Exoglycosidase HILIC Analysis

exoglycosidase digestion followed by hilic analysis image

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)
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