Abstract: Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen Glypican-3 (GPC3) and preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. coliBL21. Spleen cells were obtained from Balb/c mice embedding immunized with purified antigen intraperitoneally, and fused with SP2/0 cells. Hybridoma cells were screened by indirect ELISA, and identified by Western blot using purified protein after the cell fusion. Indirect immunofluorescence method was used to test the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stablely expressed in E. coliBL21. A mice hybridoma cell line producing monoclonal antibody to GPC3 was obtained. Western blot analysis showed that monoclonal antibody specifically recognized recombinant protein. Monoclonal antibody could be used to detect GPC3 protein expression in HepG2 cell line by indirect immunofluorescence. Conclusion Monoclonal antibody against GPC3 was obtained. The research will be useful for its clinical study.

Key words: Glypican-3, Embedding immunology, Monoclonal antibody