用于蛋白质纯化的聚苯乙烯基质的改性。聚乙烯醇吸附对用于亲和色谱的聚苯乙烯-二乙烯基苯珠粒特性的影响。

Modification of polystyrenic matrices for the purification of proteins. Effect of the adsorption of poly(vinyl alcohol) on the characteristics of poly(styrene-divinylbenzene) beads for use in affinity chromatography.

作者信息

Nash D C, McCreath G E, Chase H A

机构信息

Department of Chemical Engineering, University of Cambridge, UK.

出版信息

J Chromatogr A. 1997 Jan 10;758(1):53-64. doi: 10.1016/s0021-9673(96)00710-8.

DOI:10.1016/s0021-9673(96)00710-8

PMID:9181973

Abstract

A poly(styrene-divinylbenzene) (PSDVB) chromatography matrix, CG1000-sd (TosoHaas), has been modified using poly(vinyl alcohol) (PVA) to create a matrix suitable for the attachment of functional groups for the selective purification of proteins. The characteristics of the modified matrix have been studied using a BET nitrogen adsorption/desorption technique and it has been found that the adsorption of PVA results in the bead micropores being filled whilst the bead macropores are left essentially unaltered. There was no protein adsorption onto the modified matrices. A dye ligand (Procion Blue MX-R) has been covalently attached to PVA-PSDVB matrix and the lysozyme capacities of the PVA-PSDVB matrix have been determined. The matrix compares well with commercial Blue Sepharose Fast Flow, an affinity matrix on cross-linked agarose. The dye-PVA-PSDVB matrix is stable when subjected to sanitisation with sodium hydroxide.

摘要

一种聚（苯乙烯-二乙烯基苯）（PSDVB）色谱基质CG1000-sd（托索哈斯公司）已用聚乙烯醇（PVA）进行改性，以制备适合连接官能团用于蛋白质选择性纯化的基质。使用BET氮吸附/脱附技术研究了改性基质的特性，发现PVA的吸附导致珠粒微孔被填满，而珠粒大孔基本保持不变。改性基质上没有蛋白质吸附。一种染料配体（普施安蓝MX-R）已共价连接到PVA-PSDVB基质上，并测定了PVA-PSDVB基质的溶菌酶容量。该基质与交联琼脂糖上的亲和基质——商业蓝琼脂糖快速流动柱相比效果良好。用氢氧化钠进行消毒时，染料-PVA-PSDVB基质很稳定。