Improved proteomic discovery by sample pre-fractionation using dual-column ion-exchange high performance liquid chromatography
Improved proteomic discovery by sample pre-fractionation using dual-column ion-exchange high performance liquid chromatography
复制标题
DOI：
10.1016/j.jchromb.2006.10.075
复制
发表时间：
2007-02-15
影响因子：
3
通讯作者：
Emili, Andrew
中科院分区：
文献类型：
--
作者：
Havugimana, Pierre C.;Wong, Peter;Emili, Andrew
迫切需要临床相关的生物标志物来改善患者诊断、风险分层、预后和治疗。识别早期疾病的基于蛋白质的指标以进行更有效的干预有一个特别令人信服的动机。尽管最近取得了进展，但由于生物体液中纯粹的异质性和不平衡的蛋白质丰度，蛋白质组学的发现过程仍然是一项艰巨的挑战。相对于现代生物医学应用的需求，即使是最先进的质谱系统也表现出有限的整体动态范围和灵敏度。为此，我们报告了一种稳健、快速且可重复的高性能离子交换液相色谱预分级方法的开发，该方法允许使用基本串联质谱筛选程序提高复杂生物样本的蛋白质组检测覆盖率。与未分级的粗提物的直接分析相比，在全局蛋白质组分析之前进行的这种形式的样品简化（我们统称为“分级组学”）增加了可以在组织和细胞裂解物中自信地鉴定的蛋白质的数量和多样性。 (c) 2006 Elsevier B.V. 保留所有权利。
Clinically relevant biomarkers are urgently needed for improving patient diagnosis, risk stratification, prognosis and therapeutic treatments. There is a particularly compelling motivation for identifying protein-based indicators of early-stage disease for more effective interventions. Despite recent progress, the proteomic discovery process remains a daunting challenge due to the sheer heterogeneity and skewed protein abundances in biofluids. Even the most advanced mass spectrometry systems exhibit limiting overall dynamic ranges and sensitivities relative to the needs of modem biomedical applications. To this end, we report the development of a robust, rapid, and reproducible high performance ion-exchange liquid chromatography pre-fractionation method that allows for improved proteomic detection coverage of complex biological specimens using basic tandem mass spectrometry screening procedures. This form of sample simplification prior to global proteomic profiling, which we refer to collectively as 'fractionomics', increases the number and diversity of proteins that can be confidently identified in tissue and cell lysates as compared to the straight analysis of unfractionated crude extracts. (c) 2006 Elsevier B.V. All rights reserved.