CD BioGlyco has developed an advanced Antibody Glycoengineering platform to efficiently remodel antibodies with G0 glycoforms.
The development of recombinant protein therapeutics is a major revolution in modern medicine. Over the past two decades, the number of therapy-based monoclonal antibodies (mAbs) has grown rapidly, bringing new therapeutic hope to cancer patients. Therapeutic mAbs are typically immunoglobulin G (IgG) containing four polypeptide chains. Their conserved asparagine (Asp) is glycosylated at position 297 in the CH2 constant domain of the crystallizable fragment (Fc) region. Fc glycosylation has a great influence on the structure, immunogenicity, pharmacokinetics, and pharmacodynamics of mAb. It has been found that the immunological properties of antibodies can be improved by modulating the fine structure of glycans, especially those linked to the Fc domain. For example, reducing Fc core fucosylation can significantly enhance the antibody-dependent cellular cytotoxicity (ADCC) of therapeutics, thereby improving anticancer activity.
Since different glycans have a significant impact on the function of antibodies. Therefore, researchers have modified the glycan structure in the Fc domain by glycoengineering strategies to improve the therapeutic effect. There are currently two commonly used glycoengineering strategies, namely host cell glycoengineering and in vitro chemoenzymatic glycosylation remodeling. Host cell glycoengineering is the modification of glycan structure by manipulating important mediators in the glycan biosynthetic pathway of different host expression systems. This method has been used to produce optimal glycoforms for some therapeutic antibodies. In vitro chemoenzymatic glycosylation remodeling provides a novel approach to remodeling Fc glycans from a heterogeneous N-glycosylation pattern to a homogeneous pattern. First, deglycosylating IgG antibodies is performed by specific enzymes and leaving the innermost layer of N-Acetylglucosamine (GlcNAc). Glycoengineered mAbs with homologous N-glycans are then prepared by transglycosylation.
Fig.1 In vitro chemoenzymatic glycosylation remodeling using endo-β-N-acetylglucosaminidase and glycosynthase. (Boune, et al., 2020)
Since natural and recombinant antibodies are often produced as a heterogeneous mixture of glycoforms, it is extremely difficult to isolate or enrich for pure glycans. Therefore, it is highly necessary to develop glycoengineering strategies to generate glycan-defined and site-selectively modified antibodies, improving their function and therapeutic efficacy. At CD BioGlyco, we have developed an advanced antibody glycoengineering platform. We provide clients with efficient and site-specific glycan remodeling services for human IgG, including human IgG1, IgG2, and IgG4. We generate human IgG carrying the G0 glycoform by an in vitro chemoenzymatic glycosylation remodeling technique. Both fucosylated G0 and afucosylated G0 glycoform are available. Our workflow is as follows:
1) Trim Fc glycans to the innermost GlcNAc by specific endoglycosidases.
2) Glycosynthase catalyzes the reaction of pre-assembled G0 glycan oxazoline with core GlcNAc to generate antibodies with homologous G0 glycoforms.
Fig.2 Process of remodeling an antibody with G0 Glycoform. (CD BioGlyco)
CD BioGlyco has extensive experience in Remodeling Antibody. We are pleased to share our knowledge and methods. If you are interested in remodeling antibody with G0 glycoforms, please feel free to contact us.
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