牛促卵泡素。天然激素及其α和β亚基的分离与特性鉴定。

Bovine follitropin. Isolation and characterization of the native hormone and its alpha and beta subunits.

Beckers J F, Closset J, Maghuin-Rogister G, Hennen G

Biochimie. 1977;59(10):825-31. doi: 10.1016/s0300-9084(77)80214-9.

Abstract

A reproducible procedure was developed for the purification of bovine follitropin. 2) The method involved ammonium sulfate precipitation, ion exchange and adsorption chromatography, concanacaline-A-Sepharose chromatography and gel filtration. 3) A specific radioligand receptor assay was used to monitor each chromatographical step. 4) The potency of highly purified bovine follitropin as measured by Steelman and Pohley bioassay was 62 times the NIH-FSH-B1 standard preparation. 5) Contaminations of bovine follitropin by other glycoprotein hormones such as thyrotropin and lutropin amounted to 3 and 0.45 per cent by weight respectively as measured by specific radioimmunoassays and radioligand receptor assays. 6) The subunits alpha and beta of bovine follitropin were obtained by incubation in acidic urea, the chains being then separated by anion exchange chromatography. The subunits were subjitted to complete characterization. The amino-terminal residue of the alpha subunit is phenylalanine while a half cystine residue was found at the aminoterminal end of the beta chain. 8) Cross-contamination of the alpha and beta subunit preparations was measured by specific radioimmunoassays and amounted to 0.02 and 0.1 per cent by weight respectively.

摘要

已开发出一种用于纯化牛促卵泡素的可重复程序。2) 该方法包括硫酸铵沉淀、离子交换和吸附色谱、伴刀豆球蛋白A-琼脂糖色谱和凝胶过滤。3) 使用特异性放射性配体受体测定法监测每个色谱步骤。4) 通过斯蒂尔曼和波赫利生物测定法测定，高度纯化的牛促卵泡素的效价是NIH-FSH-B1标准制剂的62倍。5) 通过特异性放射免疫测定法和放射性配体受体测定法测量，牛促卵泡素被其他糖蛋白激素如促甲状腺激素和促黄体激素的污染分别为重量的3%和0.45%。6) 牛促卵泡素的α和β亚基通过在酸性尿素中孵育获得，然后通过阴离子交换色谱分离这些链。对亚基进行了全面表征。α亚基的氨基末端残基是苯丙氨酸，而在β链的氨基末端发现了一个半胱氨酸残基。8) 通过特异性放射免疫测定法测量，α和β亚基制剂的交叉污染分别为重量的0.02%和0.1%。