快速弱阴离子交换无孔聚合物固定相的制备及用于蛋白质的高效液相色谱分离

Rapid Weak Anion-exchange Chromatography of Proteins on a Non-porous Polymeric Stationary Phase

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.