Glycosylation analysis
- Exact molecular weight of glycoproteins
- Peptide map of glycopeptides
- Analysis of released N-glycans
Determine the glycan structure and heterogeneity of protein or antibody
Glycosylation of proteins is one of the most common post-translation modifications (PTMs) and also a critical quality attribute of biopharmaceutical products. Glycoproteins contain complex sugar moieties whose presence, absence, sites of attachment, and relative abundance can significan...
Determine the glycan structure and heterogeneity of protein or antibody
Glycosylation of proteins is one of the most common post-translation modifications (PTMs) and also a critical quality attribute of biopharmaceutical products. Glycoproteins contain complex sugar moieties whose presence, absence, sites of attachment, and relative abundance can significantly impact the efficacy, stability, and activity of proteins and monoclonal antibodies (mAbs). Hence, it is essential to determine the glycan structure and heterogeneity through antibody glycosylation analysis.
A complete protein glycosylation analysis includes the number, position, and type of glycosylation in your protein or mAb. The study is beneficial for comparing different batches, originator versus biosimilar, and process development samples.
Which glycosylation analysis services should you choose?
Protein glycosylation analysis can be done at different levels. It can also be combined to achieve a complete glycan characterization. Below you can see examples of our services:
Service 1: Separation and exact molecular weight of protein/ antibody glycoforms
- Firstly, determination of the exact molecular weight of glycoprotein by LC-MS (HILIC separation and online LC-MS).
- Then, determination of the exact molecular weight of deglycosylated protein by LC-MS (HILIC separation and online LC-MS).
Service 2: Separation and exact molecular weight of mAb sub-units
- IdeS digestion and reduction to sub-units.
- HILIC separation of LC, Fd, and single chain Fc (scFc) glycoforms.
- Online LC-MS to determine the mass of each sub-unit.
Service 3: Glycopeptide map
- Glycoprotein digest with one of more proteases (trypsin or Lys-C) and analyzed by LC-MS/MS.
- Identification of glycopeptides by searching against drug substance sequence and glycostructure database.
Service 4: Released N-glycan analysis
- PNGase F treatment to release N-linked glycans.
- Fluorescent labeling using Waters RapiFluor Glycan labeling kit.
- Separation of glycans by HILIC chromatography, detection by fluorescence HPLC, and mass spectrometry.
- Identification of both glycan profile and type by FLD-trace and glycan mass.
Please send us a request so we can adjust the glycosylation analysis to meet your specific needs.
You can also take a look at our other protein characterization services.