Appliance

Identification of N-glycosylation sites in glycosylated proteins

Operation method

Biopharmaceutical Glycosylation Site Assay

Materials and Instruments

[Material] Biological medicine

Move

(1) Enzymatic cleavage of samples using trypsin or other appropriate proteases;(2) Separation of N-glycopeptides, O-glycopeptides and other peptides on a suitable column；

（3） Deglycosylation；

（4） Analyze the peptide profiles by MALDI-TOF MS, ESI-MS or MS/MS to find out the theoretical position of glycosylation；

（5） Determine the site of each N-O-glycogen by peptide mapping；

（6） Comparison of glycosylation sites in different batches of samples.

Caveat

After receiving the samples, the samples were first enzymatically digested to release the Fc domain and Fab of the antibody, and then the glycosylated fragments were isolated and the glycosylation sites were confirmed by MALDI TOF MS or nano LC-ESI-MS/MS.

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