HILIC-UPLC-FLR-ESI MS/MS

HILLIC-UPLC-FLR-ESI-MS/MS image

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.

Report

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)
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