Genetic engineering of α2,6-sialyltransferase in recombinant CHO cells and its effects on the sialylation of recombinant interferon-γ.

Cytotechnology
L MonacoN Jenkins

Abstract

The CHO cell line has achieved considerable commercial importance as a vehicle for the production of human therapeutic proteins, but is known to lack a functional copy of the gene coding for α2,6-sialyltransferase (EC 2.4.99.1). The cDNA for rat α2,6-ST was expressed in a recombinant CHO cell line making interferon-γ, using a novel in vitro amplification vector. The enzyme was expressed efficiently, and resulted in up to 60% of the total sialic acids on interferon-γ being linked in the α2,6-conformation. This sialic acid linkage distribution was more akin to that seen in natural human glycoproteins. In the most successful cell clones, expression of α2,6-sialyltransferase improved the overall level of sialylation by up to 56%, and had no adverse effects on cell growth, IFN-γ productivity or other aspects of IFN-γ glycosylation. These experiments demonstrate how the glycosylation machinery of rodent cells can be genetically manipulated to replicate human tissues.

References

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Citations

Apr 13, 2002·Current Opinion in Biotechnology·Dana C Andersen, Lynne Krummen
Aug 14, 2001·Biochemical and Biophysical Research Communications·R JassalJ Lund
Apr 16, 2010·Biotechnology and Applied Biochemistry·Yiping LimMiranda G S Yap
Nov 20, 2018·Frontiers in Immunology·Matthew J BuettnerKevin J Yarema
Feb 3, 2021·Protein Expression and Purification·Zhongcheng ZouPeter D Sun

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