Issue 4, 2015

PNGase F-mediated incorporation of 18O into glycans for relative glycan quantitation

Abstract

PNGase F-catalyzed glycosylation site 18O-labeling is a widely used method for glycoprotein quantitation owing to its efficiency and simplicity. However, PNGase F-catalyzed glycan 18O-labeling, which offers advantages for glycomics, has not yet been developed. In this study, PNGase F-mediated incorporation of 18O into glycans during the N-glycan release from glycoproteins by PNGase F was finally realized, named as PCGOL (PNGase F-catalyzed glycan 18O-labeling), which offers a potential strategy for relative glycan quantitation. This new method showed good linearity and high reproducibility within at least 2 orders of magnitude in the dynamic range. Furthermore, PCGOL combined with our previously developed TOSIL method (tandem 18O stable isotope labeling for N-glycoproteome quantitation) can be used for comprehensive N-glycosylation quantification, achieving simultaneous quantification of glycans, glycopeptides and glycoproteins in a single workflow, which was also used to analyze glycosylation changes in immunoglobulin G (IgG) associated with hepatocellular carcinoma in the present work.

Graphical abstract: PNGase F-mediated incorporation of 18O into glycans for relative glycan quantitation

Supplementary files

Article information

Article type
Paper
Submitted
11 Nov 2014
Accepted
03 Dec 2014
First published
03 Dec 2014

Analyst, 2015,140, 1082-1089

Author version available

PNGase F-mediated incorporation of 18O into glycans for relative glycan quantitation

W. Zhang, W. Cao, J. Huang, H. Wang, J. Wang, C. Xie and P. Yang, Analyst, 2015, 140, 1082 DOI: 10.1039/C4AN02073A

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