CD BioGlyco has established a first-class Glycoengineering Platform. We provide clients with efficient antibody remodeling services with G0, G1, G2, or G2S2 glycoforms.

Fc Glycan Remodeling

The conserved asparagine (Asn) at position 297 is glycosylated in the CH2 constant domain of the crystallizable fragment (Fc) region of immunoglobulin G (IgG). N-Glycans are important players in Fc effector function and their composition and structure affect effector function by causing conformational changes in the Fc domain. Therefore, Fc glycan remodeling by glycoengineering is a promising strategy for developing therapeutic antibodies with desired function and efficacy.

• Cell glycoengineering

Cell glycoengineering is the modification of Fc glycans of antibodies by modifying important mediators in the glycan biosynthesis pathway. Currently, there are 3 commonly used methods: 1) Selection of host cell types, environmental factors, and cell culture conditions to produce glycoproteins of desired glycoforms; 2) Modification of host biosynthetic pathways by enzyme inhibitors to obtain target antibodies; 3) Genetically modification of host glycan biosynthetic pathways to obtain glycoproteins.

• Chemoenzymatic glycoengineering

Chemoenzymatic glycoengineering provides a new approach to remodeling heterogeneous N-glycans into homogeneous Fc glycoform. This method deglycosylates the antibody Fc glycans by specific endoglycosidases to leave the innermost layer of N-acetylglucosamine (GlcNAc). The glycosynthase then catalyzes the reaction of the oxazoline of the target glycan with GlcNAc to generate the homologous Fc glycan.

• Site-Specific Antibody-Drug Conjugation (ADC)

ADC is a powerful agent for delivering toxins to target cells. Fc glycans are conserved in all IgGs. Therefore, Fc glycan remodeling is designed for site-specific antibody conjugation to obtain structurally well-defined homogeneous ADC. Glycan heterogeneity can be removed by trimming or extending glycan residues to generate homogeneous ADCs.

Antibody Remodeling Service at CD BioGlyco

CD BioGlyco has developed advanced strategies for Antibody Glycoengineering. We provide clients with efficient antibody remodeling services by chemoenzymatic glycoengineering technology. We efficiently remodel specific human IgG (IgG1, IgG2, and IgG4) glycans at the subunit level using predefined and homogenous G0, G1, G2, or G2S2 glycoforms. Our method trims all Fc glycoforms into core GlcNAc by using specific endoglycosidase enzymes. Glycosynthase catalyzes the transglycosylation reaction between exposed core GlcNAc and oxazoline-reactive G0, G1, G2, or G2S2 glycan.

• Initial Consultation & Antibody Assessment

The process begins with an in-depth consultation to understand your specific therapeutic goals and the characteristics of your antibody. We then perform a comprehensive analysis of the antibody's existing glycoprofile using advanced analytical techniques.

• Enzymatic Glycan Trimming to Core GlcNAc

Our method initiates the remodeling process by precisely trimming all existing Fc glycoforms on the antibody. We use highly specific endoglycosidase enzymes, which cleave the complex glycan structures, leaving behind a uniform core GlcNAc residue exposed on the Fc region.

• Glycan Transglycosylation with Glycosynthase

Following the trimming step, a glycosynthase enzyme is employed to catalyze a highly efficient transglycosylation reaction. This reaction precisely attaches a desired oxazoline-reactive glycan (such as G0, G1, G2, or G2S2) to the exposed core GlcNAc on the antibody Fc region. This chemoenzymatic approach ensures the site-specific and homogeneous incorporation of the target glycoform, resulting in an antibody population with a uniform and predefined glycosylation pattern.

• Purification and Quality Control

After the enzymatic remodeling, the modified antibodies undergo a rigorous purification process to remove enzymes, unreacted substrates, and any byproducts. The purified antibodies are then subjected to extensive quality control, including re-analysis of the glycoprofile by mass spectrometry (MS) and high-performance liquid chromatography (HPLC), to confirm the successful and accurate remodeling of the glycans to the target structure and to ensure high purity.

Workflow

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Applications

• Therapeutic antibody development: Critical for optimizing the efficacy and safety of novel antibody drugs targeting cancer, autoimmune diseases, infectious diseases, and inflammatory conditions.
• Drug discovery and optimization: Facilitates the screening and selection of antibody candidates with superior functional properties early in the discovery pipeline. It allows for rapid optimization of lead candidates for improved in vivo performance.
• Fundamental glycobiology research: Provides a powerful platform for investigating the structure-function relationships of antibody glycans, contributing to a deeper understanding of immunological processes and disease mechanisms.

Advantages

• Our enzymatic approach allows for highly specific and controlled modification of antibody glycans, ensuring the generation of homogeneous glycoforms with defined structures.
• Tailored glycan profiles contribute to a reduction in unwanted immune responses, minimizing the risk of adverse reactions and enhancing the safety profile of the antibody.
• Every remodeled antibody undergoes rigorous characterization using advanced MS and chromatographic techniques, ensuring the accurate and reproducible generation of the desired glycoforms.

Frequently Asked Questions

At CD BioGlyco, our antibody glycan remodeling service is designed to enhance the efficacy, safety, and pharmacokinetic properties of your biotherapeutics, providing a crucial advantage in a competitive landscape. We invite you to contact us to discuss your specific project requirements, explore customized solutions, and learn more about how our services accelerate your research and development efforts.

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