Anti-Estrone-sulfalase monalonal antibody

子宫内膜癌组织表现出局部高雌激素活性，其基础是雌酚硫酸酯酶高活性。然而，该酶的历史定位尚不清楚，因为目前还没有抗cstrone-硫酸酯酶的单抗。本研究的目的是制备单抗，为了证明雌酮-硫酸酯酶在子宫内膜癌组织中的高活性，报道了雌酮-硫酸酯酶在子宫内膜滑液中的组织定位。用柱层析法对10个胎盘进行匀浆和线索分析。H3-雌酮-硫酸盐酶活测定结果表明，雌酮-硫酸酯酶具有较高的富集峰。将峰上的蛋白洗脱后作为抗原制备单抗。用该蛋白免疫BALB/c nu/nu小鼠，经ELISA和Western-blotting筛选出12个克隆细胞。免疫组织化学结果显示，所有单抗对正常胎盘均呈阳性反应，而对先天性硬膜外胎盘均无反应。转导芳基硫酸酯酶C基因的乳腺癌细胞株MCF-7和T47D的硫酸雌酮和脱氢表雄酮活性均高于单纯载体转染组。单抗可与转染体中的酶蛋白发生反应。免疫组织化学染色显示正常子宫内膜组织不活跃，而子宫内膜癌组织，尤其是癌细胞对该抗体呈阳性反应。这表明雌酮-硫酸酯酶定位于子宫内膜癌的上皮细胞。综上所述，我们成功地制备了抗雌酮-硫酸酯酶的单抗。

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.