Quantitative proteome analysis of human plasma following in vivo lipopolysaccharide administration using 16O/18O labeling and the accurate mass and time tag approach.
Identification of novel diagnostic or therapeutic biomarkers from human blood plasma would benefit significantly from quantitative measurements of the proteome constituents over a range of physiological conditions. Herein we describe an initial demonstration of proteome-wide quantitative analysis of human plasma. The approach utilizes postdigestion trypsin-catalyzed 16O/18O peptide labeling, two-dimensional LC-FTICR mass spectrometry, and the accurate mass and time (AMT) tag strategy to identify and quantify peptides/proteins from complex samples. A peptide accurate mass and LC elution time AMT tag data base was initially generated using MS/MS following extensive multidimensional LC separations to provide the basis for subsequent peptide identifications. The AMT tag data base contains >8,000 putative identified peptides, providing 938 confident plasma protein identifications. The quantitative approach was applied without depletion of high abundance proteins for comparative analyses of plasma samples from an individual prior to and 9 h after lipopolysaccharide (LPS) administration. Accurate quantification of changes in protein abundance was demonstrated by both 1:1 labeling of control plasma and the comparison between the plasma samples following LPS administration. A total of 429 distinct plasma proteins were quantified from the comparative analyses, and the protein abundances for 25 proteins, including several known inflammatory response mediators, were observed to change significantly following LPS administration.
Qian
Wei-Jun
WJ
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Monroe
Matthew E
ME
Liu
Tao
T
Jacobs
Jon M
JM
Anderson
Gordon A
GA
Shen
Yufeng
Y
Moore
Ronald J
RJ
Anderson
David J
DJ
Zhang
Rui
R
Calvano
Steve E
SE
Lowry
Stephen F
SF
Xiao
Wenzhong
W
Moldawer
Lyle L
LL
Davis
Ronald W
RW
Tompkins
Ronald G
RG
Camp
David G
DG
Smith
Richard D
RD
D001798:Blood Proteins / Q000032:analysis*; D002852:Chromatography, Ion Exchange; D002853:Chromatography, Liquid; D005583:Fourier Analysis; D006801:Humans; D008070:Lipopolysaccharides / Q000008:administration & dosage*; D010100:Oxygen / Q000378:metabolism*; D010103:Oxygen Isotopes; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
D001798:Blood Proteins; D008070:Lipopolysaccharides; D010103:Oxygen Isotopes; D020543:Proteome; D010100:Oxygen