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基于单分散亲水性无孔树脂的强阳离子交换填料的制备及其在蛋白质快速分离中的应用

[Preparation of strong cation exchange packings based on monodisperse hydrophilic non-porous resins and their application for fast separation of proteins].

作者信息

Zhu Jinxia, Bo Chunmiao, Gong Bolin

机构信息

Key Laboratory of Energy & Chemical Engineering, Ningxia University, Yinchuan 750021, China.

出版信息

Se Pu. 2006 Mar;24(2):129-34. doi: 10.1016/s1872-2059(06)60007-2.

Abstract

Monodisperse, 3.0 microm non-porous hydrophilic poly (glycidylmethacrylate-co-ethylenedimethacrylate) particles were prepared by an one-step swelling and polymerization method. The particles were modified to be a strong cation exchange (SCX) stationary phase for high performance liquid chromatography (HPLC) in the following steps. First, the particles were completely hydrolyzed. Second, the hydrolyzed particles were treated with epichlorhydrin followed by another hydrolysis of the newly introduced epoxide groups. Third, the particles were reacted with chlorosulfonic acid. The SCX stationary phase was evaluated in light of the ion exchange property, separability and hydrophilicity on the separation and retention of proteins in detail. Four proteins were quickly separated in 1.0 min with linear gradient elution using the synthesized SCX stationary phase. It was found that it followed ion exchange chromatographic (IEC) retention mechanism. The SCX resin was used for the fast purification of lysozyme from egg white and cytochrome-C from pig heart in 3.0 min with only one step. The results obtained were satisfactory.

摘要

通过一步溶胀聚合方法制备了单分散的、3.0微米的无孔亲水性聚(甲基丙烯酸缩水甘油酯 - 二甲基丙烯酸乙二酯)颗粒。通过以下步骤将这些颗粒改性为用于高效液相色谱(HPLC)的强阳离子交换(SCX)固定相。首先,将颗粒完全水解。其次,用环氧氯丙烷处理水解后的颗粒,然后对新引入的环氧基团进行另一次水解。第三,使颗粒与氯磺酸反应。从离子交换性能、可分离性和亲水性方面详细评估了SCX固定相在蛋白质分离和保留方面的性能。使用合成的SCX固定相通过线性梯度洗脱在1.0分钟内快速分离了四种蛋白质。发现其遵循离子交换色谱(IEC)保留机制。使用SCX树脂仅一步即可在3.0分钟内从蛋清中快速纯化溶菌酶,并从猪心中快速纯化细胞色素C。获得的结果令人满意。

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