Organic solvent extraction of proteins and peptides from serum as an effective sample preparation for detection and identification of biomarkers by mass spectrometry

被引:129
作者
Chertov, O
Biragyn, A
Kwak, LW
Simpson, JT
Boronina, T
Hoang, VM
Prieto, DA
Conrads, TP
Veenstra, TD
Fisher, RJ
机构
[1] SAIC Frederick, Prot Chem Lab, Frederick, MD USA
[2] SAIC Frederick, Mass Spect Ctr, Frederick, MD USA
关键词
apolipoprotein A-II; biomarkers; serum; surface-enhanced laser desorption; ionization; tumor;
D O I
10.1002/pmic.200300677
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A method to extract peptides and low molecular weight proteins from serum under denaturing conditions using acetonitrile containing 0.1 % trifluoroacetic acid has been developed. The extraction procedure precipitates large, abundant proteins to simplify subsequent mass spectral analysis. This sample preparation method provides an efficient way to extract serum peptides, enabling them to be compared and identified using different mass spectrometry approaches. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry analysis of mouse blood serum samples prepared by this method allowed detection of two markers which were significantly reduced in mice with B cell lymphoma tumor. One of these markers has been identified as apolipoprotein A-II.
引用
收藏
页码:1195 / 1203
页数:9
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