Selection of CHO host and recombinant cell pools by inhibition of the proteasome results in enhanced product yields and cell specific productivity.

Autor: Knight TJ; Industrial Biotechnology Centre, School of Biosciences, University of Kent, Canterbury, Kent, CT2 7NJ, UK., Turner S; Lonza Biologics, 228 Bath Road, Slough, SL1 4DX, UK., Jaques CM; Lonza Biologics, 228 Bath Road, Slough, SL1 4DX, UK. Electronic address: colin.jaques@lonza.com., Smales CM; Industrial Biotechnology Centre, School of Biosciences, University of Kent, Canterbury, Kent, CT2 7NJ, UK. Electronic address: c.m.smales@kent.ac.uk.
Jazyk: angličtina
Zdroj: Journal of biotechnology [J Biotechnol] 2021 Aug 20; Vol. 337, pp. 35-45. Date of Electronic Publication: 2021 Jun 24.
DOI: 10.1016/j.jbiotec.2021.06.019
Abstrakt: 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.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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