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一种用于毛细管电泳分析蛋白质的新型重氮树脂/聚(N-乙烯基氨基丁酸)共价毛细管涂层。

A novel diazoresin/poly(N-vinyl aminobutyric acid) covalent capillary coating for the analysis of proteins by capillary electrophoresis.

作者信息

Yu Bing, Jiao Mingming, Cong Hailin, Shu Xi, Yang Shujing

机构信息

College of Chemical Engineering, Qingdao University, Qingdao, China; Laboratory for New Fiber Materials and Modern Textile - The Growing Base for State Key Laboratory, Qingdao University, China.

出版信息

J Sep Sci. 2014 Mar;37(6):725-30. doi: 10.1002/jssc.201301117. Epub 2014 Feb 19.

Abstract

A novel method for the preparation of covalently linked capillary coatings of poly(N-vinyl aminobutyric acid) (PVAA) obtained from hydrolyzed polyvinylpyrrolidone was demonstrated using photosensitive diazoresin (DR) as a coupling agent. A layer-by-layer self-assembled film of DR and PVAA based on ionic bonding was first fabricated on the inner wall of capillary, then ionic bonding was converted into covalent bonding after treatment with UV light through a unique photochemical reaction of DR. The covalently bonded coatings suppressed protein adsorption on the inner surface of the capillary, and thus a baseline separation of lysozyme, cytochrome c, BSA, amyloglucosidase, and myoglobin was achieved using CE. Compared with bare capillary or noncovalently bonded DR/PVAA coatings, the covalently linked DR/PVAA capillary coatings not only improved the CE separation performance for proteins, but also exhibited good stability and repeatability. Due to the replacement of the highly toxic and moisture-sensitive silane coupling agent by DR in the covalent coating preparation, this method may provide a green and easy way to make covalently coated capillaries for CE.

摘要

展示了一种制备共价连接的聚(N - 乙烯基氨基丁酸)(PVAA)毛细管涂层的新方法,该PVAA由水解的聚乙烯吡咯烷酮制得,使用光敏重氮树脂(DR)作为偶联剂。首先基于离子键在毛细管内壁制备DR和PVAA的逐层自组装膜,然后通过DR独特的光化学反应经紫外光处理后将离子键转化为共价键。共价键合涂层抑制了蛋白质在毛细管内表面的吸附,因此使用毛细管电泳实现了溶菌酶、细胞色素c、牛血清白蛋白、淀粉葡萄糖苷酶和肌红蛋白的基线分离。与裸毛细管或非共价键合的DR/PVAA涂层相比,共价连接的DR/PVAA毛细管涂层不仅提高了蛋白质的毛细管电泳分离性能,还表现出良好的稳定性和重复性。由于在共价涂层制备中用DR替代了剧毒且对水分敏感的硅烷偶联剂,该方法可能为制备用于毛细管电泳的共价涂层毛细管提供一种绿色且简便的方法。

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