guangyi Ma, Jingyun Shi, Da Li, Feng |
author_sort | Han, Yuru |
collection | PubMed |
description | A swine acute diarrhea syndrome coronavirus (SADS-CoV) that causes severe diarrhea in suckling piglets was identified in Southern China in 2017. To develop an antigen that is specific, sensitive, and easy to prepare for serological diagnosis, antigenic sites in the SADS-CoV nucleocapsid (N) protein were screened. We generated and characterized an N-reactive monoclonal antibody (mAb) 3E9 from mice immunized with recombinant N protein. Through fine epitope mapping of mAb 3E9 using a panel of eukaryotic-expressed polypeptides with GFP-tags, we identified the motif (343)DAPVFTPAP(351) as the minimal unit of the linear B-cell epitope recognized by mAb 3E9. Protein sequence alignment indicated that (343)DAPVFTPAP(351) was highly conserved in different SADS-CoV strains and SADS-related coronaviruses from bat, with one substitution in this motif in HKU2-related bat coronavirus. Using mAb 3E9, we observed that N protein was expressed in the cytoplasm and was in the nucleolus during SADS-CoV replication. N protein was immunoprecipitated from SADS-CoV-infected Vero E6 cells. Taken together, our results indicated that 3E9 mAb could be a useful tool to investigate the structure and function of N protein during viral replication. |
format | Online Article Text |
id | pubmed-7114574 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71145742020-04-02 Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody Han, Yuru Zhang, Jiyu Shi, Hongyan Zhou, Ling Chen, Jianfei Zhang, Xin Liu, Jianbo Zhang, Jialin Wang, Xiaobo Ji, Zhaoyang Jing, Zhaoyang Cong, Guangyi Ma, Jingyun Shi, Da Li, Feng Virus Res Article A swine acute diarrhea syndrome coronavirus (SADS-CoV) that causes severe diarrhea in suckling piglets was identified in Southern China in 2017. To develop an antigen that is specific, sensitive, and easy to prepare for serological diagnosis, antigenic sites in the SADS-CoV nucleocapsid (N) protein were screened. We generated and characterized an N-reactive monoclonal antibody (mAb) 3E9 from mice immunized with recombinant N protein. Through fine epitope mapping of mAb 3E9 using a panel of eukaryotic-expressed polypeptides with GFP-tags, we identified the motif (343)DAPVFTPAP(351) as the minimal unit of the linear B-cell epitope recognized by mAb 3E9. Protein sequence alignment indicated that (343)DAPVFTPAP(351) was highly conserved in different SADS-CoV strains and SADS-related coronaviruses from bat, with one substitution in this motif in HKU2-related bat coronavirus. Using mAb 3E9, we observed that N protein was expressed in the cytoplasm and was in the nucleolus during SADS-CoV replication. N protein was immunoprecipitated from SADS-CoV-infected Vero E6 cells. Taken together, our results indicated that 3E9 mAb could be a useful tool to investigate the structure and function of N protein during viral replication. Elsevier B.V. 2019-11 2019-09-10 /pmc/articles/PMC7114574/ /pubmed/31518629 http://dx.doi.org/10.1016/j.virusres.2019.197752 Text en © 2019 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Han, Yuru Zhang, Jiyu Shi, Hongyan Zhou, Ling Chen, Jianfei Zhang, Xin Liu, Jianbo Zhang, Jialin Wang, Xiaobo Ji, Zhaoyang Jing, Zhaoyang Cong, Guangyi Ma, Jingyun Shi, Da Li, Feng Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title | Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title_full | Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title_fullStr | Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title_full_unstemmed | Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title_short | Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
title_sort | epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114574/ https://www.ncbi.nlm.nih.gov/pubmed/31518629 http://dx.doi.org/10.1016/j.virusres.2019.197752 |
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