Reducing protein concentration range of biological samples using solid-phase ligand libraries
Reducing protein concentration range of biological samples using solid-phase ligand libraries
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DOI：
10.1016/j.jchromb.2005.12.048
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发表时间：
2006-03-20
影响因子：
3
通讯作者：
Boschetti, E
中科院分区：
文献类型：
--
作者：
Guerrier, L;Thulasiraman, V;Boschetti, E
从生物液体中发现具有诊断相关性的特异性多肽由于样品中所有多肽/蛋白质的总体大量和大浓度范围而变得复杂。耗尽或分馏方法已被用于选择性地去除丰富的蛋白质，但是，他们未能显着富集微量蛋白质。在这里，我们扩展了一种新的方法，该方法允许通过在单个简单步骤中同时稀释高丰度蛋白质和低丰度蛋白质来减少复杂混合物（如纯血清）中的蛋白质浓度范围。该方法利用大多样性的固相配体库。通过控制样品与配体的比例，可以调节蛋白质的相对浓度，使得通常不能通过经典分析方法检测到的大量肽或蛋白质变得容易检测。沿着对其他生物提取物的处理，具体描述了该方法用于降低未分级血清的动态范围的应用。分析表面增强激光解吸/电离质谱（SELDI-MS）技术和单和二维电泳（1-DE和2-DE）证明检测到的蛋白质的数量增加。将这种方法与另外的分级分离方法联系起来的实例证明，使用所谓的“自上而下”或“自下而上”的方法进行蛋白质组学分析，可检测物质的数量进一步增加。通过能够检测样品中更大比例的多肽/蛋白质，该方法可以显著有助于发现诊断相关的新生物标志物。(c)2006 Elsevier B. V.保留所有权利。
The discovery of specific polypeptides of diagnostic relevance from a biological liquid is complicated by the overall vast number and the large concentration range of all polypeptides/proteins in the sample. Depletion or fractionation methodologies have been used for selectively removing abundant proteins; however, they failed to significantly enrich trace proteins. Here we expand upon a new method that allows the reduction of the protein concentration range within a complex mixture, like neat serum, through the simultaneous dilution of high abundance proteins and the concentration of low abundance ones in a single, simple step. This methodology utilizes solid-phase ligand libraries of large diversity. With a controlled sample-to-ligand ratio it is possible to modulate the relative concentration of proteins such that a large number of peptides or proteins that are normally not detectable by classical analytical methods become, easily detectable. Application of this method for reducing the dynamic range of unfractionated serum is specifically described along with treatment of other biological extracts. Analytical surface enhanced laser desorption/ionization mass spectrometry (SELDI-MS) technology and mono- and two-dimensional electrophoresis (1-DE and 2-DE) demonstrate the increase in the number of proteins detected. Examples linking this approach with additional fractionation methods demonstrate a further increase in the number of detectable species using either the so-called "top down" or "bottom up" approaches for proteomics analysis. By enabling the detection of a greater proportion of polypeptides/proteins within a sample, this method may contribute significantly towards the discovery of new biomarkers of diagnostic relevance. (c) 2006 Elsevier B.V. All rights reserved.