摘要
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 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.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2000年第7期619-622,共4页
Journal of Third Military Medical University
基金
国家自然科学基金!资助项目 (39670 71 3)
