Anti-Estrone-sulfalase monalonal antibody

子宫内膜癌组织在高雌激素-硫酸酯酶活性的基础上显示局部高雌激素活性。然而，由于抗甾酮-硫酸酯酶单克隆抗体尚不存在，因此该酶的组织学定位尚不清楚。本研究的目的是制备单克隆抗体，以证明在子宫内膜癌组织中具有高活性的雌激素-硫酸酯酶。子宫内膜炎中雌酮硫酸酯酶的组织学定位。10个胎盘均质，用柱层析法结块。硫酸h3雌酮酶分析显示雌酮硫酸酯酶富峰较高。洗脱峰上的蛋白，作为生产单克隆抗体的抗原。用该蛋白免疫BALB/c nu/nu小鼠，采用ELISA法和western-blotting筛选12个克隆细胞。免疫组化结果显示，所有单克隆抗体对正常胎盘有反应，对先天性鱼鳞病胎盘无反应。用芳基硫酸酯酶c基因转染乳腺癌细胞株（MCF-7和T47D），结果表明，与单独转染载体相比，芳基硫酸酯酶c基因对雌激素-硫酸酯和脱氢表雄酮的活性更高。单克隆抗体与来自转染物的酶蛋白发生反应。这证明单克隆抗体同时检测到雌激素-硫酸酯酶和脱氢表雄酮，子宫内膜组织和肿瘤的免疫组化染色显示正常子宫内膜组织无活性，但子宫内膜癌组织，特别是单独的癌细胞对抗体有反应。提示雌激素-硫酸酯酶定位于子宫内膜癌上皮细胞。综上所述，我们成功地制备了抗雌酮硫酸酯酶单克隆抗体。

Endometrial carcinoma tissues showed locally high activity of estrogen based on high activity of estrtone-sulfatase. However, histrological localization of the enzyme has not been unknown, because anti-cstrone-sulfatase monoclonal antibody is not available yet. The purpose of present study is the production of the monoclonal antibody and in order to demonstrate the High-activity of estrone-sulfatase in endometrial carcinoma tissues has been reported.histological localization of estrone sulfatase in endomentrial tidsses. Ten placentas were homogenized and cluted by columchromatography. Enzyme assay with H3-estrone-sulfate showed a high rich peak of estrone-sulfatase. The protein on the peak were eluted and was used as an antigen for the production of monoclonal antibody. BALB/c nu/nu mice were immunized with the protein and then twelve clone cells were selected by both ELISA and western-blotting. Immunohistochemistry showed that the all monoclonal antibodies were reactive to normal placenta, but inactive to placentas of congenital ichtyosis. Transfection with aryl-sufatase-C gene to breast carcinoma cell lines (MCF-7 and T47D) showed higher activities of both estrone-sulfatse and dehydroepiandrosterone than transfectants with vector alone. The monoclonal antibodies reacted on the enzyme proteins from the transfectants. This proves that the monoclonal antibodies were detected both estrone-sulfatase and dehydroepiandrosterone, Immunohistochemical staining of endometrial tissues and neoplasms demonstrated that normal endometrial tissues were inactive, but endometrial acrcinoma tissues, especially carcinoma cells alone, were reactive to the antibody. This suggests that estrone-sulfatase are localized on epithelial cells of endometrial carcinomas. In conclusion, we succeeded in the production of anti-estrone-sulfatase monoclonal antibodies.