1.Generation and characterization of monoclonal antibody against glycoprotein VP1 encoded by caprine enterovirus CEV-JL14
Haibing LU ; Mingyue WANG ; Lisai ZHU ; Yajing LIU ; Changming GUO ; Qun ZHANG ; Yue YUAN ; Changchun TU ; Xinping WANG
Chinese Journal of Veterinary Science 2017;37(8):1468-1472
VP1 gene encoded by the newly identified caprine enterovirus CEV-JL 14 was amplified and cloned to prokaryotic expression vector pGEX-4T-1.Recombinant GST-VP1 fusion protein was expressed,purified,emulsified with Freund's complete adjuvant and used to immunize the BALB/c mice following a standard procedure.The spleen cells from immunized mice were collected and fused with myeloma cells after its antibody titer reached over 104 times detected by indirect ELISA.Hybridoma cell clones secreting monoclonal antibodies against VP1 were screened and their stability and specificity were further determined.The identified hybridoma cells were injected to mice intraperitoneally and ascites were collected at 7 DPI.Isotypes of the monoclonal antibodies against the recombinant VP1 protein were characterized to be either IgG1 or IgG2b,which showed a high specificity for detection of caprine enterovirus antigens by immunoperoxidase monolayer assay,thus laying a solid basis for future study related to viral pathogenesis,detection and diagnostics for caprine enterovirus infection.
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