期刊
JOURNAL OF PROTEOMICS
卷 73, 期 3, 页码 689-695出版社
ELSEVIER
DOI: 10.1016/j.jprot.2009.10.013
关键词
Proteomics; Biomarkers; Label-free; NanoLC-MS/MS; LC-MSE; Serum; Plasma; IMAC; Immobilized metal affinity chromatography; SAX; Strong anion exchange; Proteins; Analysis
资金
- Stanley Medical Research Institute (USA)
- Psynova Neurotech LTD (UK)
In order to exploit human blood as a source of protein disease biomarkers, robust analytical methods are needed to overcome the inherent molecular complexity of this bio-fluid. We present the coupling of label-free SAX chromatography and IMAC to a data-independent nanoLC-MS/MS (nanoLC-MSE) platform for analysis of blood plasma and serum proteins. The methods were evaluated using protein standards added at different concentrations to two groups of samples. The results demonstrate that both techniques enable accurate protein quantitation using low sample volumes and a minimal number of fractions. Combining both methods, 883 unique proteins were identified, of which 423 proteins showed high reproducibility. The two approaches resulted in identification of unique molecular signatures with an overlap of approximately 30%, thus providing complimentary information on sub-proteomes. These methods are potentially useful for systems biology, biomarker discovery, and investigation of phosphoproteins in blood. (C) 2009 Elsevier B.V. All rights reserved.
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