Separation of acidic and basic proteins by nanoparticle-filled capillary electrophoresis
Separation of acidic and basic proteins by nanoparticle-filled capillary electrophoresis
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DOI：
10.1021/ac061059c
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发表时间：
2006-12-01
影响因子：
7.4
通讯作者：
Tseng, Wei-Lung
中科院分区：
文献类型：
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作者：
Yu, Cheng-Ju;Su, Chih-Lin;Tseng, Wei-Lung
我们提出的第一个例子分析的酸性和碱性蛋白质的纳米粒子填充毛细管电泳。与双十二烷基二甲基溴化铵（DDAB）修饰的毛细管相比，DDAB修饰的金纳米粒子（AuNPs）作为假固定相能在毛细管壁上形成更稳定的涂层，从而提高了分离效率和重现性.除了它们用于蛋白质分离的优点之外，DDAB封端的AuNP可以产生高的反向电渗流，其在pH 3.5时比DDAB大75%。为了允许与蛋白质的强相互作用，AuNPs通过非共价键合用聚（环氧乙烷）修饰以形成金纳米颗粒/聚合物复合物（AuNPPs）。在酸性条件下（10 mM磷酸盐，pH3.5），用DDAB-封端的AuNPs动态包被并填充AuNPPs的毛细管对酸性和碱性蛋白质进行分离，峰效率范围为71000 ~ 1007000板/m，迁移时间的相对标准偏差小于0.6%。此外，所提出的方法已被应用于生物样品，包括唾液，红细胞和血浆的分析。该方法具有操作简单、分辨率高、重复性好等优点，在蛋白质组学和临床诊断中具有广阔的应用前景。
We present the first example of the analysis of acidic and basic proteins by nanoparticle- filled capillary electrophoresis. Compared to the didodecyldimethylammonium bromide (DDAB)-coated capillary, the DDAB-capped gold nanoparticles (AuNPs) as pseudostationary phase were found to form more stable coating on the capillary wall, thus leading to greater separation efficiency and high reproducibility. In addition to their advantages for protein separation, DDAB-capped AuNPs can generate high reversed electroosmotic flow, which is 75% greater than DDAB at pH 3.5. To allow strong interactions with proteins, the AuNPs were modified with poly( ethylene oxide) via noncovalent bonding to form gold nanoparticles/ polymer composites (AuNPPs). Using a capillary dynamically coated with DDAB-capped AuNPs and filled with AuNPPs under acidic conditions (10 mM phosphate, pH 3.5), we have demonstrated the separation of acidic and basic proteins with peak efficiencies ranging from 71 000 to 1 007 000 plates/m and relative standard deviations of migration time less than 0.6%. Additionally, the proposed method has been applied to the analyses of biological samples, including saliva, red blood cells, and plasma. With simplicity, high resolving power, and high reproducibility, the proposed method has shown great potential for proteomics applications and clinical diagnosis.