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Cell Pool Selection of CHO Host and Recombinant Cell Pools by Inhibition of the Proteasome Results in Enhanced Product Yields and Cell Specific Productivity

Knight, Tanya J, Turner, Sarah, Jaques, Colin M, Smales, Christopher Mark (2021) Cell Pool Selection of CHO Host and Recombinant Cell Pools by Inhibition of the Proteasome Results in Enhanced Product Yields and Cell Specific Productivity. Journal of Biotechnology, 337 . pp. 35-45. ISSN 0168-1656. (doi:10.1016/j.jbiotec.2021.06.019) (KAR id:89634)

Abstract

Chinese hamster ovary (CHO) cells are the leading mammalian cell expression platform for biotherapeutic recombinant molecules yet some proteins remain difficult to express (DTE) in this, and other, systems. In recombinant cell lines expressing DTE proteins, cellular processes to restore proteostasis can be triggered when the folding and modification capabilities are exceeded, including the unfolded protein response and ER associated degradation (ERAD) and proteasomal degradation. We therefore investigated whether the proteasome activity of CHO cells was linked to their ability to produce recombinant proteins. We found cell lines with diverse monoclonal antibody (mAb) productivity show different susceptibilities to inhibitors of proteasome activity. Subsequently, we applied selective pressure using proteasome inhibitors on mAb producing cells to determine the impact on cell growth and recombinant protein production, and to apply proteasome selective pressure above that of a metabolic selection marker during recombinant cell pool construction. The presence of proteasome inhibitors during cell pool construction expressing two different model molecules, including a DTE Fc fusion protein, resulted in the generation of cell pools with enhanced productivity. The increased productivities, and ability to select for higher producing cells, has potential to improve clonal selection during upstream processes of DTE proteins.

Item Type: Article
DOI/Identification number: 10.1016/j.jbiotec.2021.06.019
Uncontrolled keywords: Proteasome inhibitors, Chinese hamster ovary cells, ERAD, Recombinant protein production, Difficult to express proteins
Subjects: Q Science
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Mark Smales
Date Deposited: 05 Aug 2021 10:27 UTC
Last Modified: 14 Nov 2022 23:13 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/89634 (The current URI for this page, for reference purposes)

University of Kent Author Information

Smales, Christopher Mark.

Creator's ORCID: https://orcid.org/0000-0002-2762-4724
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