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Overexpression

Gene overexpression could be applied to test specific hypotheses and to identify the hypothesized targets. Based on the advanced gene overexpression technology, Creative Biolabs is providing gene overexpression services for our clients from all over the world, especially the clients who are currently having difficulty working on cell line glycoengineering.

Background of Gene Overexpression

Gene overexpression technique provides a powerful alternative tool to infer gene functions that remain to be discovered by gene knockout analysis or not yet possible creating targeted deletions. Novel biotherapeutic glycoproteins, like recombinant monoclonal antibodies (mAbs) are widely used for the treatment of numerous diseases. In glycoengineering, desired properties like the addition of sialic acid residues to a growing N-glycan chain, increasing triantennary structures, introducing non-fucosylated hybrid-type glycans, increased antibody effector function could be achieved by gene overexpression.

Application in Glycoengineering

Among different glycoprotein production systems, gene overexpression is widely used in the construction of engineered Chinese hamster ovary (CHO) cells. Overexpression of GalT produces an increase in the sialic acid content in CHO cells. For recombinant glycoprotein production, overexpression of CMP-sialic acid transporter (CMP-SAT) in CHO cells expressing recombinant human interferon gamma (IFN-γ) increased IFN-γ sialylation by 4-16%. Overexpression of N-sulfotransferase 2 (NDST2) and heparan sulfate 3-O-sulfotransferase 1 (3OST1) in CHO cells increase the amount of glycosaminoglycan (GAGs) secreted into the culture medium by 10 fold. Additionally, CHO cells overexpressing 30Kc19 gene not only increased rhEPO sialylation by 87.1% but also increased its yield by 102.6%. Bisecting GlcNAc glycoforms and increased antibody effector function could be achieved by the overexpression of β-1,4-GlcNAc transferase III (GnT III) and this technology has been used to produce the FDA approved anti-CD20 obinutuzumab. Triantennary structures were significantly increased by overexpression of GnT-IV or GnT-V individually in CHO cells. Co-overexpression of GnT III and α-mannosidase II (Man II) leads to further lower fucose content by introducing non-fucosylated hybrid-type glycans. Lastly, the altered glycan ratios can be overcome by overexpressing ST6Gal 1 in CHO cells.

Composition of a complex oligosaccharide attached to IgG Fc Fig.1 Composition of a complex oligosaccharide attached to IgG Fc (Popp, 2018).

Numerous engineered E. coli strains have also been engineered. For instance, there is a non-pathogenic E. coli K12 strain that overexpresses recombinant sialyltransferases and sialic acid synthesis genes, which can convert an exogenous lactoside into polysialyl lactosides.

Features of our Overexpression Services

  • Effective overexpression vector for cell line glycoengineering
  • Complete project report
  • Perfect after-sale service system
  • Therapeutic glycosylated antibodies with a higher ADCC function
  • Cost-effective and time-saving

With our rich experience in offering gene overexpression services, scientists of Creative Biolabs are pleased to tailor the best-fit research program for your purpose. If you are considering using the gene overexpression for cell line glycoengineering, please do not hesitate to contact us for more details.

Reference

  1. Popp, O.; et al. Development of a pre-glycoengineered CHO-K1 host cell line for the expression of antibodies with enhanced Fc mediated effector function. Mabs. 2018, 10(2), 290-303.
For Research Use Only.

Related Services:

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