Study on ScFv of anti bladder cancer J82 monoclonal anti body and anti mitomycin C monoclonal antibody

Objective To construct the single chain f ragment of variable region, which showed a much lower antigenictiy and may be a practical solution for target therapy of malignant tumor.Methods Hybridoma were generated by fusing the immunized Balb/c mice splenocytes and myeloma. The genes of variable chains were cloned with PCR. And the ScFv ge ne of the antibodies were constructed by using PCR-SOE(splicing by overlap ext ension).Results The established cell lines of D10 1C8 could secret anti J82 monoclonal antibody and anti Mitomycin C (MMC) monoclonal antibody. The activity and specificity of the antibodies were confirmed by ELIS A and histochemistry method. The genes of the variable chain (320 bp and 340 bp fragments)and the ScFv (750 bp fragment) of the two antibod ies were obtained.Conclusion The ScFv gene of the antibod ies against bladder cancer and MMC could be obtained conveniently and quickly by using PCR-SOE method, which may be helpful to the research on bi-specific ScF v anti bladder cancer-MMC.