This module provides HILIC-UPLC N- and/or O-glycan profiles before and after specific exoglycosidase digestions together with glycan structures and their relative quantitation. The analysis will be performed on single or triplicate N- and/or O-glycan samples, released and fluorescently labelled under the Level 1 HILIC profiling, alongside Ludger positive and negative controls and system suitability standards.
This module is suitable for:
- glycan assignment (monosaccharide type, order, linkage)
- quality control – profile comparisons to monitor known structures
- monitoring batch to batch consistency
- comparability studies
In order to gain more detailed information and increase confidence in the glycan structures assignment and their relative proportions additional Level 2 HILIC-UPLC-FLR-ESI-MS/MS is recommended.
Workflow for Level 2 Exoglycosidase digestion followed by HILIC-UPLC
Released and fluorescently labelled glycans (Level 1 HILIC profiling) are digested with a series or a mixture of specific exoglycosidases purified and analysed by HILIC-HPLC or HILIC-UHPLC.
Final report contains:
- HILIC-UPLC profiles for system suitability standards and Ludger positive controls before and after exoglycosidase digestions
- HILIC-UPLC profiles for client samples and buffer negative controls before and after exoglycosidase digestions
- Glucose unit (GU) values
- Glycan structures (including monosaccharide type, order and linkage) and their relative proportions
- Summary of findings (e.g. G0:G1:G2; % high mannose; % fucosylation; % Galα1-3Gal)