一种纯化人T细胞生长因子的有效方法。

An efficient method for purification of human T-cell growth factor.

作者信息

Acuto O, Cianfriglia M, Colombatti M, Chapuis B, Nabholz M

出版信息

J Immunol Methods. 1982 Aug 27;53(1):15-26. doi: 10.1016/0022-1759(82)90236-8.

Abstract

A 1000-fold purification of human T-cell growth factor (TCGF) was achieved starting from supernatants of human spleen cells stimulated with phytohaemagglutinin (PHA) in culture medium containing 0.5% serum. The purification scheme involved precipitation with ammonium sulphate, gel filtration and blue-Sepharose chromatography. The use of polyethylene glycol 6000 (PEG 6000) was critical during the chromatographic steps in order to obtain high final recoveries or activity (40-50%). Purified preparations of TCGF labelled with 125I by the chloramine T method revealed that the activity co-migrated with 2 molecular species of 14,000-17,000 daltons in SDS-PAGE under non-reducing conditions.

摘要

从在含0.5%血清的培养基中用植物血凝素（PHA）刺激的人脾细胞上清液开始，实现了人T细胞生长因子（TCGF）1000倍的纯化。纯化方案包括硫酸铵沉淀、凝胶过滤和蓝色琼脂糖凝胶色谱。为了获得高的最终回收率或活性（40%-50%），在色谱步骤中使用聚乙二醇6000（PEG 6000）至关重要。通过氯胺T法用125I标记的TCGF纯化制剂显示，在非还原条件下的SDS-PAGE中，活性与两种分子量为14,000-17,000道尔顿的分子物种共同迁移。