Organic solvent extraction of proteins and peptides from serum as an effective sample preparation for detection and identification of biomarkers by mass spectrometry.
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.
Chertov
Oleg
O
Protein Chemistry Laboratory, SAIC-Frederick, MD, USA. oleg@ncifcrf.gov
Biragyn
Arya
A
Kwak
Larry W
LW
Simpson
John T
JT
Boronina
Tatiana
T
Hoang
Van M
VM
Prieto
DaRue A
DA
Conrads
Thomas P
TP
Veenstra
Timothy D
TD
Fisher
Robert J
RJ
D000097:Acetonitriles / Q000737:chemistry*; D000818:Animals; D016633:Apolipoprotein A-II / Q000378:metabolism*; D015415:Biomarkers / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D016393:Lymphoma, B-Cell / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D010455:Peptides / Q000097:blood / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014269:Trifluoroacetic Acid / Q000737:chemistry*
D000097:Acetonitriles; D016633:Apolipoprotein A-II; D015415:Biomarkers; D010455:Peptides; D011506:Proteins; D014269:Trifluoroacetic Acid; C032159:acetonitrile