miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering

Stiefel F, Fischer S, Sczyrba A, Otte K, Hesse F (2016)
Journal of Biotechnology 225: 31-43.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
Fed-batch cultivation of recombinant Chinese hamster ovary (CHO) cell lines is one of the most widely used production modes for commercial manufacturing of recombinant protein therapeutics. Furthermore, fed-batch cultivations are often conducted as biphasic processes where the culture temperature is decreased to maximize volumetric product yields. However, it remains to be elucidated which intracellular regulatory elements actually control the observed pro-productive phenotypes. Recently, several studies have revealed microRNAs (miRNAs) to be important molecular switches of cell phenotypes. In this study, we analyzed miRNA profiles of two different recombinant CHO cell lines (high and low producer), and compared them to a non-producing CHO DG44 host cell line during fed-batch cultivation at 37°C versus a temperature shift to 30°C. Taking advantage of next-generation sequencing combined with cluster, correlation and differential expression analyses, we could identify 89 different miRNAs, which were differentially expressed in the different cell lines and cultivation phases. Functional validation experiments using 19 validated target miRNAs confirmed that these miRNAs indeed induced changes in process relevant phenotypes. Furthermore, computational miRNA target prediction combined with functional clustering identified putative target genes and cellular pathways, which might be regulated by these miRNAs. This study systematically identified novel target miRNAs during different phases and conditions of a biphasic fed-batch production process and functionally evaluated their potential for host cell engineering. Copyright 2016. Published by Elsevier B.V.
Erscheinungsjahr
Zeitschriftentitel
Journal of Biotechnology
Band
225
Seite
31-43
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Stiefel F, Fischer S, Sczyrba A, Otte K, Hesse F. miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering. Journal of Biotechnology. 2016;225:31-43.
Stiefel, F., Fischer, S., Sczyrba, A., Otte, K., & Hesse, F. (2016). miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering. Journal of Biotechnology, 225, 31-43. doi:10.1016/j.jbiotec.2016.03.028
Stiefel, F., Fischer, S., Sczyrba, A., Otte, K., and Hesse, F. (2016). miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering. Journal of Biotechnology 225, 31-43.
Stiefel, F., et al., 2016. miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering. Journal of Biotechnology, 225, p 31-43.
F. Stiefel, et al., “miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering”, Journal of Biotechnology, vol. 225, 2016, pp. 31-43.
Stiefel, F., Fischer, S., Sczyrba, A., Otte, K., Hesse, F.: miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering. Journal of Biotechnology. 225, 31-43 (2016).
Stiefel, Fabian, Fischer, Simon, Sczyrba, Alexander, Otte, Kerstin, and Hesse, Friedemann. “miRNA profiling of high, low and non-producing CHO cells during biphasic fed-batch cultivation reveals process relevant targets for host cell engineering”. Journal of Biotechnology 225 (2016): 31-43.

4 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

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Malekpour Afshar R, Mollaei HR, Shokrizadeh M, Iranpour M., Asian Pac J Cancer Prev 18(3), 2017
PMID: 28440610
miRNA engineering of CHO cells facilitates production of difficult-to-express proteins and increases success in cell line development.
Fischer S, Marquart KF, Pieper LA, Fieder J, Gamer M, Gorr I, Schulz P, Bradl H., Biotechnol Bioeng 114(7), 2017
PMID: 28262952
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PMID: 28371547

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