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

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.

journal_name

Cytotechnology

journal_title

Cytotechnology

authors

Monaco L,Marc A,Eon-Duval A,Acerbis G,Distefano G,Lamotte D,Engasser JM,Soria M,Jenkins N

doi

10.1007/BF00353939

subject

Has Abstract

pub_date

1996-01-01 00:00:00

pages

197-203

issue

1-3

eissn

0920-9069

issn

1573-0778

journal_volume

22

pub_type

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