This module is an extension of the Level 1 HILIC profiling work to gain more detailed information on the glycan structures and their relative proportions. This module provides HILIC-UHPLC-FLR profiles, ESI-MS/MS spectra, mass composition and sequence data matching glucose unit (GU) values, m/z and MS/MS sequencing data.
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 module, alongside Ludger positive and negative controls, and system suitability standards.
This module is suitable for:
- glycan assignment based on mass composition sequence data
- quality control – profile comparisons to monitor known structures
- monitoring batch to batch consistency
- comparability studies
In order to gain more detailed information (such as monosaccharide type and linkage) and increase confidence in the glycan structures assignments and their relative proportions Level 2 Exoglycosidase digestion followed by HILIC-UPLC is recommended.
Workflow for Level 2 HILIC-UPLC-FLR-ESI-MS/MS
Released and fluorescently labelled glycans (Level 1 HILIC profiling) are analysed by HILIC-UHPLC-FLR-ESI-MS/MS.
Final report contains:
- HILIC-UPLC profiles for system suitability standards and Ludger positive controls
- HILIC-UPLC profiles for client samples and buffer negative controls
- Glucose unit (GU) values
- Mass composition data matching GU, m/z and MS/MS sequencing data
- Proposed glycan structures and their relative proportions
- Summary of findings (e.g. G0:G1:G2; % high mannose; % fucosylation)