抗乳腺癌单抗的制备及应用

Preparation of Monoclonal Antibody Against Human Breast Cancer and its Application 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 ,采用淋巴细胞杂交瘤技术制备单克隆抗体。用免疫荧光、免疫组化和免疫印迹法研究其识别抗原特性。结果 :获得恒定分泌抗ZR75的杂交瘤细胞系BM 1 0 9。单抗BM 1 0 9属IgG1类型 ,其识别抗原是分子量为 37KD的蛋白性抗原 ,主要表达在靶细胞膜上。在乳腺癌的免疫反应率为 86 7% (2 6 / 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 :获得 1株较特异的抗乳腺癌单克隆抗体 ,该McAb对乳腺癌特异性高 ,对其他肿瘤交叉反应较小 ,可望进一步用于乳癌的诊断治疗以及发生发展的研究。

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:Immunization of Balb/c mice with of human breast cancer cell line ZR75,the study applied lymphocytes hybridoma techniques in McAb preparation and used immunofluorescence、immunohistochemistry (IHC) and Western blotting in 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 of the IgG 1 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 tumors was 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 tumors,such as gastric cancer、esophagus cancer、lung cancer and other three tumors.Conclusion:The hybridoma cell line BM109 stably secreting antibody reacted specifically with breast carcinoma was obtained and the McAb BM109 might be used in basic and clinical researches of breast cancer with a bright prospect. [