The glycan structure is a critical attribute of your protein!
We offer determination of the number, position and type of glycosylation in your drug substance protein. The analysis can be used for comparing different batches or originator versus biosimilar.
Glycosylation of proteins is one of the most common post translation modifications in biopharmaceuticals. Glyco-proteins contain complex sugar moieties whose presence, absence, sites of attachment, and relative abundance can have a major impact on the efficacy and stability of the drug substance.
Glycosylation analysis can be done at three different levels and you can combine one or more to get the full glycan characterization:
Part 1: Analysis on intact protein
- Exact molecular weight of glycoprotein determined by UV-LC ESI-MS
- Exact molecular weight of deglycosylated protein determined by UV-LC ESI-MS
Part 2: Glyco-peptide mapping
- Glycoprotein is digested with one of more proteases and analyzed by nano LC-MS/MS
- Searched against drug substance sequence and glycostructure database
Part 3: Released N-glycan analysis and quantitation
- PNGase F treatment to release N-linked glycans
- Fluorescent labelling using RapiFluor-MS labelling kit
- Separation of glycans by HILIC chromatography, detection by fluorescence HPLC and mass spectrometry
- Glycan profile and type by FLD-trace and glycan mass
You can also send us a request so we can adjust the analysis to meet your specific needs