Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Wenting, Chen, Wenwen, Huang, Jingling, Jin, Li, Zhou, Yawei, Chen, Jianing, Zhang, Na, Wu, Donglai, Sun, Encheng, Liu, Guangliang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2019
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
_version_ 1783507920646832128
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
work_keys_str_mv AT yangwenting generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT chenwenwen generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT huangjingling generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT jinli generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT zhouyawei generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT chenjianing generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT zhangna generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT wudonglai generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT sunencheng generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid
AT liuguangliang generationidentificationandfunctionalanalysisofmonoclonalantibodiesagainstporcineepidemicdiarrheavirusnucleocapsid