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

Analyst. 2015 Feb 21;140(4):1082-9. doi: 10.1039/c4an02073a.


PNGase F-catalyzed glycosylation site (18)O-labeling is a widely used method for glycoprotein quantitation owing to its efficiency and simplicity. However, PNGase F-catalyzed glycan (18)O-labeling, which offers advantages for glycomics, has not yet been developed. In this study, PNGase F-mediated incorporation of (18)O into glycans during the N-glycan release from glycoproteins by PNGase F was finally realized, named as PCGOL (PNGase F-catalyzed glycan (18)O-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 (18)O 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carbohydrate Sequence
  • Carcinoma, Hepatocellular / metabolism
  • Glycomics / methods
  • Glycopeptides / chemistry
  • Glycopeptides / metabolism
  • Glycoproteins / chemistry*
  • Glycoproteins / metabolism
  • Glycosylation
  • Humans
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / metabolism
  • Liver Neoplasms / metabolism
  • Mass Spectrometry / methods
  • Molecular Sequence Data
  • Oxygen Isotopes / analysis
  • Oxygen Isotopes / metabolism*
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism*
  • Polysaccharides / analysis*
  • Polysaccharides / metabolism
  • Reproducibility of Results


  • Glycopeptides
  • Glycoproteins
  • Immunoglobulin G
  • Oxygen Isotopes
  • Polysaccharides
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase