Preparation and identification of monoclonal antibody against pseudorabies virus UL36 protein
VP1/2,a tegument protein encoded by the pseudorabies virus UL36 gene,plays an important role in the assembly of virions.In this study,UL36 gene was mainly used as the research object,the truncated UL36 gene was cloned into pCold I vector to construct the recombination prokaryotic expression plasmid pCold I-UL36.After induced expression,the recombinant protein expressed in inclusion body form was denatured,affinity chromatography and renaturation to obtain high purity protein.Then,BALB/c mice were immunized with purified protein.A hybridoma cell strain named UL36-6,stably secreting antibodies against UL36 was obtained by cell fusion and indirect ELISA screening.The identification of antibody subtype showed that it was IgG1 and κ.The ascites antibody had high neutralization titer and purity.The Western-blot and indirect immunofluorescence test results showed that the monoclonal antibody had good specificity.In conclusion,we successfully prepared a monoclonal antibody against PRV UL36 protein in the present study which will lay a foundation for further study of the biological function of UL36 protein.
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