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

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.