Haselberg Rob, de Jong Gerhardus J, Somsen Govert W
Department of Biomedical Analysis, Utrecht University, 3508 TB Utrecht, The Netherlands.
J Chromatogr A. 2007 Aug 3;1159(1-2):81-109. doi: 10.1016/j.chroma.2007.05.048. Epub 2007 May 18.
Developments in the fields of protein chemistry, proteomics and biotechnology have increased the demand for suitable analytical techniques for the analysis of intact proteins. In 1989, capillary electrophoresis (CE) was combined with mass spectrometry (MS) for the first time and its potential usefulness for the analysis of intact (i.e. non-digested) proteins was shown. This article provides an overview of the applications of CE-MS within the field of intact protein analysis. The principles of the applied CE modes and ionization techniques used for CE-MS of intact proteins are shortly described. It is shown that separations are predominantly carried out by capillary zone electrophoresis and capillary isoelectric focusing, whereas electrospray ionization (ESI) and matrix-assisted laser desorption ionization (MALDI) are the most popular ionization techniques used for interfacing. The combination of CE with inductively coupled plasma (ICP) MS for the analysis of metalloproteins is also discussed. The various CE-MS combinations are systematically outlined and tables provide extensive overviews of the applications of each technique for intact protein analysis. Selected examples are given to illustrate the usefulness of the CE-MS techniques. Examples include protein isoform assignment, single cell analysis, metalloprotein characterization, proteomics and biomarker screening. Finally, chip-based electrophoresis combined with MS is shortly treated and some of its applications are described. It is concluded that CE-MS represents a powerful tool for the analysis of intact proteins yielding unique separations and information.
蛋白质化学、蛋白质组学和生物技术领域的发展增加了对适用于完整蛋白质分析的分析技术的需求。1989年，毛细管电泳（CE）首次与质谱（MS）联用，显示出其在完整（即未消化）蛋白质分析中的潜在用途。本文概述了CE-MS在完整蛋白质分析领域的应用。简要描述了用于完整蛋白质CE-MS的应用CE模式和电离技术的原理。结果表明，分离主要通过毛细管区带电泳和毛细管等电聚焦进行，而电喷雾电离（ESI）和基质辅助激光解吸电离（MALDI）是最常用的联用电离技术。还讨论了CE与电感耦合等离子体（ICP）MS联用分析金属蛋白的情况。系统地概述了各种CE-MS联用方式，并通过表格广泛介绍了每种技术在完整蛋白质分析中的应用。给出了一些选定的例子来说明CE-MS技术的实用性。例子包括蛋白质异构体鉴定、单细胞分析、金属蛋白表征、蛋白质组学和生物标志物筛选。最后，简要介绍了芯片电泳与MS联用的情况，并描述了其一些应用。结论是，CE-MS是分析完整蛋白质的有力工具，能实现独特的分离并提供信息。
