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Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein

The transmissible gastroenteritis virus (TGEV) nucleocapsid (N) protein plays important roles in the replication and translation of viral RNA. The present study provides the first description of two monoclonal antibodies (mAbs) (5E8 and 3D7) directed against the TGEV N protein linker region (LKR) an...

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Autores principales: Zhang, Xin, Zhao, Xin, Dong, Hui, Zhu, Yunnuan, Shi, Hongyan, Chen, Jianfei, Shi, Da, Feng, Li
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5045181/
https://www.ncbi.nlm.nih.gov/pubmed/27689694
http://dx.doi.org/10.1371/journal.pone.0163920
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author Zhang, Xin
Zhao, Xin
Dong, Hui
Zhu, Yunnuan
Shi, Hongyan
Chen, Jianfei
Shi, Da
Feng, Li
author_facet Zhang, Xin
Zhao, Xin
Dong, Hui
Zhu, Yunnuan
Shi, Hongyan
Chen, Jianfei
Shi, Da
Feng, Li
author_sort Zhang, Xin
collection PubMed
description The transmissible gastroenteritis virus (TGEV) nucleocapsid (N) protein plays important roles in the replication and translation of viral RNA. The present study provides the first description of two monoclonal antibodies (mAbs) (5E8 and 3D7) directed against the TGEV N protein linker region (LKR) and carboxyl terminal domain (CTD). The mAbs 5E8 and 3D7 reacted with native N protein in western blotting and immunofluorescence assay (IFA). Two linear epitopes, (189)SVEQAVLAALKKLG(202) and (246)VTRFYGARSSSA(257), located in the LKR and CTD of TGEV N protein, respectively, were identified after truncating the protein and applying a peptide scanning technique. Using mAb 5E8, we observed that the N protein was expressed in the cytoplasm during TGEV replication and that the protein could be immunoprecipitated from TGEV-infected PK-15 cells. The mAb 5E8 can be applied for different approaches to diagnosis of TGEV infection. In addition, the antibodies represent useful tools for investigating the antigenic properties of the N protein.
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spelling pubmed-50451812016-10-27 Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein Zhang, Xin Zhao, Xin Dong, Hui Zhu, Yunnuan Shi, Hongyan Chen, Jianfei Shi, Da Feng, Li PLoS One Research Article The transmissible gastroenteritis virus (TGEV) nucleocapsid (N) protein plays important roles in the replication and translation of viral RNA. The present study provides the first description of two monoclonal antibodies (mAbs) (5E8 and 3D7) directed against the TGEV N protein linker region (LKR) and carboxyl terminal domain (CTD). The mAbs 5E8 and 3D7 reacted with native N protein in western blotting and immunofluorescence assay (IFA). Two linear epitopes, (189)SVEQAVLAALKKLG(202) and (246)VTRFYGARSSSA(257), located in the LKR and CTD of TGEV N protein, respectively, were identified after truncating the protein and applying a peptide scanning technique. Using mAb 5E8, we observed that the N protein was expressed in the cytoplasm during TGEV replication and that the protein could be immunoprecipitated from TGEV-infected PK-15 cells. The mAb 5E8 can be applied for different approaches to diagnosis of TGEV infection. In addition, the antibodies represent useful tools for investigating the antigenic properties of the N protein. Public Library of Science 2016-09-30 /pmc/articles/PMC5045181/ /pubmed/27689694 http://dx.doi.org/10.1371/journal.pone.0163920 Text en © 2016 Zhang et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Zhang, Xin
Zhao, Xin
Dong, Hui
Zhu, Yunnuan
Shi, Hongyan
Chen, Jianfei
Shi, Da
Feng, Li
Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title_full Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title_fullStr Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title_full_unstemmed Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title_short Characterization of Two Monoclonal Antibodies That Recognize Linker Region and Carboxyl Terminal Domain of Coronavirus Nucleocapsid Protein
title_sort characterization of two monoclonal antibodies that recognize linker region and carboxyl terminal domain of coronavirus nucleocapsid protein
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5045181/
https://www.ncbi.nlm.nih.gov/pubmed/27689694
http://dx.doi.org/10.1371/journal.pone.0163920
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