CD BioGlyco has abundant experience in Antibody Deglycosylation. We expect to become customers' research assistants in the hydrolysis of complex-type N-glycan.

A Key Tool to Glycoengineer Immunotherapeutic IgG: EndoS2 Glycosynthase Mutants

N-glycosylation is one of the most abundant posttranslational modifications of proteins, essential for many physiological processes, including protein folding, protein stability, oligomerization and aggregation, and molecular recognition events. The ability to manipulate protein N-glycosylation is critical not only to our fundamental understanding of biology but also for the development of new drugs for a wide range of human diseases.

EndoS2 glycosynthase mutants from S. pyogenes are key tools to glycoengineer immunotherapeutic IgG monoclonal antibodies. EndoS is a bacterial endo-β-N-acetylglucosaminidase that specifically catalyzes the hydrolysis of the β-1, 4 linkage between the first two N-acetylglucosamine residues of the biantennary complex-type N-linked glycans of IgG Fc regions. It is used for the chemoenzymatic synthesis of homogeneously glycosylated antibodies with improved therapeutic properties. Because of the broad N-glycan specificity of EndoS2, this enzyme can hydrolyze, and as a glycosynthase mutant to generate a more diverse set of N-glycans on antibodies.

Fig.1 Schematic representation of glycan hydrolysis of N-linked glycans. (Garbe & Collin, 2012)

Hydrolysis of Complex-Type N-Glycans

CD BioGlyco provides an endoglycosidase that specifically hydrolyzes N-linked glycans in the Fc region of native IgG. The endoglycosidase has excellent specificity and only hydrolyzes glycans from the IgG Fc domain when other glycosylated proteins are present in the system.

The endoglycosidase can be used to rapidly deglycosylate antibodies to reduce sample complexity, or to inactivate Fc-mediated effector functions. However, it has limited activity against high mannose and hybrid glycans. Please click here if you have needs for Hydrolysis of All Fc-Glycans in IgGs.

Applications

• Genetic modification of multiple expression systems to produce glycoconjugation-based site-specific antibody-drug conjugates
• Recombination of N-glycans to obtain uniform glycoforms or chemically modified glycans that can control or modify antibody properties
• High homogeneity and precise control of sugar morphology by in vitro chemoenzymatic remodeling

Advantages of Us

• An advanced Glycoengineering Platform
• A variety of IgGs from many different species, including all human IgG subclasses, IgGs from mice, rabbits, rats, monkeys, sheep, goats, cows, and horses
• Comprehensive and reliable after-sales service

CD BioGlyco constantly updates the technology platform to provide customers with comprehensive and high-quality services for hydrolysis of complex-type N-glycan. We will continue to raise the standard to meet customers' research needs. To learn more, please don't hesitate to contact us to learn how we can help you with your project.

References:

1. Garbe, J.; Collin, M. Bacterial hydrolysis of host glycoproteins–powerful protein modification and efficient nutrient acquisition. Journal of innate immunity. 2012, 4(2): 121-131.
2. Trastoy, B.; et al. GH18 endo-β-N-acetylglucosaminidases use distinct mechanisms to process hybrid-type N-linked glycans. Journal of Biological Chemistry. 2021, 297(2).
3. Trastoy, B.; et al. Structural basis for the recognition of complex-type N-glycans by Endoglycosidase S. Nature communications. 2018, 9(1): 1-11.