抑制素α、βA、βB亚基单克隆抗体制备及其初步应用  

Preparation and identification of monoclonal antibodies against synthetic human inhibin α、βA、βB fragments and its preliminary application

作　　者：夏辉明[1] 高晓东[1] 张东升[1] 王宏卫[1] 黄新[1] 张义国[1] 宫亚欧[1] 鲁桂琛[2] 孙颖[2] 

机构地区：[1]河南医科大学病理生理教研室,河南郑州450052 [2]中国医学科学院药物研究所,北京100050

出　　处：《中国病理生理杂志》2000年第11期1174-1177,I001,共4页Chinese Journal of Pathophysiology

基　　金：卫生部科学研究基金资助!(No .96-1-0 65 );河南省自然科学基金资助! (No .9840 2 0 2 0 0 )

摘　　要：目的 :制备INHα、βA、βB亚基的McAbs ,并进行肿瘤免疫组化研究。 方法 :抗原为猪卵泡液INH提取物及人工合成的INHα、βΑ、βB亚基肽段 (6个 )。以上述抗原免疫BALB/c小鼠 ,经细胞融合、筛选、克隆化 ,获得杂交瘤细胞 14株 ,对其免疫学特性进行了分析。用纯化的McAbs进行了肿瘤免疫组化研究 (2 85例次 )。结果 :获得分别分泌抗INHα、βA、βΒ亚基的McAbs细胞株 14个 ,效价 1× 10 -4 - 1× 10 -6。免疫组化除生殖系统肿瘤阳性外 ,还发现乳腺癌的阳性率超过 70 %。结论 :成功制备了分泌抗INHα、βΑ、βB亚基肽的McAbs 。AIM: Preparation and identification of McAbs against synthetic human inhibin α、βA、 or βB fragments and its immunohistochomical application in cancers.METHODS: The fragment of inhibin subunits α(1-26)(37-65), βA(1-28)(82-114),βΒ(1-28)(85-115) have been synthesized. Inhibin isolated from porcine follicle fluid and synthetic polypeptide fragments were used as antigen. Intraperitoneal immunization was performed on BALB/c mice, then boosted with intraspleen injection was performed 3 days prior to fusion. After fusion, selection and cloning procedures, 14 hybridoma cell lines were successfully obtained and their immunological characteristics were analyzed. After purification, McAbs were applied to immunohistochemical staining in gonadal tumors and other cancers.RESULTS: We obstained 14 stable cell lines which can secret McAbs againsting INH α、βA or βB subunit. The titre of ascites were 1×10 -4-1×10 -6. Immunohistochemical positive staining were observed in ovarian granulosa cell tumor (100%), ovarian mucinos tumor, embryonal carcinoma of the ovary, testis spermocytoma, embryonal carcinoma of the testis, In 40 samples of breast cancers, each INH subunit positive rates were over 70%.CONCLUSION: The INH's McAbs prepared by ourselves were usefull for cancer immunohistochemistry.

关 键 词：抑制素 单克隆抗体 肿瘤 免疫组化 亚基 McAbs 

分 类 号：R730.2[医药卫生—肿瘤] R392-33[医药卫生—临床医学]