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Effect of process parameters and product-host-interaction on hVEGFA-production by recombinant Chinese hamster ovary cells

Titelangaben

Freimark, Denise ; Jérôme, Valérie ; Freitag, Ruth:
Effect of process parameters and product-host-interaction on hVEGFA-production by recombinant Chinese hamster ovary cells.
In: Biotechnology Progress. Bd. 28 (Mai 2012) Heft 3 . - S. 762-772.
ISSN 1520-6033
DOI: https://doi.org/10.1002/btpr.1524

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Abstract

A potential producer clone was identified among recombinant, human vascular endothelial growth factor A (hVEGFA)-producing Chinese Hamster Ovary (CHO) K1 cells, using a recently established screening method. In batch spinner cultivations, the cells showed a maximum growth rate of 0.045 h(-1), a final total cell density of 5.3×10(6) mL(-1) (living cell density: 3.4×10(6) mL(-1)), and a final hVEGFA concentration of 207 μg L(-1). Living cell density and productivity in the spinner cultivations could be increased by glutamine feeding. Transfer of the process to the bioreactor (batch mode, control of pH, T, and O2) resulted in a reduction of the growth rate by roughly 50%, while overall living cell density and productivity increased, largely due to an extension of the production phase. When the bioreactor was run in the fed-batch mode, growth rates were further reduced, while productivity and living cell densities reached a maximum (hVEGFA: 358 μg L(-1), cells: 5.2×10(6) mL(-1)). In addition, the death rate of the hVEGFA-producing cells was considerably reduced compared with the parent cell line, most likely due to product-host-interaction. This hypothesis was corroborated when a second recombinant CHO cell line (antibody producer) was transfected with the hVEGFA gene and afterward consistently showed higher viable cell densities together with a significantly improved antibody titer.

Weitere Angaben

Publikationsform: Artikel in einer Zeitschrift
Begutachteter Beitrag: Ja
Keywords: bioprocess engineering; fed-batch; growth factor; mammalian cell culture; recombinant protein production
Institutionen der Universität: Fakultäten
Fakultäten > Fakultät für Ingenieurwissenschaften
Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik
Fakultäten > Fakultät für Ingenieurwissenschaften > Lehrstuhl Bioprozesstechnik > Lehrstuhl Bioprozesstechnik - Univ.-Prof. Dr. Ruth Freitag
Titel an der UBT entstanden: Ja
Themengebiete aus DDC: 600 Technik, Medizin, angewandte Wissenschaften
600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin und Gesundheit
600 Technik, Medizin, angewandte Wissenschaften > 620 Ingenieurwissenschaften
Eingestellt am: 23 Feb 2016 15:06
Letzte Änderung: 29 Feb 2016 08:26
URI: https://eref.uni-bayreuth.de/id/eprint/31023