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Characterization of the human blood plasma proteome
Author(s) -
Shen Yufeng,
Kim Jeongkwon,
Strittmatter Eric F.,
Jacobs Jon M.,
Camp David G.,
Fang Ruihua,
Tolié Nikola,
Moore Ronald J.,
Smith Richard D.
Publication year - 2005
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200401246
Subject(s) - chromatography , proteome , blood proteins , chemistry , tandem mass spectrometry , albumin , mass spectrometry , proteomics , liquid chromatography–mass spectrometry , affinity chromatography , human serum albumin , biochemistry , gene , enzyme
We describe methods for broad characterization of the human plasma proteome. The combination of stepwise immunoglobulin G (IgG) and albumin protein depletion by affinity chromatography and ultrahigh‐efficiency capillary liquid chromatography separations coupled to ion trap‐tandem mass spectrometry enabled identification of 2392 proteins from a single plasma sample with an estimated confidence level of > 94%, and an additional 2198 proteins with an estimated confidence level of 80%. The relative abundances of the identified proteins span a range of over eight orders of magnitude in concentration (< 30 pg/mL to ∼30 mg/mL), facilitated by the attomole‐level sensitivity of the analysis methods. More than 80% of the observed proteins demonstrate interactions with IgG and/or albumin, and the human plasma protein loss in the affinity chromatography/strong cation exchange/reversed‐phase liquid chromatography‐tandem mass spectrometry methodology was investigated in detail. The results of this study provide a basis for a wide range of plasma proteomics studies, including broad quantitation of relative abundances in comparative studies of the identification of novel protein disease markers, as well as further studies of protein‐protein interactions.