Preparation and Preliminary Application of Monoclonal Antibody Against Carcinoembryonic Antigen Glypican-3 / 天津医药

ABSTRACT

Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen glypican-3 (GPC3) and its preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. co-liBL21. 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 assay using puri-fied protein after the cell fusion. The indirect immunofluorescence method was used to detect the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stably expressed in E. coliBL21. A mouse hybridoma cell line secreting 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 The monoclonal antibody against GPC3 was successfully obtained.