从牛垂体中选择性提取生长激素及其进一步的纯化和结晶。

A selective extraction of growth hormone from bovine pituitary gland and its further purification and crystallization.

作者信息

Spitsberg V L

出版信息

Anal Biochem. 1987 Feb 1;160(2):489-95. doi: 10.1016/0003-2697(87)90081-9.

Abstract

A highly efficient method for the isolation of bovine growth hormone (GH) is described. The method is based on selective extraction of GH from 15,000 g subcellular sediment of anterior pituitary gland with 130-150 mM NH4HCO3, 1 mM EGTA, pH 7.2-7.4, at 2-6 degrees C for 60 min, purification of the extracted GH by ammonium sulfate fractionation, one-step ion-exchange column chromatography on DEAE-cellulose (or CM-cellulose), and gel filtration on Sephadex G-75. This 2-3 day procedure provides a highly pure hormone in high yield (up to 70-80 mg per 35-40 g of the whole pituitary gland), which can be crystallized by the batch method at a low ionic strength and isoelectric pH.

摘要

本文描述了一种高效分离牛生长激素（GH）的方法。该方法基于以下步骤：在2-6℃下，用130-150 mM碳酸氢铵、1 mM乙二醇双四乙酸（EGTA）、pH 7.2-7.4，从垂体前叶的15,000 g亚细胞沉淀物中选择性提取GH，提取60分钟；通过硫酸铵分级分离对提取的GH进行纯化；在DEAE-纤维素（或CM-纤维素）上进行一步离子交换柱色谱；以及在葡聚糖凝胶G-75上进行凝胶过滤。这个2-3天的过程能以高产率提供高纯度的激素（每35-40 g整个垂体可达70-80 mg），该激素可通过批量法在低离子强度和等电pH下结晶。