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Updated: Dec 10, 2025

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
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Quantitative N-glycoproteomics using stable isotopic diethyl labeling.

Yue Wang1, Kaijie Xiao1, Zhixin Tian1

  • 1School of Chemical Science & Engineering, Shanghai Key Laboratory of Chemical Assessment and Sustainability, Tongji University, Shanghai, 200092, China.

Talanta
|September 5, 2020
PubMed
Summary

Stable isotopic diethyl labeling (SIDE) enables accurate, large-scale quantification of intact N-glycopeptides. This method reveals differentially expressed N-glycopeptides (DEGPs) crucial for understanding cancer and other diseases.

Keywords:
Diethyl labelingDifferentially expressionIntact N-glycopeptidesN-glycoproteomicsQuantitation

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Last Updated: Dec 10, 2025

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Area of Science:

  • Biochemistry
  • Proteomics
  • Glycobiology

Background:

  • Protein N-glycosylation is vital in cancer and other diseases.
  • Accurate qualitative and quantitative analysis of N-glycosylation is essential for structural and functional studies.
  • Quantifying differentially expressed intact N-glycopeptides (DEGPs) at the molecular level is critical.

Purpose of the Study:

  • To develop and apply a novel method for relative quantitation of intact N-glycopeptides.
  • To validate the method's accuracy and dynamic range.
  • To investigate aberrant N-glycosylation in gastric cancer.

Main Methods:

  • Stable isotopic diethyl labeling (SIDE) of amino groups on N-glycopeptides using isotopically labeled acetaldehyde (acetaldehyde-13C or acetaldehyde-C) and sodium cyanoborohydride.
  • Diethylation of both N-terminal and lysine residues.
  • Application of SIDE in comparative N-glycoproteomics.

Main Results:

  • Achieved a linear quantitation dynamic range up to 50-fold (R2 = 0.9985) using standard glycoprotein ribonuclease B.
  • Identified 644 DEGPs with a fold change ≥1.5 and p < 0.05 in a comparative study of gastric cancer versus adjacent tissues.
  • Demonstrated the method's accuracy and broad dynamic range for intact N-glycopeptide analysis.

Conclusions:

  • Stable isotopic diethyl labeling (SIDE) is an effective technique for the relative quantitation of intact N-glycopeptides.
  • SIDE offers high accuracy and a large dynamic range, suitable for quantitative N-glycoproteomics.
  • This method can be broadly applied to study aberrant glycosylation in various pathological conditions, including cancer.