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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 用培养的人胆管癌细胞系SK CHA 1免疫BALB/c小鼠后 ,获取免疫小鼠的脾细胞 ,与小鼠骨髓瘤细胞SP 2 / 0融合 ,ELISA方法及免疫荧光染色法筛选细胞融合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杂交瘤细胞株分泌的单克隆抗体具有选择性强 ,效价高的特点 ,冻存半年后复苏 ,仍能稳定分泌。经免疫组织化学及免疫荧光检测发现 ,该单克隆抗体与胆管癌组织呈阳性反应 ,而与肝癌组织 ,胰腺癌组织以及其他消化道癌组织呈阴性反应。且与正常胆管组织及正常肝组织亦呈阴性反应。结论 2F4E8是一株稳定的杂交瘤细胞株 ,所分泌的抗人胆管癌相关抗原单克隆抗体特异性强 ,效价高 ,在胆管癌的诊断、治疗及基础研究中有较好的应用前景。
Objective To establish a hybridoma cell line capable of secreting monoclonal antibodies against human cholangiocarcinoma associated with human cholangiocarcinoma tissue. Methods BALB / c mice were immunized with cultured human cholangiocarcinoma cell line SK CHA 1, and the splenocytes of immunized mice were obtained and fused with mouse myeloma cell SP2 / 0. ELISA and immunofluorescence staining were used to screen the cell fusion The resulting hybridoma cell line. Results Five hybridoma cell lines secreting monoclonal antibodies against human cholangiocarcinoma-associated antigen were screened by ELISA and immunofluorescent staining. The monoclonal antibodies secreted by 2F4E8 hybridoma cell lines were highly selective and their titer High characteristics, frozen after six months of recovery, still stable secretion. Immunohistochemistry and immunofluorescence showed that the monoclonal antibody reacted positively with cholangiocarcinoma, but negative with HCC, pancreatic cancer and other digestive tract cancers. And with normal bile duct tissue and normal liver tissue also showed a negative reaction. Conclusion 2F4E8 is a stable hybridoma cell line. The secreted monoclonal antibody against human cholangiocarcinoma-associated antigen has strong specificity and high potency. It has good application prospect in the diagnosis, treatment and basic research of cholangiocarcinoma.