Department of Chemistry, Fudan University, Shanghai 200433, China.
Talanta. 2011 Aug 15;85(2):1001-6. doi: 10.1016/j.talanta.2011.05.008. Epub 2011 May 12.
The extreme complexity of protein samples is becoming a great challenge for proteomic analysis, especially for those having large dynamic range of protein abundance. To solve this problem, and to overcome the limitation of the current proteomic technologies, a new method using hydrazide-functionalized magnetic microspheres was established in this study. With this method, tryptophan (Trp)-containing peptides can be selectively and sensitively enriched from complex and low-volume samples. Furthermore, combined with 1D-LC-MS/MS analysis, the strategy was successfully applied to the proteomic study of mouse serum. The proportion of Trp-containing peptides was increased from 19.4% to 80.2% through enrichment, and the complexity of the sample was reduced more than two times. An additional 113 Trp-containing peptides and 48 novel proteins were detected compared to the conventional method. This enrichment method provides a means for identifying more proteins as potential biomarkers in serum and other complex samples.
蛋白质样品的极端复杂性正成为蛋白质组学分析的巨大挑战，特别是对于那些蛋白质丰度具有较大动态范围的样品。为了解决这个问题，并克服当前蛋白质组学技术的局限性，本研究建立了一种使用酰肼功能化磁性微球的新方法。利用该方法，可以从复杂和小体积的样品中选择性和灵敏地富集色氨酸（Trp）肽。此外，结合 1D-LC-MS/MS 分析，该策略成功应用于小鼠血清的蛋白质组学研究。通过富集，Trp 肽的比例从 19.4%增加到 80.2%，并且样品的复杂性降低了两倍以上。与常规方法相比，检测到 113 种新的 Trp 肽和 48 种新的蛋白质。这种富集方法为鉴定更多潜在的血清和其他复杂样品中的生物标志物蛋白提供了一种手段。
