The Preparation of Monoclonal Antibody against ORF2-V1 Recombinant Proteins of Swine Hepatitis E Virus and Differential Recognition of Antigenic Epitopes

The BALB/c mice were immuned with purified recombinant fusion protein (ORF2-V1)from swine Hepatitis E virus. The myeloma cell line SP2/0 was fused with the spleen of the immuned BALB/c mice, and the positive clones which produced McAbs against ORF2-V1 were screened by ELISA. The titers of the six McAbs in the supernatant culture supernatant were 1∶1.60×103～1∶3.20×103, respectively , and the titers in the ascites of the mice were 1∶1.28×106～1∶2.56×106, respectively, detected by ELISA. And six McAbs only could react with ORF2-V1, and but not react with other recombinant fusion protein from ORF2. Among them , The A549 cell culture affected with HEV was carried out comparative studies, which is detection of pathogene with indirect immunofluorescence by using monoclonal antibody against HEV ORF2-V1 protein and polyclonal antiserum against HEV. The result shows that McAb against HEV ORF2-V1 protein, its speciality is high. Six monoclonal antibody belonged to IgG1, The titers of the six McAbs in the supernatant culture supernatant were 1∶1.60×103～1∶3.20×103, and the titers in the ascites of the mice were 1∶1.28×106～1∶2.56×106. The six strains of McAbs were specific for three different kinds of epitopes on HEV. Among them, McAb-γH1, McAb-BC4 and McAb-CH8 direct to different epitopes, respectively.The epitope recognized by McAb-γH1 overlapped partly that of McAb-BC4.McAb-αC11 and McAb-αC12 were identical to McAb b-γF8.The epitope of which was supposed to be locateed in the overlapping region between epitope-γH1 and epitope-BC4.