N-Linked Glycoengineering Service in Insect Cell

N-Glycosylation in Insect and Mammalian Cells

Insect and mammalian cells share a common intermediate structure in N-glycosylation, but the following processes are generally different. In mammalian cells, after the addition of a GlcNAc residue to both mannose branches, various glycosyltransferases elongate the branches to form complex N-glycans. However, insect cells do not elongate both branches. Instead, they express a specific enzyme called fused lobes (FDL), which is absent in mammalian cells. FDL removes the terminal GlcNAc residue from hybrid-type trimannosyl core structures, converting them into smaller and structurally simpler paucimannose N-glycans. These paucimannose N-glycans are characterized by terminal mannose residues and lack terminal GlcNAc, galactose, or sialic acid residues, as well as higher-order branching. Furthermore, insect cells can synthesize specific glycan structures such as core α1,3-fucose catalyzed by the α1,3 fucosyltransferase. These differences emphasize the importance of glycoengineering efforts to humanize glycosylation pathways in insect cells for recombinant glycoprotein production.

N-Linked Glycoengineering Services in Insect Cell at Creative Biolabs

Creative Biolabs has harnessed advanced technologies for manipulating gene and protein expression to establish glycoengineering services in insect cells, with the ultimate goal of producing customized glycoproteins with humanized N-glycan patterns. We offer a range of glycoengineering strategies and comprehensive services to overcome the functional limitations associated with N-glycan processing in insect systems and to remove insect-specific glycan structures. These services encompass a variety of approaches, including but not limited to:

• Introduction of humanized N-glycosylation pathway
  Introduction of enzymes for N-glycan elongation, mainly involving the Overexpression of human GlcNAc-TI/II and GalT-I.
  Introduction of the sialylation pathway by co-expressing sialic acid synthase, CMP-sialic acid synthase and transporter, and ST6Gal1.
• Elimination of undesirable insect-specific glycans
  Suppression or elimination of FDL activity, through siRNA-mediated Silencing or efficient Knockout with gene editing tools.
  Elimination of α1,3-core fucose by knockout of α1,3 fucosyltransferase or expression of GDP-6-deoxy-D-lyxo-4-hexulose reductase (RMD) inhibitor.

Most of these approaches can be applied to either the insect host cells or the baculoviral vectors to enhance their endogenous N-glycan processing capabilities. The optimal strategies and tools will be employed based on a case-by-case assessment. Our commitment to advanced technologies and extensive expertise in glycoengineering ensures that we can meet your specific needs.

Applications of Glycoengineered Insect Expression Systems

• Production of glycoprotein therapeutics with extended half-lives in human sera
• Production of glycoprotein therapeutics with higher safety
• Production of glycoprotein therapeutics with improved therapeutic efficacy

Advantages of Our Services

• Optional glycoengineering strategies encompassing both baculovirus and insect host.
• Cutting-edge techniques for gene regulation and precise genome editing
• High-level gene expression and high-yield glycoprotein production
• Customized glycoengineering for tailored glycan patterns
• Professional team with extensive experience in glycoengineering

As glycoengineering efforts continue to advance, insect cells as a production platform hold significant potential for further growth in the field of glycoprotein production. Creative Biolabs is a leading provider of comprehensive glycoengineering services in insect cells. Please feel free to contact us for more details.