Zolotarjova Nina, Mrozinski Peter, Chen Haiying, Martosella James
Agilent Technologies Inc., 2850 Centerville Road, Wilmington, DE 19808, USA.
J Chromatogr A. 2008 May 2;1189(1-2):332-8. doi: 10.1016/j.chroma.2007.11.082. Epub 2007 Dec 4.
The tremendous complexity of the serum and plasma proteome presents extreme analytical challenges in comprehensive analysis due to the wide dynamic range of protein concentrations. Therefore, robust sample preparation methods remain one of the important steps in the proteome characterization workflow. We present the results on a new column for the specific depletion of 14 high-abundant proteins from human serum and plasma and the subsequent reversed-phase fractionation of the flow-through proteins. Analysis of tryptic peptides was accomplished with microfluidic HPLC-Chip/MS system. Results indicate that high-abundant protein depletion combined with RP fractionation of plasma showed an improved dynamic range for proteomic analysis and enabled the identification of low-abundant plasma proteins.
由于蛋白质浓度的动态范围很宽，血清和血浆蛋白质组的巨大复杂性给全面分析带来了极大的分析挑战。因此，稳健的样品制备方法仍然是蛋白质组表征工作流程中的重要步骤之一。我们展示了一种新型柱的相关结果，该柱可特异性去除人血清和血浆中的14种高丰度蛋白质，并对流出的蛋白质进行后续反相分级分离。用微流控HPLC-芯片/MS系统完成了胰蛋白酶肽段的分析。结果表明，高丰度蛋白质去除与血浆的反相分级分离相结合，提高了蛋白质组分析的动态范围，并能够鉴定低丰度血浆蛋白质。
