WAX Profiling

WAX-HPLC glycan analysis image

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.


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
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