抗乳腺癌单克隆抗体的制备及其应用

Preparation and Applicationof Monoclonal Antibody Against Human Breast Cancer in Tumor Research

目的 :制备抗人乳腺癌单克隆抗体(McAb) ,用于临床肿瘤的免疫病理诊断与分型研究 ,为药物导向诊断和治疗奠定基础。方法 :以乳腺癌细胞系ZR75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、筛选及克隆化 ,获得恒定分泌抗ZR75单克隆抗体的杂交瘤细胞系BM109。结果 :单抗BM109的Ig亚类为IgG1,BM109抗原主要表达在靶细胞膜上 ,为分子量37KD的蛋白质抗原。在乳腺癌的免疫反应率为86.7 %(26/30) ,与胃癌、食管癌、卵巢癌等6种肿瘤的免疫反应率为9.5 %(4/42) ,而对乳腺纤维瘤、囊性增生症和周围正常乳腺组织未见阳性反应。结论 :所获一株抗乳腺癌单克隆抗体 ,具有较好的特异性 ,可望进一步用于乳腺癌的诊断。

Objective:To prepare monoclonal antibody (McAb) of human breast cancer associated antigen and to lay a foundation for further study of drug guided diagnosis and treatment.Methods:Balb/c mice were immunized with human breast cancer cell line ZR75 and the lymphocytes hybridoma technique was used for McAb preparation.The immunofluorescence immunohistochemistry (IHC) and Western blotting methods were used for the characterization of antigen recognized by McAb BM109.Results:One hybridoma cell line (BM109),which secretes monoclonal antibody against breast carcinoma,was established.McAb BM109 was found to be the IgG1 isotope by double diffusion.The antigen recognized by BM109 was a protein with a molecular weight of 37KD and located on ZR75 cell membrane.The reactivity of BM109 to breast cancer and several other kinds of tumor were examined with IHC,the results showed that the BM109 reacted strongly to most breast carcinoma and did not react to normal breast tissues and other kinds of tumor,such as gastric cancer、esophageal cancer、lung cancer and other three tumors.Conclusion:The hybridoma cell line BM109 secreting monoclonal antibody stably was obtained,The McAb BM109 could react specifically with breast carcinoma and might be used in basic and clinical researches of breast cancer with a bright prospect.