Azarkan Mohamed, Huet Joëlle, Baeyens-Volant Danielle, Looze Yvan, Vandenbussche Guy
Laboratoire de Chimie Générale (CP: 609), Faculté de Médecine, Université Libre de Bruxelles, Campus Erasme, 808, route de Lennik, B-1070 Bruxelles, Belgium.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Apr 15;849(1-2):81-90. doi: 10.1016/j.jchromb.2006.10.056. Epub 2006 Nov 17.
Separation or fractionation of a biological sample in order to reduce its complexity is often a prerequisite to qualitative or quantitative proteomic approaches. Affinity chromatography is an efficient protein separation method based on the interaction between target proteins and specific immobilized ligands. The large range of available ligands allows to separate a complex biological extract in different protein classes or to isolate the low abundance species such as post-translationally modified proteins. This method plays an essential role in the isolation of protein complexes and in the identification of protein-protein interaction networks. Affinity chromatography is also required for quantification of protein expression by using isotope-coded affinity tags.
为降低生物样品的复杂性而进行分离或分级分离,通常是定性或定量蛋白质组学方法的前提条件。亲和色谱法是一种基于目标蛋白与特定固定化配体之间相互作用的高效蛋白质分离方法。大量可用的配体能够将复杂的生物提取物分离成不同的蛋白质类别,或分离低丰度物种,如翻译后修饰的蛋白质。该方法在蛋白质复合物的分离以及蛋白质-蛋白质相互作用网络的鉴定中起着至关重要的作用。通过使用同位素编码亲和标签对蛋白质表达进行定量时,也需要亲和色谱法。