Shen Yufeng, Smith Richard D
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
Electrophoresis. 2002 Sep;23(18):3106-24. doi: 10.1002/1522-2683(200209)23:18<3106::AID-ELPS3106>3.0.CO;2-Y.
Identifying and quantifying in a high throughput manner the proteins expressed by cells, tissues or an organism provides the basis for understanding the functions of its constituents at a "systems" level. As a result, proteome analysis has increasingly become the focus of significant interest and research over the past decade. This is especially true following the recent stunning achievements in genomics analyses. However, unlike the static genome, the complexities and dynamism of the proteome present significant analytical challenges and demand highly efficient separations and detection technologies. A number of recent technological advancements have been in direct response to these challenges. Currently, strategically mated combinations of sophisticated separations techniques and advanced mass spectrometric detection represent the best approach to addressing the intricacies of the proteome. Liquid-phase separations, often within capillaries, are increasingly recognized as the best separations technique for this approach. In combination on-line with mass spectrometry, liquid-phase separations provide the improved analytical sensitivity, sample throughput, and quantitation capabilities necessitated by the multifaceted problems within proteomics analyses. This review focuses primarily on current high-efficiency capillary separations techniques, including both capillary liquid chromatography and capillary electrophoresis, applied to the analysis of complex proteomic samples. We emphasize developments at our laboratory and illustrate technical advances that attempt to review the role of separations within the broader context of a state-of-the-art integrated proteomics effort.
以高通量方式识别和定量细胞、组织或生物体表达的蛋白质，为在“系统”层面理解其组成部分的功能提供了基础。因此，蛋白质组分析在过去十年中越来越成为人们极大关注和研究的焦点。在基因组学分析取得近期惊人成就之后，情况尤其如此。然而，与静态的基因组不同，蛋白质组的复杂性和动态性带来了重大的分析挑战，需要高效的分离和检测技术。最近的一些技术进步正是直接针对这些挑战而产生的。目前，复杂分离技术与先进质谱检测的策略性结合是应对蛋白质组复杂性的最佳方法。液相分离（通常在毛细管内）越来越被认为是这种方法中最佳的分离技术。与质谱在线联用，液相分离提供了蛋白质组学分析中多方面问题所需的更高分析灵敏度、样品通量和定量能力。本综述主要关注当前应用于复杂蛋白质组样品分析的高效毛细管分离技术，包括毛细管液相色谱和毛细管电泳。我们强调我们实验室的进展，并举例说明在最先进的综合蛋白质组学研究的更广泛背景下，试图审视分离作用的技术进步。
