Antibody Deglycosylation Service
At present, antibody deglycosylation is being extensively studied. Understanding the function of glycosaminoglycans can help us deeply study immunotherapy. CD BioGlyco has developed advanced Antibody Glycoengineering technologies and analysis tools to help customers study antibody deglycosylation systematically.
Deglycosylated Antibodies for Therapeutics
Immunotherapy is one of the most encouraging strategies for cancer treatment, and the most common immunotherapy strategy involves the interruption of the interaction between immune checkpoints expressed on tumors and immune cells, which blocks the immune escape of tumor cells to some extent. The structure and function of monoclonal antibodies have become key research points in tumor therapy.
If a lower propensity to activate inflammatory cascades is desirable deglycosylated antibodies may be good candidates for therapeutics because the removal of glycan reduces the IgG interactions with Fc receptors. Deglycosylated antibodies have been of interest to treat autoimmune disorders and to suppress immune complex-mediated inflammation in a mouse arthritis model by disrupting Fc-Fc interactions while maintaining intact antigen-antibody binding and complement binding.
Antibody deglycosylation has been studied by EndoS (endo-β-N-acetylglucosaminidase) hydrolysis. EndoS is capable of cleaving glycans in the crystallizable fragment (Fc) domain of IgG molecules. This digestion significantly reduces the binding of immune system cell receptors to IgG.
Unlocking the Full Potential of Biotherapeutics with Antibody Deglycosylation
Deglycosylation of antibodies can significantly alter their intrinsic properties and stability, creating challenges for downstream process development. CD BioGlyco provides a fast and efficient technique to remove conserved glycans to eliminate the function of Fc in therapeutic monoclonal antibodies. This facilitates the study of the action mode of IgG.
Utilizing broad-spectrum enzymes, we ensure the hydrolysis of complex, high-mannose, and hybrid type N-glycans. This process yields a fully deglycosylated antibody, which is essential for studying the protein properties.
Hydrolysis of Complex Type N-Glycans
By employing specific endoglycosidases that selectively cleave these glycans, we simplify the glycoform profile of the antibody. This is particularly valuable for improving the homogeneity of a biotherapeutic.
Project Design
Our specialists work closely with you to understand your research goals, sample characteristics, and specific deglycosylation requirements. This ensures the selection of the most appropriate enzymes and analytical methods tailored to your project.
Sample Preparation
Upon receipt, your antibody samples undergo a rigorous quality control check to assess purity and integrity. Samples are then prepared for enzymatic treatment, including buffer exchange and optimization to ensure ideal conditions for glycosidase activity.
Controlled Enzymatic Deglycosylation
Samples are incubated with the selected glycosidases under precisely controlled conditions (temperature, pH, and incubation time). Our optimized protocols ensure complete deglycosylation without compromising the antibody's protein backbone.
Purification and Separation
After enzymatic treatment, the deglycosylated antibody is purified to remove the enzymes and any other residual contaminants. We employ advanced chromatography techniques to isolate the final product, ensuring a highly pure and homogeneous preparation.
Post-Deglycosylation Analysis
The deglycosylated antibody is subjected to a battery of analytical tests to verify the successful and complete removal of glycans, confirming the resulting mass and purity of the protein.
Workflow
Publication Data
Journal: Nature Communications
IF: 14.7
Published: 2023
Results: Streptococcal enzymes EndoS and EndoS2 mediate antibody-specific deglycosylation to enable bacterial immune evasion by disabling IgG function. They selectively hydrolyze the conserved N-glycan at IgG Fc's Asn297 via a dual mechanism, while its glycosidase domain targets the glycan. Cryo-EM reveals a 1:1 "V"-shaped complex orienting the active site for glycan access. Nuclear magnetic resonance (NMR) shows higher affinity for IgG Fc over free glycans, driven by protein-protein interactions enhancing binding on-rates. This targeted mechanism, via constrained glycan sites and protein anchoring, ensures specific IgG targeting, supporting immune evasion and guiding therapeutic engineering.
Fig.1 EndoS, SpeB, and IdeS act on IgG antibodies. (Trastoy, et al., 2023)
Applications
- Deglycosylation reduces IgG complement-dependent and antibody-dependent cell-mediated cytotoxicity with a reduction in antigen binding, giving it therapeutic potential
- Deglycosylation results in a more flexible Fc structure within IgG1, allowing a greater understanding of antibody structure
- Deglycosylated IgG has a dominant suppressive effect on inflammation
- Simplify the complex and tedious process of analyzing glycans
Advantages of Us
- Many years of experience in antibody deglycosylation
- Different experimental programs can be selected according to every special need
- A comprehensive and advanced Glycoengineering Platform
- Experienced and well-trained technicians
- Customized service to ensure your flexible choice in an efficient manner
Frequently Asked Questions
Our team of scientists has many years of experience in antibody deglycosylation. With our advanced Glycoengineering Platform, CD BioGlyco provides customers with strong support for antibody deglycosylation. For more detailed information and quote, please feel free to contact us for deep communication.
Associated Services
To help you achieve your overall research and development goals, CD BioGlyco offers a portfolio of complementary services that are used in conjunction with our antibody deglycosylation service.
- Glycomic Characterization: Understand the full glycan profile of your antibody before and after deglycosylation using our advanced glycan sequencing and profiling services.
- Antibody Glycoengineering: Modify the glycosylation of your antibody to enhance its function, stability, or half-life.
- Antibody-Drug Conjugate (ADC) Development: Leverage our expertise in site-specific conjugation to create next-generation ADCs with improved homogeneity and efficacy.
Reference
- Trastoy, B.; et al. Mechanism of antibody-specific deglycosylation and immune evasion by Streptococcal IgG-specific endoglycosidases. Nature Communications. 2023, 14(1): 1705. (Open Access)
