Quantitative Profiling of Proteins in Complex Mixtures Using Liquid Chromatography and Mass Spectrometry
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- 6 June 2002
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of Proteome Research
- Vol. 1 (4) , 317-323
- https://doi.org/10.1021/pr025517j
Abstract
The objective of this study was to determine if liquid chromatography mass spectrometry (LC/MS) data of tryptic digests of proteins can be used for quantitation. In theory, the peak area of peptides should correlate to their concentration; hence, the peak areas of peptides from one protein should correlate to the concentration of that particular protein. To evaluate this hypothesis, different amounts of tryptic digests of myoglobin were analyzed by LC/MS in a wide range between 10 fmol and 100 pmol. The results show that the peak areas from liquid chromatography mass spectrometry correlate linearly to the concentration of the protein (r2 = 0.991). The method was further evaluated by adding two different concentrations of horse myoglobin to human serum. The results confirm that the quantitation method can also be used for quantitative profiling of proteins in complex mixtures such as human sera. Expected and calculated protein ratios differ by no more than 16%. We describe a new method combining protein identification with accurate profiling of individual proteins. This approach should provide a widely applicable means to compare global protein expression in biological samples. Keywords: mass spectrometry • protein quantification • protein identificationKeywords
This publication has 13 references indexed in Scilit:
- Differential stable isotope labeling of peptides for quantitation and de novo sequence derivationRapid Communications in Mass Spectrometry, 2001
- Quantitative Analysis of Bacterial and Mammalian Proteomes Using a Combination of Cysteine Affinity Tags and 15N-Metabolic LabelingAnalytical Chemistry, 2001
- Large-scale analysis of the yeast proteome by multidimensional protein identification technologyNature Biotechnology, 2001
- Towards defining the urinary proteome using liquid chromatography-tandem mass spectrometry I.Profiling an unfractionated tryptic digestProteomics, 2001
- Mechanistic investigation of ionization suppression in electrospray ionizationPublished by American Chemical Society (ACS) ,2000
- Quantitation of peptides and proteins by matrix-assisted laser desorption/ionization mass spectrometry using18O-labeled internal standardsRapid Communications in Mass Spectrometry, 2000
- Mass spectrometic detection for capillary isoelectric focusing separations of complex protein mixturesElectrophoresis, 2000
- Proteome studies of Saccharomyces cerevisiae: Identification and characterization of abundant proteinsElectrophoresis, 1997
- Linking genome and proteome by mass spectrometry: Large-scale identification of yeast proteins from two dimensional gelsProceedings of the National Academy of Sciences, 1996
- Two‐dimensional gel protein database of Saccharomyces cerevisiaeElectrophoresis, 1996