Glycosylation Sites (N-Glycosylation, O-Glycosylation) Detection for Antibody Drugs

Antibody drugs are a class of drugs that use antibodies to specifically recognize and bind to target molecules. Glycosylation is a critical type of post-translational modification that can affect the stability, biological activity, and immune response of antibody drugs. Glycosylation typically occurs at the N (asparagine) and O (serine or threonine) sites of the antibody. N-glycosylation often occurs in a specific sequence between asparagine and serine (N-X-S/T), while O-glycosylation primarily occurs on serine and threonine residues. Detection and analysis of these glycosylation sites are crucial for ensuring the quality and efficacy of antibody drugs.

 

There are many methods for analyzing the glycosylation sites of antibody drugs. Mass spectrometry is a common method that can provide precise information about the composition and linkage of the glycan chains. This usually involves enzymatically digesting the antibody to produce peptides with glycosylation sites, which can then be determined by mass spectrometry to determine the structure and location of the glycan chains. Another common technique is liquid chromatography, which can quantitatively analyze the glycan chains. By using specific enzymes to release N-glycans and O-glycans, followed by fluorescent labeling and liquid chromatography analysis, quantitative information about the glycan chains can be obtained. In addition, there are some more complex techniques, such as nuclear magnetic resonance and X-ray crystallography, that can be used to study glycosylation structures. By using these methods, we can gain a comprehensive understanding of the glycosylation of antibody drugs to optimize their production process and enhance the effects of the drugs.

 

MtoZ Biolabs provides drug quality research services that comply with global regulatory laws and regulations for customers. We have two mass spectrometry analysis technologies, MALDI TOF MS and nano LC-ESI-MS/MS, to provide you with efficient and accurate antibody drug glycosylation identification services. After receiving the sample, we first digest the sample to release the Fc domain and Fab of the antibody, then separate the glycosylated fragments, and confirm the glycosylation site through MALDI TOF MS or nano LC-ESI-MS/MS.