Preparation of functional antibodies targeting tumor endothelia for the treatment of hepatic carcinoma

BACKGROUND & OBJECTIVE: Tumor growth relies on angiogenesis; angiogenic inhibition can effectively treat tumors. This study was to develop monoclonal antibody library against human hepatic carcinoma vessel endothelial cells (HCVECs), and screen functional monoclonal antibodies with inhibitory effect on the angiogenesis of hepatic carcinoma. METHODS: Endothelial cells were separated from human hepatic carcinoma tissue to immunize 10 BALB/c mice. The spleen cells of the immunized mice were fused with SP2/0 cells and cultured on methyl cellulose. Functional monoclonal antibodies of each clone were screened by immunofluorescence, endothelial cell proliferation assay, tube formation assay, and humanized blood vessel in vivo tumor model. RESULTS: A total of 1,442 monoclonal antibody clones were obtained by fusing spleen cells with SP2/0 cells; 119 clones could specifically react with HCVECs. Of the 119 clones, 53 significantly suppressed the proliferation or tube formation of HCVECs. Two of the 53 monoclonal antibody clones suppressed the hepatic tumor growth in vivo, with inhibitory rates of 66.7% and 76.5%. CONCLUSION: We successfully developed 2 functional monoclonal antibodies which could suppress the hepatic tumor growth in vivo.