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Related Experiment Video

Updated: May 13, 2026

Glycomics-Guided Glycoproteomics Facilitates Comprehensive Profiling of the Glycoproteome in Complex Tumor Microenvironments
10:59

Glycomics-Guided Glycoproteomics Facilitates Comprehensive Profiling of the Glycoproteome in Complex Tumor Microenvironments

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Glycomic profiling of tissue sections by LC-MS.

Yunli Hu1, Shiyue Zhou, Sarah I Khalil

  • 1Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas 79409, United States.

Analytical Chemistry
|February 27, 2013
PubMed
Summary

A new glycan sample preparation protocol minimizes steps, enabling sensitive N-glycan analysis from small tissue samples. This method provides detailed spatial information for glycomic research.

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Area of Science:

  • Glycomics
  • Mass Spectrometry
  • Biochemistry

Background:

  • Traditional glycan analysis requires large samples due to sample loss during multi-step preparation.
  • Existing methods lack detailed molecular spatial information.
  • Proper storage and shipping of samples are critical and challenging.

Purpose of the Study:

  • To develop a novel, streamlined glycan sample preparation protocol.
  • To enable sensitive N-glycan analysis from minimal sample amounts.
  • To facilitate the assessment of glycan spatial distribution in tissues.

Main Methods:

  • On-surface enzymatic digestion of tissue sections and spotted samples to release N-glycans.
  • Reduction and permethylation of released glycans.

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  • Online purification coupled with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis.

    • Main Results:

    • Detection of permethylated N-glycans from as little as 10 ng of RNase B.
    • Identification of 66 N-glycans from a 0.1-μL aliquot of human blood serum (HBS).
    • Detection of 43 N-glycans from nude mouse brain tissue, comparable to larger-scale studies, using submicrogram samples.

    Conclusions:

    • The developed protocol offers high sensitivity and reproducibility for N-glycan profiling.
    • This method allows for the analysis of glycans from significantly smaller sample sizes compared to routine methods.
    • The on-tissue N-glycan profiling approach is suitable for assessing glycan spatial distribution and can be correlated with imaging techniques.