Cell Glycoengineering by Manipulating Fucosylation

Core Fucosylation Pathway in Mammals

Core fucosylation is a widespread and significant characteristic of N-glycosylation. The core fucose is linked via an α-1,6 linkage to the innermost GlcNAc residue within the N-oligosaccharide. In mammals, FUT8 stands as the sole α-1,6 fucosyltransferase responsible for transferring fucose residues. GDP-fucose serves as the donor substrate for fucose transfer during glycosylation. The synthesis of GDP-fucose occurs in the cytoplasm through two distinct pathways: the de novo pathway and the salvage pathway. In the de novo pathway, GDP-mannose is converted by GDP-mannose 4,6-dehydratase (GMD). The salvage pathway involves the direct utilization of fucose. Subsequently, the cytoplasmic GDP-fucose is transported into the Golgi apparatus by the GDP-fucose transporter (GFT).

Fig.1 Core fucosylation pathway in mammalian cells.¹

Cell Glycoengineering by Manipulating Fucosylation at Creative Biolabs

Equipped with advanced platforms in Glycoengineering Services, Creative Biolabs provides multiple genetic interventions to manipulate fucosylation in diverse expression systems. We use various methods such as antisense cDNA, small interfering RNAs (siRNAs), or transgenic overexpression to inhibit or eliminate core fucosylation. To enhance the efficiency of gene disruption, technologies like TALENs and the CRISPR/Cas9 system are also modified in our laboratory to induce targeted genetic Knockout and Knockin. These modulations are mainly targeted at the following pathways.

In addition to genetic modulations in mammalian cells, we also provide services to modify fucosylation in Plant Cells and Insect Cells by conversion of non-mammalian N-glycosylation pathway into the mammalian type.

Glycoengineering for the Removal of Core Fucose: Applications and Benefits

The absence of core fucose has a significant impact on the efficacy and safety of therapeutic proteins. N-glycans containing fucose located at Asn297 in IgG antibody detrimentally influence the Fc-FcɣRIII interaction. Elimination of core fucose from human IgG enhances the binding affinity of the Fc domain to FcɣRIII and results in a remarkable increase of up to 100-fold in antibody-dependent cellular cytotoxicity (ADCC).

Advantages of Our Services

• Optional genetic strategies for manipulating fucosylation
• Cutting-edge technologies for precise genome editing
• Effective manipulation of fucosylation across different expression systems
• Professional team with extensive experience in glycoengineering

Published Data

Technology: Antibody glycoengineering

Journal: mAbs

IF: 6.44

Published: 2009

Results: Non-fucosylated antibodies have revolutionized antibody therapies by significantly enhancing ADCC. One of the most feasible and reliable strategies for manufacturing fully non-fucosylated therapeutic antibodies is by using FUT8 knockout CHO/DG44 cells as the production system.

Fig.2 Non-fucosylated antibodies induce high ADCC.²

Creative Biolabs, as a leading company in the field of glycoengineering, has acquired extensive experience in fucosylation-targeted glycoengineering in various expression systems. To obtain more details, please don't hesitate to contact us or directly submit an inquiry.

References

1. Sun, Yuhan, et al. "Core fucosylation regulates the function of pre-BCR, BCR and IgG in humoral immunity." Frontiers in Immunology 13 (2022): 844427.
2. Yamane-Ohnuki, Naoko, and Mitsuo Satoh. "Production of therapeutic antibodies with controlled fucosylation." MAbs. Vol. 1. No. 3. Taylor & Francis, 2009.

Related Services:

1. Knockdown
2. Overexpression
3. Knockout and Knockin by Precision Genome Editing
4. Manipulating Heterogeneity
5. Manipulating Sialylation
6. Manipulating Branching
7. Manipulating Glycosaminoglycan