Journal
TALANTA
Volume 85, Issue 2, Pages 1001-1006Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2011.05.008
Keywords
Tryptophan-containing peptides; Selective enrichment; Hydrazide-functionalized magnetic microspheres; Mass spectrometry; Serum
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Funding
- National Nature Science Foundation of China [20735005, 30700990, 209750243]
- S973 program [2007CB914100, 2010CB912700]
- 863 program [2006AA02A308]
- Shanghai Leading Academic Discipline Grant [B109]
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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. (C) 2011 Published by Elsevier B.V.
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