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用于毛细管电泳分析蛋白质的重氮树脂/羧基富勒烯自组装共价毛细管涂层及其与重氮树脂/氧化石墨烯涂层的比较

Self-assembled covalent capillary coating of diazoresin/carboxyl fullerene for analysis of proteins by capillary electrophoresis and a comparison with diazoresin/graphene oxide coating.

作者信息

Yu Bing, Shu Xi, Cong Hailin, Chen Xin, Liu Huwei, Yuan Hua, Chi Ming

机构信息

College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071, China; Laboratory for New Fiber Materials and Modern Textile, Growing Base for State Key Laboratory, Qingdao University, China.

College of Chemistry and Chemical Engineering, Qingdao University, Qingdao 266071, China.

出版信息

J Chromatogr A. 2016 Mar 11;1437:226-233. doi: 10.1016/j.chroma.2016.02.001. Epub 2016 Feb 3.

Abstract

Self-assembled and covalently linked capillary coatings of carboxyl fullerenes (C60-COOH) were prepared using photosensitive diazoresin (DR) as a coupling agent. Layer by layer (LBL) self-assembled DR/C60-COOH coatings based on ionic bonding was fabricated first on the inner surface of silica capillary, and subsequently converted into covalent bonding after treatment with UV light through a unique photochemistry reaction of DR. The covalently bonded coatings had the ability of suppressing protein adsorption on the inner surface of silica capillary, and thus the baseline separation of lysozyme (Lys), cytochrome c (Cyt-c), bovine serum albumin (BSA) and myoglobin (Mb) was achieved within 13min by using capillary electrophoresis (CE). The covalently linked DR/C60-COOH capillary coatings presented good chemical stability and repeatability. The reproducibility of the separation of proteins was less than 1%, 2.5%, and 3.5%, respectively, for run-to-run, day-to-day, capillary-to-capillary, respectively; and the RSD of migration time for the proteins are all less than 2.5% after a continuous 100 times running in a coating column. Compared with DR/graphene oxide (GO) coatings prepared by the same method, the DR/C60-COOH capillary coatings showed excellent protein separation performance due to a self-lubrication based anti-fouling mechanism. Because of the replacement of highly toxic and moisture sensitive silane coupling agent by DR in the covalent coating preparation, this method may provide an environmentally friendly and simple way to prepare the covalently coated capillaries for CE.

摘要

以光敏重氮树脂(DR)作为偶联剂,制备了羧基富勒烯(C60-COOH)自组装且共价连接的毛细管涂层。首先在石英毛细管内表面制备基于离子键的层层(LBL)自组装DR/C60-COOH涂层,随后通过DR独特的光化学反应经紫外光处理后转化为共价键。共价键合涂层具有抑制蛋白质吸附在石英毛细管内表面的能力,因此通过毛细管电泳(CE)在13分钟内实现了溶菌酶(Lys)、细胞色素c(Cyt-c)、牛血清白蛋白(BSA)和肌红蛋白(Mb)的基线分离。共价连接的DR/C60-COOH毛细管涂层具有良好的化学稳定性和重复性。蛋白质分离的重现性在连续运行、每日运行、毛细管间运行时分别小于1%、2.5%和3.5%;在涂层柱中连续运行100次后,蛋白质迁移时间的相对标准偏差均小于2.5%。与通过相同方法制备的DR/氧化石墨烯(GO)涂层相比,DR/C60-COOH毛细管涂层由于基于自润滑的防污机制而表现出优异的蛋白质分离性能。由于在共价涂层制备中用DR替代了剧毒且对湿度敏感的硅烷偶联剂,该方法可能为制备用于CE的共价涂层毛细管提供一种环境友好且简单的方法。

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