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Preparation and identification of a 9B9 monoclonal antibody specifically targeting EGFRvⅢ/EGFR
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Abstract:
Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor (EGFR) and (or) epidermal growth factor receptor vⅢ(EGFRvⅢ), and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7EGFRvⅢ cell and epidermal carcinoma A431 cellimplanted tumors in nude mice. Methods:BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ (3T3EGFRvⅢ). Immunized spleen cells were fused with myeloma SP2/0 cells, and antiEGFRvⅢ monoclonal antibody positive hybridoma cells (named 9B9 antibody and 9B9 cells, respectively) were selected and identified by ELISA. The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay. Huh7EGFRvⅢ cell(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cellbearing mouse models were established and were divided into PBS group, Cetuximab group and 9B9 antibody group. Then, antitumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab. Results:A monoclonal antibody, named 9B9 antibody, was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence. The inhibitory rates of Cetuximab and 9B9 antibody against Huh7EGFRvⅢ cellsimplanted tumors were 42% and 46%, respectively, and those against A431 cellsimplanted tumors were 85% and 86%, respectively. Conclusion:9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell and epidermal cell carcinoma cellimplanted tumors, and the effects resemble that of Cetuximab.
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Project Supported:
Supported by the Shanghai Rising Star Foundation for Young Scholar (type A) (No. 07QA14046); the National High Technology Research and Development Program (863 Program) of China (No.2007AA021203); the Science Foundation of Shanghai Scientific Committee (N
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