Hi-Affi™ Fucose Knockout Technology Platform
The development of monoclonal antibodies (mAbs) production technology enabled their application in the clinic for the treatment of a wide range of diseases, especially cancer. As the long-term pioneer as well as market leader in the field of therapeutic antibody development, Creative Biolabs has successfully established the fucose knockout technology platform to modulate antibody-dependent cell-mediated cytotoxicity (ADCC).
Background of Our Platform
Along with the clinical success of recombinant humanized therapeutic antibodies against various human malignancies, such as colon, breast, and hematological cancer, the antibody drugs present a major new class in therapeutic agents. However, the improvement of antibody in vivo efficacy continues to be a challenge. There are two independent mechanisms for the in vivo physiological activity mediation, which are target neutralization or apoptosis and biological activities, antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). The importance of ADCC for antibody clinical efficacy has been clear from genetic analyses of leukocyte receptor (FcγR) polymorphisms. It has been proved that compared with fucosylated counterparts, the antibodies lacking core fucose residues from the Fc N-glycans always show stronger ADCC even at lower concentrations. Unfortunately, the existing market therapeutic antibodies are heavily fucosylated as they are produced in mammalian cell lines with intrinsic enzyme activity responsible for the core-fucosylation of the Fc N-glycans of the products. In this case, scientists at Creative Biolabs has established the novel fucose knockout technology platform to improve the antibody ADCC activity.
Fig.1 The impact of fucose and sialic acid on the pharmacokinetic and pharmacodynamic properties of therapeutic IgG.1
Methods for Non-fucosylated Therapeutics
Based on years of research, Creative Biolabs has built several methods for the successful production of highly non-fucosylated therapeutics.
- Non-mammalian N-glycosylation pathway converts to mammalian type.
- Modification of fucosylation pathways in mammalian host cells.
- In vitro fucosylation control system.
Hi-Affi™ Fucose Knockout Technology Platform at Creative Biolabs
Research has reported that the deletion of the FUT8 gene could enhance the FcγRIIIa binding affinity and lead to an increased ADCC activity. FUT8 protein is the only α-1,6 fucosyltransferase that catalyzes the transfer of fucose residues from GDP-fucose to the innermost GlcNAc of the tri-mannosyl core structure via the α-1,6 linkage in the medial Golgi cisternae. Consequently, cell lines that are genetically engineered are incapable of adding fucose to the oligosaccharide chain. The FUT8 knockout has the essential characteristics of host cells for robust manufacturing of fucose-negative therapeutic antibodies with enhanced ADCC, making it an ideal technology to produce ADCC-enhanced antibodies.
Our Platform Advantages
- Enhanced antibody-dependent cellular cytotoxicity (ADCC);
- Eliminating non-human fucose linkages to reduce untargeted immunogenicity;
- Improved efficacy of biopharmaceuticals;
- Streamlined development process to accelerate your therapeutic development projects;
- Customizable fucose knockout approaches to target specific fucosyltransferase genes.
Data Display
| Background | This study focuses on eliminating fucose from target cells, a prevalent system in biopharmaceutical production. The presence of core fucose significantly influences protein function, notably ADCC. Achieving afucosylation, typically through the targeted inactivation of fucosyltransferase genes in the guanine diphosphate-fucose synthesis pathway, represents a widely adopted glycoengineering strategy for developing biologics. |
| Method | This study employs advanced knockout technologies to precisely disrupt the fucosyltransferase gene within the target cell line. |
| Results | The fucosyltransferase gene knockout in target cells leads to a drastic reduction in core-fucosylated glycans across all analyzed sites, including CD166. This modification also broadly impacts the cellular glycosylation landscape, decreasing high-mannose and fucosylated glycans while increasing sialylated forms, and results in a lower overall ratio of core-fucosylated proteins. Additionally, the knockout affected the expression of key glycosylation machinery components (RPN1 and RPN2), indicating a widespread alteration of N-glycan biosynthesis. |
Fig.2 Glycoprotein distribution and identification in fucosyltransferase gene Knock-out (KO) and wild-type (WT) target cells.2
Fig.3 Glycoprotein quantification in fucosyltransferase gene-KO and WT target cells.2
FAQs
Q1: How does the Hi-Affi™ fucose knockout technology platform work?
A1: The platform likely utilizes advanced genetic engineering techniques to specifically knock out or downregulate the genes encoding fucosyltransferase enzymes within the host cell. This prevents fucose from being added to the expressed recombinant protein's N-glycans.
Q2: Which host cell systems can be used with this platform?
A2: Based on our general offerings, this platform likely supports various mammalian and non-mammalian expression systems, including different plant species, yeast strains, and insect cell lines. Please contact our team to acquire the detailed information.
As a pioneer and the undisputed global leader in the field of antibody functional assays, Creative Biolabs offers a series of services and platforms to strengthen your understanding of your candidate antibodies and to facilitate their transformation from preclinical to the clinic. If you are interested in our technologies and services, please do not hesitate to contact us for more detailed information.
References
- Luo, Cheng, et al. "Glycoengineering of pertuzumab and its impact on the pharmacokinetic/pharmacodynamic properties." Scientific Reports 7.1 (2017): 46347. Distributed under Open Access License CC BY 4.0, without modification.
- Yang, Ganglong, et al. "Glycoproteomic characterization of FUT8 knock-out CHO cells reveals roles of FUT8 in the glycosylation." Frontiers in Chemistry 9 (2021): 755238. Distributed under Open Access License CC BY 4.0, without modification.
For Research Use Only.