Electroosmotic pump-assisted capillary electrophoresis of proteins.
Electroosmotic pump-assisted capillary electrophoresis of proteins.
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DOI：
10.1016/j.chroma.2009.06.032
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发表时间：
2009-08
期刊：
影响因子：
--
通讯作者：
Liang Xu;Xiaoyan Dong;Y. Sun
中科院分区：
文献类型：
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作者：
Liang Xu;Xiaoyan Dong;Y. Sun
一种新的蛋白质分析方法，即电渗泵辅助毛细管电泳（EOPACE），开发和证明具有几个优势，比其他CE为基础的技术。EOPACE中使用的色谱柱由两个连接部分组成，即聚乙烯醇（PVA）涂层和未涂层毛细管。PVA涂层毛细管柱是EOPACE中蛋白质电泳的部分。电渗流（EPOF）几乎完全抑制在这个亲水性聚合物涂层部分，所以蛋白质电泳在PVA改性的毛细管是免费的不可逆的蛋白质吸附到毛细管内壁。未涂覆的毛细管部分用作电渗泵，因为在裸二氧化硅毛细管中在中性pH下发生朝向阴极的电迁移。通过对含有细胞色素c（Cyt-c）、肌红蛋白和胰蛋白酶抑制剂的蛋白质混合物的分离，证明了EOPACE方法相对于压力辅助CE、裸毛细管区带电泳（CZE）和PVA涂层毛细管区带电泳（CZE）的优越性。EOPACE的一个显著特征是在中性pH下同时分离阳离子、阴离子和不带电荷的蛋白质可以通过单次运行容易地完成，这是其他基于CE的方法不可能或难以实现的。对EOPACE在蛋白质分析中的高柱效和良好的重现性进行了验证和讨论。此外，证明了用EOACE系统分离Cyt-c的胰蛋白酶解物。
A new method for protein analysis, that is, electroosmotic pump-assisted capillary electrophoresis (EOPACE), is developed and demonstrated to possess several advantages over other CE-based techniques. The column employed in EOPACE consists of two linked sections, poly(vinyl alcohol) (PVA)-coated and uncoated capillaries. The PVA-coated capillary column is the section for protein electrophoresis in EOPACE. Electroosmotic flow (EOF) is almost completely suppressed in this hydrophilic polymer coated section, so protein electrophoresis in the PVA-modified capillary is free of irreversible protein adsorption to the capillary inner wall. The uncoated capillary section serves as an electroosmotic pump, since EOF towards cathode occurs at neutral pH in the naked silica capillary. By the separation of a protein mixture containing cytochrome c (Cyt-c), myoglobin and trypsin inhibitor, we have demonstrated the advantages of EOPACE method over other relevant ones such as pressure assisted CE, capillary zone electrophoresis (CZE) with naked capillary and CZE with PVA-coated capillary. A significant feature of EOPACE is that simultaneous separation of cationic, anionic and uncharged proteins at neutral pH can be readily accomplished by a single run, which is impossible or difficult to realize by the other CE-based methods. The high column efficiency and good reproducibility in protein analysis by EOPACE are verified and discussed. In addition, separation of tryptic digests of Cyt-c with the EOPACE system is demonstrated.