【摘要】 本文报道一种HCG 及其亚单位的制备和纯化方法。正常早期妊娠妇女尿经苯甲酸吸附、50%乙醇抽提、0.3M 醋酸钠缓冲液(用50%乙醇配制,pH4.8)再抽提、低温酒精沉淀干燥后得粗制品。其生物学活性为2000-3000国际单位/毫克。粗制品经DEAE-纤维素柱层析和Sephadex G-100凝胶过滤两步纯化后,得生物学活性为12000国际单位/毫克以上的精制品。用尿素解离此精制品,再经Sephadex G-100凝胶过滤纯化,可得生物学活性很低的α及β亚单位。经重结合后可恢复70—82%的生物学活性。更多还原

【Abstract】 In the past decade,great interest has been aroused in studying human chorionic gona-dotropine(HCG),because its roles have been found in more and more scientific areas.Soit is necessary to obtain a highly purified preparation of HCG and its subunits for investiga-tion in this field.This paper presents a simple method used in our laboratory to preparesuch a kind of HCG and its subunits.To summarize,there are five steps in all for purifyingHCG and two steps for purifying its subunits.1.The method for the purification of HCG:(a)adsorption of HCG with benzoic acid from early pregnant woman’s urine;(b)extraction with 50% ethanol,pH 6.0;(c)reextraction with 0.3M sodium acetate-acetic acidbuffer,pH 4.8, in 50% ethanol;(d)ion exchange chromatography on DEAE-Cellulose and(e)gel filtration on Sephadex G-100.A product with a potency of 12,000-16,000 iu/mgexhibiting a broad band in polyacrylamide disc electrophoresis could be obtained.2.The method for the purification of subunits of HCG:(a)dissociation with 10M urea,pH 4.5,and separation on DEAE-Sephadex A-25;(b)purification on Sephasex G-100.The final products have a biological activity ofnot more than 10 iu/mg,exhibiting 2-3 and 4-5 bands in polyacrylamide disc gel electro-phoresis respectively in the case of Subunit α and Subunit β.更多还原