Simultaneous Enrichment of Plasma Soluble and Extracellular Vesicular Glycoproteins Using Prolonged Ultracentrifugation-Electrostatic Repulsion-hydrophilic Interaction Chromatography (PUC-ERLIC) Approach
Esther Sok Hwee Cheow, Kae Hwan Sim, Dominique de Kleijn, Chuen Neng Lee, Vitaly Sorokin and Siu Kwan Sze
Molecular & Cellular Proteomics (2015) 14: 1657-1671.
The authors investigated circulating glycoproteins and extracellular vesicles secreted from damaged or infected host cells and tissues, potential sources of biomarkers of human pathologies. The study showed that glycoprotein enrichment by PUC-ERLIC facilitates the simultaneous recovery of both secretory and extracellular vesicle-derived glycoproteins from non-depleted human plasma.
In a single LC-MS/MS run, the authors identified 127 glycoproteins in human plasma samples, including 599 unique glycopeptides and 361 unique glycosylation sites. The PUC-ERLIC method enabled the detection of 48 low-abundant glycoproteins (pg/ml-ng/ml) in human plasma without the need for prior depletion of high-abundance molecules. Using plasma obtained from heart disease patients, they identified, LYVE1, a cardiac-specific remodeling glycoprotein that has been reported to be involved in myocardium healing.