摘要
将粒径为3.0μm无孔单分散亲水性交联聚甲基丙烯酸环氧丙酯树脂(PGMA/EDMA)的表面经不同的化学方法改性,制备了两种不同配基的弱阴离子交换(WAX-Ⅰ和WAX-Ⅱ)色谱填料。在同等色谱条件下,比较了两种填料对蛋白质的分离性能,发现WAX-Ⅰ型填料分离性能优于WAX-Ⅱ。详细考察了WAX-Ⅰ型弱阴离子交换色谱填料流动相pH值、流速及有机溶剂等对蛋白质保留的影响。实验结果表明,当流动相流速为3mL/min时,4种标准蛋白可在2min内基线快速分离,蛋白质的保留符合阴离子交换色谱规律。该填料可以用于生物工程产品的快速分离和纯化。
Two kinds of weak anion-exchange ( WAX-Ⅰ and WAX-Ⅱ ) stationary phases have been synthesized by chemical modification of the 3.0 μm non-porous monodisperse poly ( glycidylmethacrylate-co-ethylenedimethacrylate) (PGMA/EDMA) resin as medium. Under the same chromatographic conditions, compared the separation properties of the two kinds of WAX stationary phases for proteins, the results show that WAX-Ⅰ is much better than WAX-Ⅱ. The WAX-Ⅰ stationary phase was evaluated in detail to determine its ion-exchange properties, the effects of pH, flow rate and organic solvent on the separation and retention of the proteins. Four proteins could be separated in 2 min at flow rate of 3 mL/min with linear gradient 5 min elution by using the synthesized WAX-Ⅰ resin. It was found to have an anion-exchange chromatographic retention mechanism. The WAX-Ⅰ resin can be used for the fast separation and purification of bioengineering products.
出处
《分析测试学报》
CAS
CSCD
北大核心
2007年第1期8-11,15,共5页
Journal of Instrumental Analysis
基金
科技部基础研究前期专项基金(2004CC05000)
国家自然科学基金资助项目(20465001)
教育部新世纪优秀人才支持计划资助(NCET-04-0986)
关键词
无孔单分散交联聚甲基丙烯酸环氧丙酯树脂
弱阴离子交换色谱
固定相
蛋白质分离
Non-porous monodiperse poly ( glycidylmethacrylate-co-ethylenedimethacrylate ) beads
Weak anion-exchange chromatography
Stationary phase
Protein separation