[Preparation and application of anti-envelope protein B2L mouse polyclonal antibody of ORF virus].

: Objective To obtain the envelope protein B2L of orf virus(ORFV) and prepare highly specific polyclonal antibody against B2L. Methods The B2L gene was amplified by PCR, and subcloned into the vectors pET-32a and p3×FLAG-CMV-14, pET-32a-B2L followed by expression of B2L protein in E.coli, and induction of the target protein by IPTG, purification by urea solution and identification via Western blot analysis. Furthermore, the BALB/c mice were immunized with B2L protein to prepare highly-specific anti-B2L polyclonal antibody. In addition, plasmid p3×FLAG-B2L was transfected into Vero cells and BHK-21 cells which was used to confirm its specificity and antigenicity by indirect immunofluorescence assay(IFA). Results Western blotting demonstrated that the B2L protein had high-level specificity. The results of IFA indicated that the anti-B2L specific polyclonal antibody had specificity and reactivity. Then Immunohistochemistry showed that it could neutralize natural ORFV. Conclusion Highly specific and purified polyclonal antibody against B2L protein of the ORFV was successfully prepared.