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Related Experiment Video

Updated: Sep 13, 2025

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Development and application of GlycanDIA workflow for glycomic analysis.

Yixuan Xie1,2,3, Xingyu Liu4, Li Yi5

  • 1State Key Laboratory of Genetic Engineering, Greater Bay Area Institute of Precision Medicine (Guangzhou), School of Life Sciences and Institutes of Biomedical Sciences, Fudan University, Shanghai, China. xieyixuan@ipm-gba.org.cn.

Nature Communications
|August 1, 2025
PubMed
Summary

We developed GlycanDIA, a sensitive method for identifying and quantifying glycans. This workflow reveals distinct glycan profiles on RNA and proteins, suggesting unique biological roles.

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Area of Science:

  • Glycomics and Mass Spectrometry
  • Cellular Biology and Disease Mechanisms

Background:

  • Glycans form the cell surface glycocalyx, regulating cellular processes.
  • Altered glycosylation patterns are implicated in disease initiation and progression.
  • Existing glycomic methods face limitations in sensitivity and precision for complex samples.

Purpose of the Study:

  • To introduce GlycanDIA, a novel data-independent acquisition (DIA)-based glycomic workflow.
  • To develop GlycanDIA Finder, a generic search engine for confident glycan identification from DIA data.
  • To profile and compare N-glycans on RNA and proteins, including low-abundance species.

Main Methods:

  • Implementation of a DIA-based glycomic workflow combining higher energy collisional dissociation (HCD)-MS/MS and staggered windows.
  • Development of the GlycanDIA Finder search engine with iterative decoy searching for enhanced glycan identification.
  • Application of the workflow to distinguish glycan composition and isomers (N-glycans, O-glycans, HMOs) and profile glycoRNA.

Main Results:

  • GlycanDIA demonstrates high sensitivity and precision in glycan identification and quantification.
  • The workflow successfully differentiates glycan isomers and identifies low-abundant modified glycans.
  • Analysis of glycoRNA revealed distinct N-glycan abundance compared to protein-glycans, with tissue-specific variations.

Conclusions:

  • GlycanDIA offers a significant advancement for sensitive and precise glycomic analysis.
  • The developed workflow enables comprehensive profiling of N-glycans on RNA molecules.
  • Distinct glycoRNA profiles suggest unique functional roles for RNA-glycans in biological processes and tissues.