Bioreactor cultivation of CHO DP-12 cells under sodium butyrate treatment – comparative transcriptome analysis with CHO cDNA microarrays

Klausing S, Krämer O, Noll T. Bioreactor cultivation of CHO DP-12 cells under sodium butyrate treatment – comparative transcriptome analysis with CHO cDNA microarrays. BMC Proceedings. 2011;5(Suppl 8)

On the AAVenue to success: Advances in technologies for AAV production

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Optimierung von CHO Produktionszelllinien: RNAi-vermittelter Gen-knockdown und Untersuchungen zur Klonstabilität

Klausing S. Optimierung von CHO Produktionszelllinien: RNAi-vermittelter Gen-knockdown und Untersuchungen zur Klonstabilität. Bielefelder Schriften zur Zellkulturtechnik. Vol 10. Berlin: Logos; 2013

Methods in mammalian cell line engineering: from random mutagenesis to sequence specific approaches

Krämer O, Klausing S, Noll T. Methods in mammalian cell line engineering: from random mutagenesis to sequence specific approaches. Applied Microbiology and Biotechnology. 2010;88(2):425-436.Due to the increasing demand for recombinant proteins, the interest in mammalian cell culture, especially of Chinese hamster ovary cells, grows rapidly. This is accompanied by the desire to improve cell lines in order to achieve higher titers and a better product quality. Until recently, most cell line development procedures were based on random integration and gene amplification, but several methods for targeted genetic modification of cells have been developed. Some of those are homologous recombination, RNA interference and zinc-finger nucleases. Especially the latter two have evolved considerably and will soon become a standard for cell line engineering in research and industrial application. This review presents an overview of established as well as new and promising techniques for targeted genetic modification of mammalian cells

Perfusion process combining low temperature and valeric acid for enhanced recombinant factor VIII production.

Coronel J, Heinrich C, Klausing S, Noll T, Figueredo-Cardero A, Castilho LR. Perfusion process combining low temperature and valeric acid for enhanced recombinant factor VIII production. Biotechnology progress. 2019;36(1):e2915.Perfusion operation mode remains the preferred platform for production of labile biopharmaceuticals (e.g., blood factors) and is also being increasingly adopted for production of stable products (e.g., monoclonal antibodies). Regardless of the product, process development typically aims at maximizing production capacity. In this work, we investigated the impact of perfusion cultivation conditions on process productivity for production of human factor VIII (FVIII). Recombinant CHO cells were cultivated in bioreactors coupled to inclined settlers and the effects of reducing the temperature to 31°C with or without valeric acid (VA) supplementation were evaluated. Increases in cell specific productivity (qp ) up to 2.4-fold (FVIII concentration) and up to 3.0-fold (FVIII biological activity) were obtained at 31°C with VA compared to the control at 37°C. Biological activity is the most important quality attribute for FVIII and was positively affected by mild hypothermia in combination with the chemical inducer. The low temperature conditions resulted in enhanced product transcript levels, suggesting that the higher qp is related to the increased mRNA levels. Furthermore, a high-producer subclone was evaluated under the perfusion conditions optimized for the parental clone (31°C with VA), yielding increases in qp of 6-fold and 15-fold compared to the parental clone cultivated under the same condition and at 37°C, respectively. The proposed perfusion strategy enables increased product formation without increasing production costs, being potentially applicable to perfusion production of other CHO-derived biopharmaceuticals. To the best of our knowledge, this is the first report showing the benefits of perfusion combining mild hypothermia with VA supplementation. This article is protected by copyright. All rights reserved. © 2019 American Institute of Chemical Engineers

Valeric acid supplementation combined to mild hypothermia increases productivity in CHO cell cultivations

Coronel J, Klausing S, Heinrich C, Noll T, Figueredo-Cardero A, Castilho LR. Valeric acid supplementation combined to mild hypothermia increases productivity in CHO cell cultivations. Biochemical Engineering Journal. 2016;114:101-109