N-Glycosylation site analysis is crucial for understanding the functions of glycans. Yaohai Bio-Pharma uses Liquid Chromatography-Mass Spectrometry (LC-MS) after enzymatic cleavage for N-Glycosylation site.
We have been involved in the Protein Structural Characterization of various large molecules, including Recombinant Subunit Vaccines, Nanobodies/VHHs/ Single-domain Antibodies (sdAbs), Antibody Fragments, Hormones/Peptides, Cytokines, Growth Factors (GF), Enzymes, Collagens, etc.
Regulatory Requirements for N-Glycosylation Site
According to ICH Q6B guideline, “for glycoproteins, the carbohydrate content (neutral sugars, amino sugars, and sialic acids) is determined. Furthermore, the structure of the carbohydrate chains, the oligosaccharide pattern (antennary profile), and the glycosylation site(s) of the polypeptide chain are analyzed to the best extent possible.
Analytical Method
| Analysis |
Methods |
| N-glycosylation site |
Liquid Chromatography-Mass Spectrometry (LC-MS) after enzymatic cleavage |
Procedures
1. Enzyme digestion of the protein using trypsin or other protease
2. Separate fragment glycopeptides
3. Confirm the N-glycosylation site by LC-MS
4. Analyze data
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