Murphy Sandra, Dowling Paul
Department of Biology, Maynooth University, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Methods Mol Biol. 2018;1664:109-114. doi: 10.1007/978-1-4939-7268-5_10.
The discovery of clinically relevant biomarkers using gel-based proteomics has proven extremely challenging, principally because of the large dynamic range of protein abundances in biofluids such as blood and the fact that only a small number of proteins constitute the vast majority of total blood protein mass. Various separation, depletion, enrichment, and quantitative developments coupled with improvements in gel-based protein quantification technologies, specifically difference gel electrophoresis (DIGE), have contributed to significant improvements in the detection and identification of lower abundance proteins. One of these enrichment technologies, Proteominer, will be the focus of this chapter. The Proteominer technology a utilizes hexapeptide bead library with huge diversity to bind and enrich low-abundance proteins but at the same time suppressing the concentration of high-abundance proteins in subsequent analysis.
利用基于凝胶的蛋白质组学发现具有临床相关性的生物标志物已被证明极具挑战性，主要原因在于生物流体（如血液）中蛋白质丰度的动态范围很大，而且只有少数蛋白质构成了总血液蛋白质质量的绝大部分。各种分离、去除、富集和定量技术的发展，再加上基于凝胶的蛋白质定量技术（特别是差异凝胶电泳，DIGE）的改进，显著提高了低丰度蛋白质的检测和鉴定水平。其中一种富集技术——蛋白质捕集磁珠（Proteominer），将成为本章的重点。蛋白质捕集磁珠技术利用具有巨大多样性的六肽珠文库来结合和富集低丰度蛋白质，同时在后续分析中抑制高丰度蛋白质的浓度。
