Scientific Publications
Profiling of permethylated mucin O-glycans using Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
Journal of Visualised Experiments
Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) is a powerful tool for the profiling of mucin glycans that offers high sensitivity and throughput. Here, we provide a detailed protocol for the MALDI-TOF MS analysis of O-linked glycans from mucins, encompassing glycan release, desalting, permethylation, and data acquisition and analysis. We show that desalting based on ion exchange chromatography leads to improved recovery of larger glycans and better quality of the produced spectra compared to solid-phase extraction using porous graphitiz[NJ1] ed carbon (PGC). Additionally, our work suggests that a longer permethylation incubation than currently employed might be necessary for efficient modification of sialylated structures. We also demonstrate the importance of fragmentation data for the accurate identification of glycan composition and structure and discuss the inherent benefits and limitations that MALDI-TOF presents in the analysis of mucin O- glycans.
Journal of Visualised Experiments
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