Simpson David C, Smith Richard D
Pacific Northwest National Laboratory, Richland, WA, USA.
Electrophoresis. 2005 Apr;26(7-8):1291-305. doi: 10.1002/elps.200410132.
Mass spectrometry (MS)-based proteomics is currently dominated by the analysis of peptides originating either from digestion of proteins separated by two-dimensional gel electrophoresis (2-DE) or from global digestion; the simple peptide mixtures obtained from digestion of gel-separated proteins do not usually require further separation, while the complex peptide mixtures obtained by global digestion are most frequently separated by chromatographic techniques. Capillary electrophoresis (CE) provides alternatives to 2-DE for protein separation and alternatives to chromatography for peptide separation. This review attempts to elucidate how the most promising CE modes, capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF), might best be applied to MS-based proteomics. CE-MS interfacing, mass analyzer performance, column coating to minimize analyte adsorption, and sample stacking for CZE are considered prior to examining numerous applications. Finally, multidimensional systems that incorporate CE techniques are examined; CZE often finds use as a fast, final dimension before ionization for MS, while CIEF, being an equilibrium technique, is well-suited to being the first dimension in automated fractionation systems.
基于质谱(MS)的蛋白质组学目前主要通过分析源自二维凝胶电泳(2-DE)分离的蛋白质消化产物或整体消化的肽段来进行;从凝胶分离的蛋白质消化得到的简单肽混合物通常不需要进一步分离,而通过整体消化得到的复杂肽混合物最常通过色谱技术进行分离。毛细管电泳(CE)为蛋白质分离提供了替代2-DE的方法,为肽分离提供了替代色谱的方法。本综述试图阐明最有前景的CE模式,即毛细管区带电泳(CZE)和毛细管等电聚焦(CIEF),如何最好地应用于基于MS的蛋白质组学。在研究众多应用之前,先考虑了CE-MS接口、质量分析器性能、用于最小化分析物吸附的柱涂层以及CZE的样品堆积。最后,研究了结合CE技术的多维系统;CZE常用于在离子化用于MS之前作为快速的最后一维,而CIEF作为一种平衡技术,非常适合作为自动分级系统中的第一维。