一株抗人胆管癌相关抗原杂交瘤细胞株的建立及鉴定

目的 建立能分泌与人胆管癌组织特异结合的高效价抗人胆管癌相关抗原单克隆抗体的杂交瘤细胞株。方法 用２的人胆管癌细胞系Ｓｋ－ＣＨＡ－１免疫ＢＡＬＢ／ｃ小鼠后，获取免疫小鼠的脾细胞。与小鼠骨髓瘤ＳＰ２／０融合，ＥＬＩＳＡ方法及免疫荧光染色法筛选细胞晤后生成的杂交瘤细胞株。结果 ＥＬＩＳＡ方法及免疫荧光染色法共筛选出５株能持续稳定分泌抗人胆管癌相关抗原单克隆抗体的杂交瘤细胞株，其中２Ｆ４Ｅ８杂交瘤细胞株

Establishment and identification of a hybridoma cell line against human cholangiocarcinoma-related antigen

Objective To produce an anti cholangiocarcinoma related antigen hybridoma cell line which can secreate antibody specifically bind to human bile duct carcinoma tissues with high titer. Methods Spleen cells were obtained from BALB/c mice immunized with human cholangiocarcinoma cell line SK CHA 1 and fused with murine myeloma cell SP 2/0. Hybridoma cells were selected with ELISA and immunofluoresent staining after the cell fusion. Results Five clonies of hybridoma cell which could secrete anti cholangiocarcinoma related antigen McAb were obtained. Among these colonies, the hybridoma cell line 2F4E8 was found to secrete McAb with the same specificity and titer after 6 months of freezing. With immunohistochemical and immunofluoresent assay, it was found that this McAb could positively react with bile duct carcinoma tissues, while negatively with hepatocarcinoma, pancreatocarcinoma and other gastroenteric malignant tissues, and also has negative reaction with normal bile duct and liver tissues. Conclusion 2F4E8 is a stable hybridoma cell line and the anti cholangiocarcinoma related antigen McAb it secretes is of high specificity and titer. This cell line possesses the prospect in the diagnosis, treatment and basic research of cholangiocarcinoma.