Preparation and Characterization of Monoclonal Antibodies Against Porcine Epidemic Diarrhea Virus

This study aimed to obtain the monoclonal antibodies against porcine epidemic diarrhea virus(PEDV) for antibody characterization as well asdisease control.The antigen of PEDV was purifiedby means of differential centrifugation and sucrose gradientcentrifugation. Spleen cells of the Balb/c mice immunized with viral antigen were fused with SP2/0 myeloma cells.Hybridoma cell lines secreting the monoclonal antibodies were isolated by using the indirect ELISA and sub-cloning approach to provide the source for the target antibody.Subsequently, specificities of the antibody were determined by means of IFA, ELISA and the Western blotting. The results obtained the following. (a) Two isolated hybridoma cell lines that could steadily secretethe specific McAbs against PEDV were designated as E1 and H6. (b) The isotyping analysis showed that E1 belonged to IgG2a; and H6, IgM sub-type. The McAbs from them could both bind to the antigen of PED in ELISA, IFA and the western blot analyses. (c) ELISA titers of the supernatants of E1 and H6 were 2⁶ and 2⁴ respectively; while those of the ascites fluids of E1 and H6, 10⁵ and 10⁴ respectively. (d) The McAbs from E1 and H6 showed no cross-reactivity with Vero cells, CSFV, TGEV or PRRSV.(e) The Western blotting confirmed that E1 McAbs recognized M protein; and, H6 McAbs, N protein. It appeared that two monoclonal antibodies with a high specificity against PEDV were successfully prepared, and further studies on PEDV immunodiagnosis, epitope discernment and the proteins became possible.