Cell Glycoengineering by Manipulating Branching
Biosynthesis of N-glycan Branching in Mammals
In mammalian cells, the process of N-glycan branching occurs within the medial Golgi and is controlled by specific enzymes known as GnT-I, GnT-II, GnT-IV, and GnT-V. GnT-I and GnT-II are responsible for generating bi-antennary N-glycan structures, and the formation of tri- and tetra-antennary N-glycans is regulated by GnT-IV and GnT-V. These branched structures can further be extended in the trans-Golgi by additional enzymes capable of adding sugar residues like Gal, Fuc, and sialic acids.
Fig.1 Biosynthesis pathway of branching N-glycans.1
Cell Glycoengineering by Manipulating Branching at Creative Biolabs
Being at the forefront of glycoengineering services, Creative Biolabs has established robust technology platforms and a range of strategies to control glycoforms like Heterogeneity, Sialylation, and Fucosylation in different expression systems. To modulate N-glycan branching, we employ genetic techniques, such as Overexpression of enzymes responsible for the biosynthesis of branching N-glycans with desired structures.
Advanced techniques
Our technology platforms encompass advanced techniques and tools for gene overexpression. We are experienced in developing transient overexpression systems in various cell lines using tailored expression vectors. For stable overexpression, we utilize lentiviral transduction, which allows for the stable integration of highly customizable vectors and is particularly effective for difficult-to-transfect cells. Additionally, we provide precision genome editing techniques, enabling site-directed and stable knockin of target genes.
Comprehensive strategies
Overexpression of enzymes, such as GnT-IV and GnT-V, enhances N-glycan branching. By independently or simultaneously overexpressing these enzymes, N-glycans featuring tri-antennary or tetra-antennary structures can be effectively produced in engineered cell lines.
Applications of Manipulating Branching
The branching of N-glycans plays a pivotal role in determining the biological activity and circulatory lifespan of recombinant therapeutic glycoproteins. Increasing the N-glycan branching in therapeutic glycoproteins can result in the production of higher levels of sialic acid content, leading to an extended half-life in circulation. By modulating the branching patterns of N-glycans, researchers can finely tune the properties of therapeutic glycoproteins to optimize their efficacy and pharmacokinetic profiles, thereby enhancing their therapeutic potential.
Advantages of Our Services
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Optional genetic strategies for tailored N-glycan structures
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Advanced and optional techniques for the regulation of gene expression
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Effective expression vector for high-level expression of target gene
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Customized cell glycoengineering for glycan branching
Creative Biolabs stands out as a pioneering leader in genetic glycoengineering services. Our commitment to advancing glycoengineering enables us to offer tailored solutions for the modulation of glycan branching. Please feel free to contact us to explore how we can elevate your glycoengineering projects.
Reference
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Tejwani, Vijay, et al. "Glycoengineering in CHO cells: advances in systems biology." Biotechnology Journal 13.3 (2018): 1700234.
For Research Use Only.
Related Services:
- Knockdown
- Overexpression
- Knockout and Knockin by Precision Genome Editing
- Manipulating Heterogeneity
- Manipulating Sialylation
- Manipulating Fucosylation
- Manipulating Glycosaminoglycan
