Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid
- 15 March 2019
- journal article
- research article
- Published by Springer Nature in Applied Microbiology and Biotechnology
- Vol. 103 (9) , 3705-3714
Abstract
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.Keywords
Funding Information
- National Key R&D Program of China (2016YFD0500103)
- National Natural Science Foundation of China (31572498, 31702209)
- Elite Youth Program of CAAS (No)
This publication has 31 references indexed in Scilit:
- Origin, Evolution, and Genotyping of Emergent Porcine Epidemic Diarrhea Virus Strains in the United StatesmBio, 2013
- Emergence of Porcine epidemic diarrhea virus in the United States: clinical signs, lesions, and viral genomic sequencesJournal of Veterinary Diagnostic Investigation, 2013
- Sequence and phylogenetic analysis of nucleocapsid genes of porcine epidemic diarrhea virus (PEDV) strains in ChinaArchives of Virology, 2013
- New Variants of Porcine Epidemic Diarrhea Virus, China, 2011Emerging Infectious Diseases, 2012
- Outbreak of Porcine Epidemic Diarrhea in Suckling Piglets, ChinaEmerging Infectious Diseases, 2012
- Subcellular localization of the severe acute respiratory syndrome coronavirus nucleocapsid proteinJournal of General Virology, 2005
- Detection of the nucleocapsid protein of severe acute respiratory syndrome coronavirus in serum: Comparison with results of other viral markersJournal of Virological Methods, 2005
- The Coronavirus Infectious Bronchitis Virus Nucleoprotein Localizes to the NucleolusJournal of Virology, 2001
- Coronavirus immunogensVeterinary Microbiology, 1993
- Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assayJournal of Immunological Methods, 1985