Sennels Lau, Salek Mogjiborahman, Lomas Lee, Boschetti Egisto, Righetti Pier Giorgio, Rappsilber Juri
The FIRC Institute for Molecular Oncology Foundation, Via Adamello 16, I-20139 Milan, Italy.
J Proteome Res. 2007 Oct;6(10):4055-62. doi: 10.1021/pr070339l. Epub 2007 Sep 18.
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.
人们认为人血清包含人类疾病诊断的关键信息。然而，目前没有一种技术能够应对血清蛋白质组的复杂性和动态范围，而且可能永远也不会有。我们在此报告一项使用肽库磁珠和质谱对人血清进行的大规模蛋白质组学研究。血清蛋白被吸附到涂有由数百万个六聚体肽诱饵组成的组合文库的聚合物磁珠上。通过液相色谱与高分辨率质谱联用对该组合文库的洗脱液（用三种不同强度的洗脱液获得，能够释放99%的保留物）进行分析，根据95%置信度的估计方式不同，分别鉴定出了1559种蛋白质或3869种蛋白质。无论哪种情况，分析都表明配体磁珠能够在一次操作中捕获大量蛋白质。与起始材料相比，配体磁珠结合部分的动态范围似乎较低，这是由于原始混合物中蛋白质浓度的“归一化”。我们发现，使用配体磁珠可以获得关于血清等复杂样品蛋白质组成的广泛信息，并且这些磁珠丰富了蛋白质组学工具箱。
