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Preparation and Identitficaion of Monoclonal Antibody against VP7 Protein of Group A Porcine Rotavirus

CHI Yan-bin1,2,CHEN Jian-fei2,SHI Hong-yan2,ZHANG Xin2,SHI Da2,LI Chang-long2,HAN Xiao2,CHEN Hong-yan1,2,FENG Li1,2   

  1. 1.College of Life Science,Northeast Agricultural University,Harbin 150030,China;2.Division of Swine Infectious Diseases,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China)
  • Received:2013-06-26 Online:2014-02-20 Published:2014-03-27

Abstract: To prepare the monoclonal antibodies against VP7 protein of group A porcine rotavirus (PoRV) serotype G11,VP7 gene of PoRV serotype G11 was cloned into the vector pGEX-6P-1. The recombinant plasmid was transformed into E.coli competent cells DH5α and induced by IPTG. Hybridomas were produced by fusing SP2/0 cells with spleen cells from mouse immunized with GST-VP7 recombinant protein. One hybridomas secreting MAb against VP7 protein was identified by indirect ELISA. Western blotting analysis showed that the MAb could recognize the recombinant VP7 protein and authentic VP7 protein of PoRV,and the specific immunoflurescence was detected in PoRV infected MA104 cells by indirect immunoflurescence assay.The result of MTT method showed that the MAb had the neutralizing activity. The subtype of the MAb was IgG2b. The results showed that VP7 protein was successfully expressed in E.coli, one MAb was preparated and identified,and it could be used for further study of prevention,diagnosis and treatment of porcine rotavirus disease.

Key words: group A procine rotavirus; VP7 gene; monoclonal antibodies