This module provides WAX-HPLC charge profiles with the relative proportion of the mono-, di-, tri- and tetra-sialylated glycan structures. The analysis will be performed on single or triplicate releases of samples (typically 5-100 µg for each release), an equivalent amount (by volume) of sample buffer negative control, and alongside Ludger’s system suitability standards, positive and negative controls.
This module is suitable for:
- sialylated samples
- quality control – profile comparisons to monitor charge distribution
- monitoring batch to batch consistency
- comparability studies
In order to gain more detailed information on the glycan structures and their relative proportions Level 2 Detailed Characterisation is required.
Workflow for Level 1 WAX 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 fluorescently labelled with 2-AB purified and analysed on a LudgerSep-C3 weak anion exchange (WAX) column.
WAX-HPLC provides charge profiles for profile comparison between batches where glycan separation is based on glycan charge.
Report
Final report contains:
- WAX-HPLC profiles for system suitability standards and Ludger positive controls
- WAX-HPLC profiles for client samples and buffer negative controls
- Relative proportions of the mono-, di-, tri- and tetra-sialylated glycan structures
