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本研究用乙醇法从人脑垂体丙酮干粉中提取粗品。用CM-52纤维素离子交换层析法分离促卵泡激素(FSH)和促黄体激素(LH),再用SP-Sephadex-C_(50)离子交换葡聚糖凝胶层析法纯化FSH,用DEAE-52纤维察离子交换层析法纯化LH,用HPLC蛋白A亲和层析法纯化抗LH和抗FSH免疫球蛋白,最后用HPLC抗LH的IgG和抗FSH的IgG精制LH和FSH。精制的FSH放免活性为5406Iu/mg(国际标准品4292),交叉反应(W/W)与LH为1.2％(国际标准品<3.1％)与TSH为0.46％(国际标准品>0.7％)。精制的LH放免活性为7928Iu/mg(国际标准品6640),交叉反应(W/W)与FSH<0.5％(国际标准品<2％)与TSH<0.1％(国际标准品<1％)。本研究制备的人脑垂体FSH和LH纯品达到了国际上同类产品水平,填补了国内空白。
In this study, crude ethanol was extracted from human pituitary acetone powder. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were separated by CM-52 cellulose ion-exchange chromatography and purified by SP-Sephadex-C 50 ion exchange gel chromatography LH was purified by ion exchange chromatography on DEAE-52 fibers, anti-LH and anti-FSH immunoglobulins were purified by HPLC Protein A affinity chromatography and finally LH and FSH were purified using HPLC anti-LH and anti-FSH IgG. The FSH activity of the purified FSH was 5406Iu / mg (International Standard 4292). The cross-reaction (W / W) and LH were 1.2% (international standard <3.1%) and TSH 0.46% (international standard> 0.7%). Purified LH radioimmunoassay was 7928Iu / mg (International Standard 6640) with cross-reaction (W / W) <0.5% of FSH (international standards <2%) and TSH <0.1% (international standards <1%). The purified human pituitary FSH and LH prepared in this study reached the level of similar products in the world and filled the domestic blank.