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Related Experiment Video

Updated: Dec 22, 2025

Profiling of Permethylated Mucin O-glycans Using Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry
08:51

Profiling of Permethylated Mucin O-glycans Using Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry

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Simplifying Glycan Profiling through a High-Throughput Micropermethylation Strategy.

Asif Shajahan1, Nitin T Supekar1, Digantkumar Chapla1

  • 1Complex Carbohydrate Research Center, The University of Georgia, Athens, GA, USA.

SLAS Technology
|May 5, 2020
PubMed
Summary
This summary is machine-generated.

We developed a microscale permethylation strategy for high-throughput glycan analysis. This method enables efficient structural characterization of glycans from diverse biological sources using automated mass spectrometry.

Keywords:
automated ESI-MSglycan branchingglycomicshigh-throughput glycan analysispermethylation

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Glycomics

Background:

  • Glycoproteins are crucial biomolecules with complex functions, but their analysis is challenging due to heterogeneity and low abundance.
  • Glycosylation profiles critically influence the properties of therapeutic glycoproteins like antibodies and vaccines.
  • Accurate glycan characterization is essential for quality control and development of glycoprotein-based therapeutics.

Purpose of the Study:

  • To present a novel microscale permethylation strategy for high-throughput glycan analysis.
  • To demonstrate the utility of this method for structural characterization of glycans from various biological samples.
  • To enable automated, in-depth analysis of N- and O-glycans.

Main Methods:

  • Development of a microscale permethylation strategy for efficient glycan derivatization.
  • High-throughput sample processing using 96-well plates and microcentrifuge tubes.
  • Automated tandem electrospray ionization-mass spectrometry (ESI-MSn) platform for structural analysis.
  • C18 tip-based cleanup for simplified glycan isolation.

Main Results:

  • Successful implementation of microscale permethylation for nanogram-level glycan processing.
  • Demonstrated high-throughput structural analysis of glycans from human plasma, mammalian cells, and purified glycoproteins.
  • Automated multistage tandem MS acquisition enabled in-depth characterization, including sialic acid linkages and bisecting N-acetylglucosamine (GlcNAc).

Conclusions:

  • The micropermethylation strategy significantly enhances throughput for glycan analysis.
  • This automated workflow facilitates the structural characterization of diverse glycans.
  • The approach is valuable for high-throughput screening of glycosylation in clinical glycan biomarkers and glycoprotein therapeutics.