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以食管癌“109”细胞株免疫 BALB/C 小鼠,取其脾细胞与小鼠骨髓瘤细胞株SP2/0 融合,经三次克隆化,获得稳定分泌抗食管癌“109”细胞株单克隆抗体的杂交瘤细胞系2GF3。其分泌的单克隆2GF3抗体与外周血淋巴组胞、ABO 型红细胞、癌胚抗原以及3/5胃癌细胞株无反应,与2/5胃癌细胞株有弱(+)反应。与2GF3抗体对应的抗原为2GF3抗原,荧光标记检测表明后者主要表达在细胞膜上。用去污剂提取膜抗原进行电泳后转移至硝酸纤维素膜上进行酶标分析,2GF3抗原是分子量53KD 的糖蛋白,在还原及非还原条件下均显示相同的区带。它与抗癌胚抗原抗体及抗甲胎蛋白抗体均无反应,表明它不同于癌胚抗原和甲胎蛋白。
BALB / C mice were immunized with esophageal carcinoma cell line “109”. The spleen cells were fused with mouse myeloma cell line SP2 / 0 and cloned three times to obtain stable monoclonal antibody against esophageal cancer cell line “109” Of the hybridoma cell line 2GF3. The secreted monoclonal 2GF3 antibody did not react with peripheral blood lymphocytes, ABO erythrocytes, carcinoembryonic antigen and 3/5 gastric cancer cell lines, but weakly (+) reacted with 2/5 gastric cancer cell lines. The antigen corresponding to the 2GF3 antibody was 2GF3 antigen, and the fluorescent labeling assay showed that the latter was mainly expressed on the cell membrane. The membrane antigen was extracted with detergent and then transferred to nitrocellulose membrane for enzyme labeling analysis. The 2GF3 antigen was a 53KD glycoprotein with the same band under both reducing and non-reducing conditions. It is non-responsive to both the anti-CEA antibody and anti-alpha-fetoprotein antibody, indicating that it is distinct from carcinoembryonic antigen and alpha-fetoprotein.