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Updated: Mar 8, 2026
Spatial Molecular Imaging of the Glycome Using Mass Spectrometry
Published on: November 28, 2025
MALDI Mass Spectrometry Imaging of N-Linked Glycans in Cancer Tissues
R R Drake1, T W Powers1, E E Jones1
1Medical University of South Carolina, Charleston, SC, United States.
Abstract:
Glycosylated proteins account for a majority of the posttranslation modifications of cell surface, secreted, and circulating proteins. Within the tumor microenvironment, the presence of immune cells, extracellular matrix proteins, cell surface receptors, and interactions between stroma and tumor cells are all processes mediated by glycan binding and recognition reactions. Changes in glycosylation during tumorigenesis are well documented to occur and affect all of these associated adhesion and regulatory functions. A MALDI imaging mass spectrometry (MALDI-IMS) workflow for profiling N-linked glycan distributions in fresh/frozen tissues and formalin-fixed paraffin-embedded tissues has recently been developed. The key to the approach is the application of a molecular coating of peptide-N-glycosidase to tissues, an enzyme that cleaves asparagine-linked glycans from their protein carrier. The released N-linked glycans can then be analyzed by MALDI-IMS directly on tissue. Generally 40 or more individual glycan structures are routinely detected, and when combined with histopathology localizations, tumor-specific glycans are readily grouped relative to nontumor regions and other structural features. This technique is a recent development and new approach in glycobiology and mass spectrometry imaging research methodology; thus, potential uses such as tumor-specific glycan biomarker panels and other applications are discussed.
Insights
This study introduces a new method using MALDI imaging mass spectrometry to map N-linked glycans directly on tissue. This technique aids in identifying tumor-specific glycans for potential biomarker development.
Area of Science:
- Glycobiology
- Mass Spectrometry Imaging
- Cancer Research
Background:
- Glycosylated proteins are crucial for cell surface functions and are altered during tumorigenesis.
- Tumor microenvironment interactions rely heavily on glycan binding and recognition.
- Post-translational modifications, particularly glycosylation, play a significant role in cancer progression.
Purpose of the Study:
- To develop and present a novel workflow for profiling N-linked glycan distributions directly within tissue samples.
- To demonstrate the capability of MALDI imaging mass spectrometry in analyzing glycans in both fresh/frozen and FFPE tissues.
- To explore the potential of this technique for identifying tumor-specific glycans and developing biomarker panels.
Main Methods:
- A workflow utilizing peptide-N-glycosidase to cleave N-linked glycans from proteins directly on tissue sections.
- Analysis of released N-linked glycans using MALDI imaging mass spectrometry (MALDI-IMS).
- Integration of glycan profiling with histopathology for spatial localization and comparison between tumor and non-tumor regions.
Main Results:
- Routine detection of 40 or more individual glycan structures per sample.
- Successful profiling of N-linked glycans in fresh/frozen and formalin-fixed paraffin-embedded (FFPE) tissues.
- Ability to localize tumor-specific glycans relative to histopathological features.
Conclusions:
- The developed MALDI-IMS workflow offers a powerful new approach for glycobiology and mass spectrometry imaging research.
- This technique facilitates the identification of spatially defined, tumor-specific glycans.
- Potential applications include the development of novel glycan-based biomarker panels for cancer diagnostics and prognostics.
