This module provides MALDI-TOF-MS N- and/or O-glycan spectra, and m/z data for mass composition matching of the charged and neutral glycans. The analysis will be performed on a triplicate release of samples (typically 5-100 µg for each release), an equivalent amount (by volume) of sample buffer negative controls, alongside Ludger positive and negative controls and system suitability standards.
This module is suitable for:
- preliminary glycan assignment
- quality control – profile comparisons to monitor known structures
- monitoring batch to batch consistency
- comparability studies
Workflow for Level 1 MALDI profiling
N-glycans are released from glycoprotein by digestion with PNGAse F or PNGAse A.
O-glycans are released from glycoprotein by hydrazinolysis or Orela reagent.
Released glycans are permethylated, purified and analysed by MALDI-TOF-MS.
Permethylation is the process of derivatizing all the hydroxyl and N-acetyl groups by the addition of a methyl group. Permethylation also methyl esterifies the carboxy function on the sialic acid. Both these derivatizations lead to a stabilization of the sialic acids and to a significant enhancement in the MS analysis.
Final report contains:
- MALDI-TOF-MS spectra for system suitability standards and Ludger positive controls
- MALDI-TOF-MS spectra for client samples and buffer negative controls
- m/z data for mass composition matching of the charged and neutral glycans
- Relative proportions of detected peaks
- Summary of findings