Proteomic analysis of human blood serum using peptide library beads
Proteomic analysis of human blood serum using peptide library beads
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DOI：
10.1021/pr070339l
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发表时间：
2007-10-01
影响因子：
4.4
通讯作者：
Rappsilber, Juri
中科院分区：
文献类型：
--
作者：
Sennels, Lau;Salek, Mogjilborahman;Rappsilber, Juri
人血清被认为含有诊断人类疾病的关键信息。然而，目前没有单一的技术，也可能永远无法应付血清蛋白质组的复杂性和动态范围。我们在此报告了一项大规模的蛋白质组学研究的人血清使用肽库珠和质谱。血清蛋白被吸附在涂有数百万个六聚体肽诱饵的组合文库的聚合物珠上。通过液相色谱法和高分辨率质谱法对该组合文库中的洗脱液（由3种不同强度的洗脱液获得，能够释放99%的保留物）进行分析，分别鉴定出1559种蛋白质或3869种蛋白质，这取决于95%置信度的估计。在任何一种情况下，分析表明配体珠能够在一次操作中捕获大量蛋白质。由于原始混合物中蛋白质浓度的“正常化”，与起始材料相比，配体头结合部分似乎具有较低的动态范围。我们发现，广泛的信息，蛋白质组成的复杂样品，如血清可以获得使用配体珠和这些珠丰富的蛋白质组学工具箱。
Human serum is thought to contain key information for diagnostics of human disease. However, no single technology is currently nor might ever be available to cope with the complexity and dynamic range of the serum proteome. We here report a large-scale proteomic study of human blood serum using peptide library beads and mass spectrometry. Serum proteins are adsorbed onto polymeric beads coated with a combinatorial library composed of millions of hexameric peptide baits. Analysis of the eluates from this combinatorial library (as obtained with 3 eluants of different strength, able to release 99% of the retentate) via liquid chromatography coupled to high-resolution mass spectrometry resulted in the identification of 1559 proteins or 3869 proteins, respectively, depending on how 95% confidence was estimated. In either case, the analysis showed that ligand beads are able to capture a large number of proteins in a single operation. The ligand bead bound fraction appeared to have a lower dynamic range when compared to the starting material, due to a "normalization" of the protein concentrations in the original mixture. We find that extensive information on the protein composition of complex samples such as serum can be obtained using ligand beads and that these beads enrich the proteomic tool box.