Related Experiment Video
Updated: Aug 5, 2026
08:51
Profiling of Permethylated Mucin O-glycans Using Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry
Published on: June 20, 2025
Profiling O-Acetylation in Sialoglycans Using MALDI-TOF and LC-MS/MS with Methylamidation and Permethylation
Zhaoguan Wu1, Isabelle Sirois2
1CHU Sainte-Justine Research Center, Université de Montréal, Montreal, QC, Canada. zhaoguan.wu.hsj@ssss.gouv.qc.ca.
Methods in Molecular Biology (Clifton, N.J.)
|August 1, 2026
Summary
Analyzing O-acetylation patterns in N-sialoglycans is crucial for understanding diseases. This study presents a new workflow using methylamidation and permethylation for sensitive O-acetylation profiling in serum, aiding biomarker discovery.
Area of Science:
- Glycomics
- Biochemistry
- Analytical Chemistry
Background:
- O-acetylation of sialoglycans significantly impacts glycoprotein functions, influencing immune responses, cell signaling, and disease.
- N-acetylneuraminic acid (Neu5Ac) derivatives are key biomarkers for cancers and inflammatory diseases.
- Analyzing O-acetylation patterns is difficult due to the chemical lability of these modifications.
Purpose of the Study:
- To develop a robust workflow for analyzing O-acetylation patterns in serum/plasma-derived N-sialoglycans.
- To overcome the challenges posed by the chemical lability of O-acetylated glycans.
- To enable sensitive and reproducible profiling of sialylated glycan structures for biomarker discovery.
Main Methods:
- A novel workflow combining methylamidation and permethylation was developed.
- Porous graphitic carbon (PGC)-based purification was employed for sample cleanup.
- High-resolution MALDI-MS and LC-MS/MS were utilized for sensitive glycan analysis.
Main Results:
- The workflow demonstrated sensitive and reproducible profiling of O-acetylation patterns in N-sialoglycans.
- Successful analysis of O-acetylation in serum/plasma-derived samples was achieved.
- The method provides deeper insights into glycan modification dynamics.
Conclusions:
- This protocol offers a robust method for analyzing O-acetylation in N-sialoglycans.
- The developed workflow facilitates biomarker discovery and comparative glycomics studies.
- The approach enhances understanding of glycan modification in disease pathology.
