Preparation of monoclonal antibodies against VP2 protein of Asia I type foot-and-mouth disease virus and establishment of a competitive ELISA for the detection of antibodies / 生物工程学报

ABSTRACT

Monoclonal antibodies against FMDV vp2 protein were prepared and a competitive ELISA based on the monoclonal antibodies and vp2 protein was established. Balb/c mice were immunized with Escherichia coli expressed fusion protein. The splenocytes from immunized mice were fused with myeloma cells SP2/0. The hybridism cells were screened by indirect ELISA and limited dilution method. Two hybndoma cell Iines secreting mAbs against Asia I type foot-and-mouth disease were obtained. The titer and relative affinity of mAbs were determined by ELISA. Specificity of mAbs was analyzed by Western blotting. The ELISA titers of the ascites induced by the two hybridism cells were above 100 x 2(9).A competitive ELISA for the use of FMDV antibody detection was established using E. coli expressed fusion protein as coating antigen and HRP-labled mAb as detecting antibody. Clinical tests showed the method had 89.0 percent agreement with UBI Kit to detection of FMDV antibodies and 86.5 percent agreement with LPB- ELISA kit (Ceditest kit) for detection of antibodies against Foot-and-Mouth Disease Virus respectively.