Isolation of N-linked glycopeptides from plasma
Isolation of N-linked glycopeptides from plasma
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DOI：
10.1021/ac0623181
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发表时间：
2007-08-01
影响因子：
7.4
通讯作者：
Zhang, Hui
中科院分区：
文献类型：
--
作者：
Zhou, Yong;Aebersold, Ruedi;Zhang, Hui
血浆的蛋白质组学分析可以潜在地识别生物标志物，这些生物标志物可用于对个体的生理或病理状态进行分类以及监测治疗效果。然而，血浆蛋白质组的复杂性、每个蛋白质的动态蛋白质翻译后修饰产生的大量肽以及个体之间的序列变异对当前的蛋白质组技术提出了巨大的挑战。为了克服这些挑战，我们最近开发了一种利用 N-连接糖肽 (SPEG) 固相萃取对糖蛋白进行高通量分析的方法。在这里，我们描述了使用 SPEG 进行血浆分析的程序，其中 SPEG 的每个步骤都进行了优化。使用掺有放射性标记的人血浆糖蛋白的小鼠血浆来监测优化的性能。我们的数据表明，可以使用 SPEG 技术开发血浆蛋白质组分析的标准程序，这主要是由于血浆中蛋白质含量相对恒定。
Proteomic analysis of blood plasma can potentially identify biomarkers that are useful for classifying the physiological or pathological status of an individual and for monitoring the effects of therapy. However, the complexity of the plasma proteome, the large number of peptides generated per protein due to dynamic protein post-translational modifications of each protein, and sequence variations among individuals pose great challenges to current proteomic technologies. To overcome these challenges, we have recently developed a method for the high-throughput analysis of glycoproteins using solid-phase extraction of N-linked glycopeptides (SPEG). Here we describe a procedure for plasma analysis using SPEG in which each step of SPEG was optimized. The performance of optimization was monitored using mouse plasma spiked with radioactive-labeled human plasma glycoproteins. Our data show that a standard procedure for plasma proteome analysis can be developed using the SPEG technique, mainly due to the relatively constant protein content in plasma.