Generation, identification, and functional analysis of monoclonal antibodies against porcine epidemic diarrhea virus nucleocapsid
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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.
KeywordsPorcine epidemic diarrhea virus Monoclonal antibodies Nucleocapsid protein
This work was supported by the National Key R&D Program of China (2016YFD0500103), the National Natural Science Foundation of China (31572498, 31702209), and the Elite Youth Program of CAAS.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
All the mouse experimental procedures were approved by the Lanzhou Veterinary Research Institute experimental animal committee. All applicable international, national, and institutional guidelines for the care and use of animals were strictly followed.
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