Heparin chromatography to deplete high-abundance proteins for serum proteomics.
BACKGROUND
Serum is a very informative sample for disease diagnosis. However, a few of the high-abundance proteins existing in serum make the identification of disease-specific serum biomarkers extremely challenging using currently available technologies. A highly promising first step for most analytical approaches of serum is to deplete as many of the high-abundance proteins as possible.


METHODS
We introduced the traditional method of heparin chromatography coupled with protein G sepharose to deplete the high-abundance proteins for serum proteomics.


RESULTS
Compared with the multiple affinity removal system (MARS) column (a commercial version to deplete 6 major proteins in serum), heparin chromatography can deplete more high-abundance proteins in a single step, especially many high molecular-weight proteins. Using this simple and inexpensive method to pretreat serum for 2-DE analysis, more protein spots can be visualized. IgGs depletion by protein G sepharose can further enhance the resolution of the resulting serum proteome.


CONCLUSIONS
Heparin chromatography coupled with protein G appears to be an efficient and economical strategy to pretreat serum for serum proteomics.
Lei
Ting
T
Institute of Cancer Research, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, China.
He
Qing-Yu
QY
Wang
Yi-Li
YL
Si
Lu-Sheng
LS
Chiu
Jen-Fu
JF
D001798:Blood Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012680:Sensitivity and Specificity
D001798:Blood Proteins; D007074:Immunoglobulin G; D020543:Proteome; D006493:Heparin