Production of α2,6-sialylated IgG1 in CHO cells

@article{Raymond2015ProductionO,
  title={Production of $\alpha$2,6-sialylated IgG1 in CHO cells},
  author={C{\'e}line Raymond and Anna C Robotham and Maureen A. Spearman and Michael Butler and John F Kelly and Yves Durocher},
  journal={mAbs},
  year={2015},
  volume={7},
  pages={571 - 583}
}
The presence of α2,6-sialic acids on the Fc N-glycan provides anti-inflammatory properties to the IgGs through a mechanism that remains unclear. Fc-sialylated IgGs are rare in humans as well as in industrial host cell lines such as Chinese hamster ovary (CHO) cells. Facilitated access to well-characterized α2,6-sialylated IgGs would help elucidate the mechanism of this intriguing IgG's effector function. This study presents a method for the efficient Fc glycan α2,6-sialylation of a wild-type… 

Production of IgGs with a human-like sialylation in CHO cells

In this study, it is shown that the a2,6-sialylation of IgG1’s Fc domain can be efficiently achieved by the transient coexpression of the human b1,4-galactosyltransferase 1 (GT) and a3,3-sIALyltransferases 1 (ST6) in CHO cells, whereas the expression of one or the other glycosyl transferase alone does not significantly improve sialylations.

Impact of Fc N-glycan sialylation on IgG structure

Investigating the impact of different types of sialylation to the conformational stability of IgG through hydrogen/deuterium exchange and limited proteolysis experiments found only alpha 2,3-linked sialic acid on the 6-arm destabilizes the CH2 domain, presumably because of the steric effect that decreases the glycan-CH2 domain interaction.

Enhanced sialylation of a human chimeric IgG1 variant produced in human and rodent cell lines.

Impact of IgG1 N-glycosylation on their interaction with Fc gamma receptors

Integrated Genome and Protein Editing Swaps α-2,6 Sialylation for α-2,3 Sialic Acid on Recombinant Antibodies from CHO.

Engineered antibodies from these modified Chinese hamster ovary cell lines will provide biotechnologists with IgGs containing N-glycans with different structural variations for examining the role of glycosylation on protein performance.

Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans

General methods to produce recombinant proteins of any desired glycoform in eukaryotic cells are described and the biological effect of these glycosylation traits for IgG and other glycoproteins are systematically explored.

Metabolic engineering of CHO cells to prepare glycoproteins.

The interplay of multiple glyco-gene knockouts and knockins and the combination of multiple approaches, including genetic manipulation, protein engineering and chemical supplementation, are detailed in order to achieve specific glycan profiles on recombinant glycoproteins for superior biological function and effectiveness.

Overexpression of α (1,6) fucosyltransferase in the development of castration-resistant prostate cancer cells

This is the first study reporting the functional role of fucosylated enzyme in the development of castration-resistant prostate cancer and demonstrating that overexpression of FUT8 might be responsible for the decreased PSA expression in prostate cancer specimens.
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