CD BioGlyco has an advanced Glycoengineering Platform and expertise in glycoscience. In addition, our professional research teams allow us to provide you with high-quality services for the hydrolysis of all Fc-glycans.

Enzymatic Removal of the Fc-glycan in the Glycoengineering of Therapeutic Antibodies

Glycoengineering of therapeutic antibodies has attracted considerable attention. Recently, scientists have been intrigued by two enzymes from the human pathogen, EndoS and EndoS2 from Streptococcus pyogenes. Initially, these enzymes were only thought of as bacterial immune evasion factors that eliminate the effector function of immunoglobulin G (IgG) by hydrolyzing N-linked glycans on antibodies. IgG carries a complex N-linked glycan on each C_H2 domain, an oligosaccharide essential for the structure of the Fc region and interaction with Fc receptors. Removal of Fc glycans by EndoS resulted in a significant reduction in IgG binding of the Fc region to Fcγ receptors. Both EndoS and EndoS2 catalyze the hydrolysis of the β-1,4 linkage between two N-acetylglucosamine (GlcNAcs) in the N-linked glycan core of human IgG.

The effect of the IgG glycans on antibody functions has gained major attention in the growing field of monoclonal therapeutic antibodies. To enhance the efficacy of the therapeutic antibodies, the focus is turning toward modifying the Fc part of the antibody to specifically interact with selected Fcγ receptors. The therapeutic antibody is used to elicit the desired immune response in this way, thereby increasing the serum half-life of the antibody. Interactions can also be engineered by mutagenizing amino acid residues in the hinge region involved in binding and designing oligosaccharides linked to Asn297 on the IgG heavy chain. The increasing availability of biotechnology tools to control, study and direct the glycosylation of IgG facilitates the development of therapeutic antibodies with preselected glycoforms. The specificity of IgG makes the developed enzyme useful as a tool for glycoengineering.

Unlocking the True Potential of Your Antibodies: Precision Hydrolysis of All Fc-Glycans

It was found that EndoS2 hydrolyzed high mannose and hybrid structures on therapeutic antibodies to a greater extent than EndoS. In combination with the IdeS protease, CD BioGlyco provides a rapid EndoS-EndoS2-based assay for quantifying high mannose and hybrid Fc-glycosylation on monoclonal antibodies.

CD BioGlyco offers an IgG-specific endoglycosidase that hydrolyzes all glycoforms present at Fc glycosylation sites. The endoglycosidase specifically hydrolyzes glycans at the IgG Fc glycosylation site without affecting the core GlcNAc. In addition, the endoglycosidase is capable of deglycosylating IgG isoforms from multiple species. Please click here if you have needs for hydrolysis of complex type N-glycans: Hydrolysis of Complex-Type N-Glycans.

Project Proposal

Our experienced team of scientists will work with you to understand your specific project goals, sample types, and expected results. We will use this information to design a customized deglycosylation strategy tailored to your unique needs.

Sample Preparation

Upon receipt, samples undergo rigorous quality control to assess purity, concentration, and integrity. This step ensures high-quality starting material, which is critical for successful deglycosylation and accurate downstream analysis.

Enzymatic Deglycosylation

Using our optimized enzymatic cocktail, your antibody sample undergoes a controlled and efficient deglycosylation reaction. This step is carefully monitored to ensure complete hydrolysis of all Fc glycans while maintaining the stability of the antibody structure.

Purification

Following the deglycosylation reaction, the sample is treated to remove the enzyme, released glycans, and any other reaction byproducts. This purification step yields a pure, fully deglycosylated antibody preparation ready for subsequent experiments.

Data Reporting and Analysis

We provide a comprehensive report covering the entire process, including initial quality control data, deglycosylation efficiency, and final sample purity. We also provide a range of analytical services to confirm the success of deglycosylation, ensuring you have confidence in your results.

Workflow

Publication Data

Applications

• Achieving uniform glycosylation is crucial for understanding a therapeutic's mechanism of action and ensuring consistent potency and safety.
• The removal of complex glycans simplifies analysis, enabling high-resolution structural studies via X-ray crystallography, nuclear magnetic resonance (NMR), and mass spectrometry (MS).
• Researchers use deglycosylated antibodies to investigate the specific roles of glycans in antibody-antigen binding, Fc receptor interactions, and complement activation.

Advantages

• Our specialized enzymatic cocktail ensures the uniform and complete removal of all N-linked glycans from the Fc region, providing a homogeneous antibody population for your experiments.
• Unlike harsh chemical methods, our gentle enzymatic approach maintains the structural and functional integrity of the antibody, preserving its native conformation for reliable results.
• Our optimized workflow delivers consistent and reproducible outcomes, critical for a variety of applications, from basic research to preclinical development.

Frequently Asked Questions

With extensive experience, rich industry knowledge and advanced Antibody Deglycosylation technologies, CD BioGlyco is exactly competent in the hydrolysis of all Fc-glycans. We are confident in accomplishing even the most challenging engineering project with a quick turnaround time. To learn more, please don't hesitate to contact us to learn how we can help you with your project.

Associated Services

Quick Links

Resources