Anti-Estrone-sulfalase monalonal antibody

子宫内膜癌组织中雌激素活性高，雌激素酯硫酸酯酶活性高。然而，由于抗雌酮硫酸酯酶的单克隆抗体尚未获得，因此该酶的组织学定位并不是未知的。本文报道了雌酮硫酸酯酶在子宫内膜癌组织中的组织学定位，并以雌酮硫酸酯酶单克隆抗体的制备为目的，证实了雌酮硫酸酯酶在子宫内膜癌组织中的高活性。取10例胎盘匀浆，柱层析分离。用H3-雌酮硫酸酯酶测定显示雌酮硫酸酯酶的高富集峰。洗脱峰上的蛋白质，并将其用作单克隆抗体生产的抗原。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。免疫组化结果显示，所有单克隆抗体对正常胎盘均呈阳性反应，对先天性鱼鳞病胎盘均呈阴性反应。将芳基硫酸酯酶C基因转染乳腺癌细胞系MCF-7和T47 D，其雌酮硫酸酯酶和脱氢表雄酮的活性均高于单独转染载体的细胞。单克隆抗体与来自转染子的酶蛋白反应。子宫内膜组织和子宫内膜癌组织的免疫组化染色显示，正常子宫内膜组织无活性，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。总之，我们成功地生产抗雌酮硫酸酯酶的单克隆抗体。

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.