Issaq Haleem J, Conrads Thomas P, Janini George M, Veenstra Timothy D
Analytical Chemistry Laboratory, SAIC Frederick NCI-Frederick, Frederick, MD 21702, USA.
Electrophoresis. 2002 Sep;23(17):3048-61. doi: 10.1002/1522-2683(200209)23:17<3048::AID-ELPS3048>3.0.CO;2-L.
In the last few years there has been an increased effort to develop technologies capable of identifying and quantifying large numbers of proteins expressed within a cell system (i.e., the proteome). The complexity of the mixtures being analyzed has made the development of effective fractionation and separation methods a critical component of this effort. This review highlights many of the protein and peptide fractionation and separation methods, such as electrophoresis and high-performance liquid chromatography (HPLC), which have experienced significant development over the past forty years. Modern instrumental strategies for the resolution of cell proteins, based on separations employing a single high-resolution or multidimensional approach, and the relative merits of each, will be discussed. The focus of this manuscript will be on the development of multidimensional separations such as two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), HPLC/HPLC, and HPLC-capillary electrophoresis and their application to the characterization of complex proteome mixtures.
在过去几年中，人们加大了开发能够识别和定量细胞系统中大量表达的蛋白质（即蛋白质组）的技术的力度。所分析混合物的复杂性使得开发有效的分级分离和分离方法成为这项工作的关键组成部分。本综述重点介绍了许多蛋白质和肽的分级分离和分离方法，如电泳和高效液相色谱（HPLC），这些方法在过去四十年中取得了显著进展。将讨论基于采用单一高分辨率或多维方法进行分离的现代细胞蛋白质分离仪器策略，以及每种方法的相对优点。本手稿的重点将是多维分离技术的发展，如二维聚丙烯酰胺凝胶电泳（2D-PAGE）、HPLC/HPLC和HPLC-毛细管电泳及其在复杂蛋白质组混合物表征中的应用。
