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以无血清培基培养胃癌细胞M85,收集脱落抗原。以胃癌细胞及其脱落抗原免疫Balb/C小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2/0进行融合。经选择培养后,以免疫荧光染色及脱落抗原固相ELISA进行双重筛选。三次克隆化后获得稳定地分泌单克隆抗体(MCAb)的杂交瘤系S_1、S_2及S_3。McAb S_1、S_2及S_3主要与胃癌细胞系呈阳性反应而不与正常细胞及经检测的其它肿瘤细胞起反应。其对应抗原定位于靶细胞的膜上且可释放于培养上清中。
Gastric cancer cell M85 was cultured with serum-free medium and shedding antigen was collected. Balb/C mice were immunized with gastric cancer cells and their shedding antigens, and their spleen cells were fused with mouse myeloma cells SP2/0. After selective culture, double screening was performed by immunofluorescence staining and shed antigen solid phase ELISA. After three times of cloning, hybridomas S_1, S_2, and S_3 stably secreting monoclonal antibodies (MCAbs) were obtained. McAb S_1, S_2 and S_3 are mainly positive with gastric cancer cell lines but do not react with normal cells and other tumor cells detected. The corresponding antigen is located on the membrane of the target cell and can be released in the culture supernatant.