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季铵化纤维素负载金纳米粒子作为毛细管涂层用于增强毛细管电泳中蛋白质分离

Quaternized cellulose-supported gold nanoparticles as capillary coatings to enhance protein separation by capillary electrophoresis.

机构信息

Department of Chemistry and Key Laboratory of Biomedical Polymers of Ministry of Education, Wuhan University, 430072, China.

Center for Disease Prevention and Control of Futian District, Shenzhen 518040, China.

出版信息

J Chromatogr A. 2014 May 23;1343:160-6. doi: 10.1016/j.chroma.2014.03.079. Epub 2014 Apr 4.

Abstract

Gold nanoparticles (Au NPs) were synthesized and stabilized by using water-soluble quaternized cellulose (QC) as support matrix through a straightforward and environmentally friendly aqueous-phase approach. The structure and morphology of QC-supported Au NPs (QC-Au NPs) were investigated systematically by UV-visible, FT-IR, x-ray diffraction and TEM measurement. The Au NPs with mean diameter of about 7nm were shown to efficiently redisperse in water due to the strong interaction between QC and Au NPs, and the solutions were quite stable after storage for nearly 4 months at room temperature. QC-Au NPs were subsequently used as novel physically adsorbed coatings for protein separation by CE. The separation performance was significantly improved in the capillary coated by QC-Au NPs compared with that of the uncoated capillary or QC coated capillary. A small quantity of Au NPs (Au content of 4.6%) was adequate for the obvious improvement of coating ability. The theoretical plate number of lysozyme in QC-Au1 NPs coated capillary was 2.9 times as much as that in QC coated capillary. We have demonstrated the separation of six model proteins with RSD of migration time less than 2.79% and RSD of peak area less than 4.81%. Furthermore, QC-Au NPs was applied to the analysis of closely related proteins and biological samples. With simplicity, high resolution and reproducibility, the proposed method shows potential for applications in proteomics and clinical diagnosis.

摘要

金纳米粒子(Au NPs)通过简单、环保的水相方法合成并稳定在水溶性季铵化纤维素(QC)上。通过紫外可见光谱、傅里叶变换红外光谱、X 射线衍射和透射电子显微镜测量系统地研究了 QC 负载 Au NPs(QC-Au NPs)的结构和形态。由于 QC 和 Au NPs 之间的强相互作用,平均直径约为 7nm 的 Au NPs 能够有效地在水中重新分散,并且在室温下储存近 4 个月后,溶液仍相当稳定。随后,QC-Au NPs 被用作 CE 中蛋白质分离的新型物理吸附涂层。与未涂层毛细管或 QC 涂层毛细管相比,涂有 QC-Au NPs 的毛细管的分离性能显著提高。少量的 Au NPs(Au 含量为 4.6%)就足以明显提高涂层能力。在 QC-Au1 NPs 涂层毛细管中溶菌酶的理论塔板数是 QC 涂层毛细管的 2.9 倍。我们已经证明了六种模型蛋白质的分离,迁移时间的 RSD 小于 2.79%,峰面积的 RSD 小于 4.81%。此外,QC-Au NPs 还应用于分析密切相关的蛋白质和生物样品。该方法具有简单、高分辨率和重现性,在蛋白质组学和临床诊断中具有应用潜力。

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