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目的 研制抗c erbB 2胞内区多肽单克隆抗体 ,用于乳腺癌等c erbB 2过量表达的诊断及指导治疗。方法 合成p185蛋白胞内区多肽作为抗原 ,免疫BALB/C小鼠建立了一组分泌抗该多肽的特异性高亲和力单克隆抗体杂交瘤细胞系 ,并对该单抗的生化特性及其免疫学反应性进行了研究和测定 ,并与美国食品药品管理局 (FDA)批准的抗c erbB 2多克隆抗体进行了比较。结果 共获得3株稳定分泌抗p185胞内区单抗的杂交瘤细胞株。其中 0 35 E6 1株单抗经对 39例 (T1 2 N0 M0 )乳腺癌患者组织病理切片进行免疫组织化学染色 ,c erbB 2过量表达阳性率为 2 6 % ,而乳腺纤维腺瘤组织切片没有非特异染色 ;流产正常胎儿器官组织切片除甲状腺、肾上腺和肾小管上皮表达外 ,其余均不表达。与FDA批准的DAKO公司多抗的阳性染色符合率为 74%。结论 0 35 E6 1单抗可特异识别乳腺癌细胞内p185胞内区抗原。对判断乳腺癌预后及指导治疗可能具有应用价值。
OBJECTIVE: To develop a monoclonal antibody against c erbB 2 intracellular domain for the diagnosis and treatment of overexpression of c erbB 2 in breast cancer. Methods The intracellular domain of p185 protein was synthesized and used as antigen to immunize BALB / C mice to establish a group of hybridoma cell lines secreting the specific high affinity monoclonal antibody against the polypeptide. The biochemical characteristics and immunology Reactivity was studied and assayed and compared to the anti-c erbB 2 polyclonal antibody approved by the U.S. Food and Drug Administration (FDA). Results Three hybridoma cell lines stably secreting anti-p185 intracellular mAb were obtained. The 0 35 E6 monoclonal antibody was immunohistochemically stained by immunohistochemistry in 39 cases (T1 2 N0 M0) of breast cancer patients, the positive rate of c erbB 2 overexpression was 26%, while that of breast fibroadenoma did not Non-specific staining; abortion normal fetal organ tissue sections in addition to the thyroid, adrenal and renal tubular epithelial expression, the rest were not expressed. In line with the FDA approved DAKO polyclonal positive staining in line with the rate of 74%. Conclusion 0 35 E6 1 McAb can specifically recognize intracellular p185 antigen in breast cancer cells. To determine the prognosis of breast cancer and guiding treatment may have value.