Human plasma proteome analysis by multidimensional chromatography prefractionation and linear ion trap mass spectrometry identification
Human plasma proteome analysis by multidimensional chromatography prefractionation and linear ion trap mass spectrometry identification
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DOI：
10.1021/pr049761h
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发表时间：
2005-03-01
影响因子：
4.4
通讯作者：
Zeng, R
中科院分区：
文献类型：
--
作者：
Jin, WH;Dai, J;Zeng, R
近年来，人们对人类血浆蛋白质组的兴趣重新抬头，因为它在疾病诊断和治疗监测方面具有巨大的革命性前景。多维液相色谱作为最有效的分离技术之一，引起了人们的广泛关注，但大多数已发表的工作集中在胰蛋白酶肽的分级分离上。在这项研究中，从人血浆中的蛋白质预分级的在线连续强阳离子交换色谱和反相色谱。所得30个样品分别用胰蛋白酶消化，并通过毛细管反相液相色谱-线性离子阱质谱联用分析。在满足严格的标准后，我们的工作中总共成功鉴定了1292个不同的蛋白质，其中一些已知存在于血清中的蛋白质< 10 ng/mL。与已发表的文献相比，该分析提供了一个更全面的血浆蛋白质组列表。考虑到我们的策略允许高通量的血清蛋白质鉴定，在MS分析之前的蛋白质预分离是一种简单有效的方法，以促进人血浆蛋白质组研究。
A resurgence of interest in the human plasma proteome has occurred in recent years because it holds great promise of revolution in disease diagnosis and therapeutic monitoring. As one of the most powerful separation techniques, multidimensional liquid chromatography has attracted extensive attention, but most published works have focused on the fractionation of tryptic peptides. In this study, proteins from human plasma were prefractionated by online sequential strong cation exchange chromatography and reversed-phase chromatography. The resulting 30 samples were individually digested by trypsin, and analyzed by capillary reversed-phase liquid chromatography coupled with linear ion trap mass spectrometry. After meeting stringent criteria, a total of 1292 distinct proteins were successfully identified in our work, among which, some proteins known to be present in serum in < 10 ng/mL were detected. Compared with other works in published literatures, this analysis offered a more full-scale list of the plasma proteome. Considering our strategy allows high throughput of protein identification in serum, the prefractionation of proteins before MS analysis is a simple and effective method to facilitate human plasma proteome research.