Terent'ev A A, Moldogazieva N T, Tatarinov Iu S
Biull Eksp Biol Med. 1990 May;109(5):438-40.
Human alpha-fetoprotein (AFP) was isolated by affinity chromatography method on immobilized diethylstilbestrol from butanol extract of abortive material. Elution from the column was performed with 10% aqueous buffered butanol solution, pH 8.6. During one procedure human AFP-preparation containing about 10% of AFP and about 90% of albumin was obtained, with the yield about 60%. The preliminary incubation of extract of the abortive material with estrone raised AFP yield up to 85% with the increase of AFP content in the preparation up to 35%, and preincubation with estriol and estradiol caused the increase of the yield up to 88-92%, and AFP content in the preparation was 50% and 65%, respectively. The preincubation of human AFP with diethylstilbestrol lowers the yield of this protein, which testified to the possible binding of human AFP with free diethylstilbestrol; testosterone, hydrocortisone and desoxycorticosterone caused the increase of the yield of AFP. So the competitive variant of the affinity chromatography on immobilized diethylstilbestrol makes it possible to raise human AFP preparation purity and yield by decreasing the competition between AFP, and not binding free steroid hormones, ad albumin for immobilized diethylstilbestrol.
采用固定化己烯雌酚亲和层析法从流产物质的丁醇提取物中分离人甲胎蛋白。用pH 8.6的10%缓冲丁醇水溶液进行柱洗脱。在一个操作过程中，获得了含约10%甲胎蛋白和约90%白蛋白的人甲胎蛋白制剂，产率约为60%。流产物质提取物与雌酮预孵育可使甲胎蛋白产率提高至85%，制剂中甲胎蛋白含量提高至35%；与雌三醇和雌二醇预孵育可使产率分别提高至88% - 92%，制剂中甲胎蛋白含量分别为50%和65%。人甲胎蛋白与己烯雌酚预孵育会降低该蛋白的产率，这证明人甲胎蛋白可能与游离己烯雌酚结合；睾酮、氢化可的松和脱氧皮质酮可使甲胎蛋白产率提高。因此，固定化己烯雌酚亲和层析的竞争变体通过减少甲胎蛋白、不结合游离甾体激素和白蛋白与固定化己烯雌酚之间的竞争，提高人甲胎蛋白制剂的纯度和产率成为可能。
