Characterization of the human blood plasma proteome
Characterization of the human blood plasma proteome
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DOI：
10.1002/pmic.200401246
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发表时间：
2005-10-01
期刊：
影响因子：
3.4
通讯作者：
Smith, RD
中科院分区：
文献类型：
--
作者：
Shen, YF;Kim, J;Smith, RD
我们描述了人类血浆蛋白质组的广泛特征的方法。通过亲和层析逐步分离免疫球蛋白和白蛋白，结合超高效毛细管液相色谱分离和离子捕获-串联质谱仪，可以从单个血浆样本中鉴定2392个蛋白质，估计置信度为94%，以及另外2198个蛋白质，估计置信度为80%。由于分析方法的全摩尔级灵敏度，鉴定出的蛋白质的相对丰度在浓度范围内超过八个数量级(<；30微克/毫升至类似于30毫克/毫升)。超过80%的观察到的蛋白质与免疫球蛋白和/或白蛋白相互作用，并详细研究了亲和层析/强阳离子交换/反相液相色谱质谱联用法中人血浆蛋白的损失。这项研究的结果为广泛的血浆蛋白质组学研究提供了基础，包括在识别新的蛋白质疾病标志物的比较研究中广泛定量的相对丰度，以及进一步的蛋白质-蛋白质相互作用的研究。
We describe methods for broad characterization of the human plasma proteome. The combination of stepwise immunoglobulin G (IgG) and albumin protein depletion by affinity chromatography and ultrahigh-efficiency capillary liquid chromatography separations coupled to ion trap-tandem mass spectrometry enabled identification of 2392 proteins from a single plasma sample with an estimated confidence level of > 94%, and an additional 2198 proteins with an estimated confidence level of 80%. The relative abundances of the identified proteins span a range of over eight orders of magnitude in concentration (< 30 mu g/mL to similar to 30 mg/mL), facilitated by the attomole-level sensitivity of the analysis methods. More than 80% of the observed proteins demonstrate interactions with IgG and/or albumin, and the human plasma protein loss in the affinity chromatography/strong cation exchange/reversed-phase liquid chromatographytandem mass spectrometry methodology was investigated in detail. The results of this study provide a basis for a wide range of plasma proteornics studies, including broad quantitation of relative abundances in comparative studies of the identification of novel protein disease markers, as well as further studies of protein-protein interactions.