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Gewählte Publikation:

Taschwer, M; Hackl, M; Hernández Bort, JA; Leitner, C; Kumar, N; Puc, U; Grass, J; Papst, M; Kunert, R; Altmann, F; Borth, N.
(2012): Growth, productivity and protein glycosylation in a CHO EpoFc producer cell line adapted to glutamine-free growth.
J Biotechnol. 2012; 157(2):295-303 FullText FullText_BOKU

A primary objective of cell line development and process optimisation in animal cell culture is the improvement of culture performance as indicated by desirable properties such as high cell concentration, viability, productivity and product quality. The inefficient energy metabolism of mammalian cells in culture is still a major limiting factor for improvements in process performance. It results in high uptake rates of glucose and glutamine and the concomitant accumulation of waste products which in turn limits final cell concentrations and growth. To avoid these negative side effects, a CHO host cell line was established recently which is able to grow in completely glutamine free medium (Hernandez Bort et al.. 2010). To determine the influence of this adaptation on productivity and product quality, the same procedure was repeated with a recombinant CHO cell line producing an erythropoietin-Fc fusion protein (CHO-EpoFc) for this publication. After adaptation to higher cell densities and glutamine free medium, culture performance was monitored in batch bioprocesses and revealed comparable growth properties and EpoFc product formation in both cell lines. The level of reactive oxygen species was elevated in the adapted cells, reflecting a higher level of oxidative stress, however, at the same time the level of the oxido-protective glutathione was also higher, so that cells seem adequately protected against cellular damage. Analysis of nucleotides and nucleotide sugars revealed elevated UDP-sugars in cells grown in the absence of glutamine. Furthermore, the antennarity of N-glycans was moderately higher on the Epo part of the protein produced by the adapted cell line compared to the parental cell line. Except for this, the glycosylation, with respect to site occupancy, degree of sialylation and glycoform structure, was highly comparable, both for the Epo and the Fc part of the protein. (C) 2011 Elsevier B.V. All rights reserved.
Autor*innen der BOKU Wien:
Altmann Friedrich
Borth Nicole
Grass Josephine
Hackl Matthias
Hernandez-Bort Juan-Antonio
Kunert Renate
Leitner Christian

Find related publications in this database (Keywords)
High density
Glutamine free
Cell sorting
Oxidative stress

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