色谱 ›› 2013, Vol. 31 ›› Issue (4): 310-316.DOI: 10.3724/SP.J.1123.2012.12005

• 特别策划:色谱固定相专栏 • 上一篇    下一篇

高亲水性强阳离子交换色谱填料的制备及其在蛋白质分析中的应用

刘吉众1,黄嫣嫣1,杨博2,常建华2,刘国诠1,赵睿1*   

  1. 1. 中国科学院化学研究所, 中国科学院活体分析化学重点实验室, 北京 100190;
    2. 西安交大保赛生物技术股份有限公司, 陕西 西安 710054
  • 收稿日期:2012-12-03 出版日期:2013-04-22 发布日期:2013-04-24
  • 通讯作者: Tel:(010)62557910,E-mail:zhaorui@iccas.ac.cn
  • 基金资助:

    国家自然科学基金项目(21075125,21105105).

Preparation of highly hydrophilic strong cation exchangers and their applications in protein analysis

LIU Jizhong1, HUANG Yanyan1, YANG Bo2, CHANG Jianhua2, LIU Guoquan1, ZHAO Rui1*   

  1. 1. CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China; 2. Xi’an Jiaoda Bio-Sep Technologies Co. Ltd, Xi’an 710054, China
  • Received:2012-12-03 Online:2013-04-22 Published:2013-04-24

摘要:

以具有双孔结构的聚甲基丙烯酸环氧丙酯(PGMA)微球为基质,以葡萄糖进行表面亲水改性,制备了强阳离子交换色谱填料,并将其用于复杂生命体系中生物大分子的快速而高效的分离、分析与纯化。葡萄糖亲水改性增进了填料的生物相容性,提高了蛋白质样品的回收率;双孔结构及较高的比表面积赋予填料良好的柱渗透性和样品负载量。以标准蛋白质为样品,考察了该填料对生物样品的分离性能。以100 mm×4.6 mm的色谱柱分离4种蛋白质,在6 min内实现了基线分离;以溶菌酶为样品,填料的吸附容量为39.5 g/L,在蛋白质快速分离纯化分析中显示了良好的应用前景。

关键词: 蛋白质分析, 固定相, 聚甲基丙烯酸环氧丙酯微球, 葡萄糖改性, 强阳离子交换

Abstract:

Based on the needs of new packing materials for rapid and efficient separation, purification and analysis of biomacromolecules, a novel sulfonic acid-type strong cation exchange resin (SP-G-PGMA SCX resin) was prepared. The porous poly(glycidyl methacrylate) microspheres (PGMA) were selected as the matrix and glucose was used as the hydrophilic modifier to block the hydrophobic domains of PGMA beads. Glucose modification on PGMA beads improved the biocompatibility and reduced the non-specific adsorption so as to increase the recoveries of protein. The PGMA beads possess the porous structure and the relatively high specific surface area, which make the PGMA-based resins good permeability and high loading capacity. The application of such SP-G-PGMA SCX resin for the chromatographic separation of biomacromolecules was explored. Four basic proteins were baseline separated within 6 min with the column size of 100 mm?4.6 mm. The adsorption capacity of lysozyme on SP-G-PGMA SCX resin was determined as 39.5 g/L. The results make the material promising for the separation and purification of biomacromolecules.

Key words: glucose modification, poly(glycidyl methacrylate) microspheres, protein analysis, stationary phase, strong cation-exchange

中图分类号: