Please use this identifier to cite or link to this item:
https://hdl.handle.net/10356/81887
Title: | Improving Blood Plasma Glycoproteome Coverage by Coupling Ultracentrifugation Fractionation to Electrostatic Repulsion–Hydrophilic Interaction Chromatography Enrichment | Authors: | Adav, Sunil S. Hwa, Ho Hee de Kleijn, Dominique Sze, Siu Kwan |
Keywords: | ERLIC; glycoproteins; plasma proteome; protein glycosylation; biomarkers | Issue Date: | 2015 | Source: | Adav, S. S., Hwa, H. H., de Kleijn, D.,& Sze, S. K. (2015). Improving Blood Plasma Glycoproteome Coverage by Coupling Ultracentrifugation Fractionation to Electrostatic Repulsion–Hydrophilic Interaction Chromatography Enrichment. Journal of Proteome Research, 14(7), 2828-2838. Adav, S. S., Hwa, H. H., de Kleijn, D., & Sze, S. K. (2015). Improving Blood Plasma Glycoproteome Coverage by Coupling Ultracentrifugation Fractionation to Electrostatic Repulsion–Hydrophilic Interaction Chromatography Enrichment. Journal of Proteome Research, 14(7), 2828-2838. |
Series/Report no.: | Journal of Proteome Research | Abstract: | Blood plasma is considered to be an excellent source of disease biomarkers because it contains proteins, lipids, metabolites, cell, and cell-derived extracellular vesicles from different cellular origins including diseased tissues. Most secretory and membranous proteins that can be found in plasma are glycoproteins; therefore, the plasma glycoproteome is one of the major subproteomes that is highly enriched with disease biomarkers. As a result, the glycoproteome has attracted much attention in clinical proteomic research. The modification of proteins with glycans regulates a wide range of functions in biology, but profiling plasma glycoproteins on a global scale has been hampered by the presence of low stoichiometry of glycoproteins in a complex high abundance plasma proteome background and lack of effective analytical technique. This study aims to improve plasma glycoproteome coverage using pig plasma as a model sample with a two-step strategy. The first step involves fractionation of the plasma proteins using ultracentrifugation into supernatant and pellet that is believed to contain low abundant glycoproteins. In the second step, further enrichment of glycopeptides was achieved in both fractions by adopting electrostatic repulsion hydrophilic interaction chromatography (ERLIC) coupled to tandem mass spectrometry (LC-MS/MS) analysis. The coverage of enriched glycoproteins in supernatant, pellet, and whole plasma sample as control was compared. Using this simple sample fractionation approach by ultracentrifugation and further ERLIC enrichment technique, sample complexity was reduced and glycoproteome coverage was significantly enhanced in supernatant and pellet fractions (by >50%) compared with whole plasma sample. This study showed that when ultracentrifugation is coupled to ERLIC glycopeptides enrichment and glycoproteome identification are significantly improved. This study demonstrates the combination of ultracentrifugation and ERLIC as a useful method for discovering plasma glycoprotein disease biomarkers. | URI: | https://hdl.handle.net/10356/81887 http://hdl.handle.net/10220/39735 |
ISSN: | 1535-3893 | DOI: | 10.1021/acs.jproteome.5b00102 | Schools: | School of Biological Sciences | Rights: | © 2015 American Chemical Society. This is the author created version of a work that has been peer reviewed and accepted for publication by Journal of Proteome Research, American Chemical Society. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1021/acs.jproteome.5b00102]. | Fulltext Permission: | open | Fulltext Availability: | With Fulltext |
Appears in Collections: | SBS Journal Articles |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Improving blood plasma glycoproteome coverage by coupling ultracentrifugation fractionation to electrostatic repulsion-hydrophilic interaction chromat.pdf | 855.79 kB | Adobe PDF | View/Open |
SCOPUSTM
Citations
20
14
Updated on Mar 21, 2024
Web of ScienceTM
Citations
20
14
Updated on Oct 31, 2023
Page view(s) 50
603
Updated on Mar 27, 2024
Download(s) 20
233
Updated on Mar 27, 2024
Google ScholarTM
Check
Altmetric
Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.