A study of glycoproteins in human serum and plasma reference standards (HUPO) using multilectin affinity chromatography coupled with RPLC-MS/MS.
The glycoproteome is a major subproteome present in human plasma. In this study, we isolated and characterized approximately 150 glycoproteins from the human plasma and serum samples provided by HUPO using a multilectin affinity column. The corresponding tryptic digest was separated by RP-HPLC coupled to an IT mass spectrometer (3-D LCQ). Also in this study, a new system, namely an Ettan MDLC system coupled to a linear ITLTQ, was compared with the previous LCQ platform and gave a greater number of protein identifications, as well as better quality. When we compared the composition of the glycoproteomes for the plasma and serum samples there was a close correlation between the samples, except for the absence of fibrinogen from the identified-protein list in the latter sample, which was presumably as a result of the clotting process. In addition, the analysis of the samples from three ethnic specimens, Caucasian American, Asian American, and African American, were very similar but showed a higher angiotensinogen plasma level and a lower histidine-rich glycoprotein level in Caucasian American samples, and a lower vitronectin level in African American blood samples.
Yang
Ziping
Z
Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA.
Hancock
William S
WS
Chew
Tori Richmond
TR
Bonilla
Leo
L
D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods / Q000592:standards*; D012015:Reference Standards
D001798:Blood Proteins; D006023:Glycoproteins; D037102:Lectins