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Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid
The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7079923/ https://www.ncbi.nlm.nih.gov/pubmed/30877355 http://dx.doi.org/10.1007/s00253-019-09702-5 |
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author | Yang, Wenting Chen, Wenwen Huang, Jingling Jin, Li Zhou, Yawei Chen, Jianing Zhang, Na Wu, Donglai Sun, Encheng Liu, Guangliang |
author_facet | Yang, Wenting Chen, Wenwen Huang, Jingling Jin, Li Zhou, Yawei Chen, Jianing Zhang, Na Wu, Donglai Sun, Encheng Liu, Guangliang |
author_sort | Yang, Wenting |
collection | PubMed |
description | The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies (mAbs) targeting N protein of PEDV and analyzed their applications on enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), western blot assay, and flow cytometry assay. The results demonstrated that all these six mAbs were IgG1 isotype and κ chain. Among these six mAbs, 3F12 recognizes a linear epitope (VAAVKDALKSLGI) while the other five mAbs recognize different conformational epitopes formed by a specific peptide fragment or the full length of N protein. The functional analysis showed that all these six mAbs were applicable to ELISA, western blot, IFA, and flow cytometry assay. In conclusion, we developed six mAbs against PEDV-N protein to facilitate the early detection of PEDV infection using ELISA, western blot, IFA, and flow cytometry. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09702-5) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7079923 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-70799232020-03-23 Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid Yang, Wenting Chen, Wenwen Huang, Jingling Jin, Li Zhou, Yawei Chen, Jianing Zhang, Na Wu, Donglai Sun, Encheng Liu, Guangliang Appl Microbiol Biotechnol Biotechnological Products and Process Engineering The variant strains of porcine epidemic diarrhea virus (PEDV) severely threaten the pig industry worldwide and cause up to 100% mortality in suckling piglets. It is critically important and urgent to develop tools for detection of PEDV infection. In this study, we developed six monoclonal antibodies (mAbs) targeting N protein of PEDV and analyzed their applications on enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), western blot assay, and flow cytometry assay. The results demonstrated that all these six mAbs were IgG1 isotype and κ chain. Among these six mAbs, 3F12 recognizes a linear epitope (VAAVKDALKSLGI) while the other five mAbs recognize different conformational epitopes formed by a specific peptide fragment or the full length of N protein. The functional analysis showed that all these six mAbs were applicable to ELISA, western blot, IFA, and flow cytometry assay. In conclusion, we developed six mAbs against PEDV-N protein to facilitate the early detection of PEDV infection using ELISA, western blot, IFA, and flow cytometry. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00253-019-09702-5) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2019-03-15 2019 /pmc/articles/PMC7079923/ /pubmed/30877355 http://dx.doi.org/10.1007/s00253-019-09702-5 Text en © Springer-Verlag GmbH Germany, part of Springer Nature 2019 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 | Biotechnological Products and Process Engineering Yang, Wenting Chen, Wenwen Huang, Jingling Jin, Li Zhou, Yawei Chen, Jianing Zhang, Na Wu, Donglai Sun, Encheng Liu, Guangliang Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title | Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title_full | Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title_fullStr | Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title_full_unstemmed | Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title_short | Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
title_sort | generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid |
topic | Biotechnological Products and Process Engineering |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7079923/ https://www.ncbi.nlm.nih.gov/pubmed/30877355 http://dx.doi.org/10.1007/s00253-019-09702-5 |
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