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Polysome Profiling of mAb Producing CHO Cell Lines Links Translational Control of Cell Proliferation and Recombinant mRNA Loading onto Ribosomes with Global and Recombinant Protein Synthesis

Godfrey, Charlotte, Mead, Emma J, Daramola, Olalekan, Dunn, Sarah, Hatton, Diane, Field, Ray, Pettman, Gary, Smales, Christopher Mark (2017) Polysome Profiling of mAb Producing CHO Cell Lines Links Translational Control of Cell Proliferation and Recombinant mRNA Loading onto Ribosomes with Global and Recombinant Protein Synthesis. Biotechnology Journal, 12 (8). p. 1700177. ISSN 1860-6768. E-ISSN 1860-7314. (doi:10.1002/biot.201700177) (KAR id:61820)

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Abstract

mRNA translation is a key process determining growth, proliferation and duration of a Chinese hamster ovary (CHO) cell culture and influences recombinant protein synthesis rate. During bioprocessing, CHO cells can experience stresses leading to reprogramming of translation and decreased global protein synthesis. Here we apply polysome profiling to determine reprogramming and translational capabilities in host and recombinant monoclonal antibody-producing (mAb) CHO cell lines during batch culture. Recombinant cell lines with the fastest cell specific growth rates were those with the highest global translational efficiency. However, total ribosomal capacity, determined from polysome profiles, did not relate to the fastest growing or highest producing mAb cell line, suggesting it is the ability to utilise available machinery that determines protein synthetic capacity. Cell lines with higher cell specific productivities tended to have elevated recombinant heavy chain transcript copy numbers, localised to the translationally active heavy polysomes. The highest titre cell line was that which sustained recombinant protein synthesis and maintained high recombinant transcript copy numbers in polysomes. Investigation of specific endogenous transcripts revealed a number that maintained or reprogrammed into heavy polysomes, identifying targets for potential cell engineering or those with 5? untranslated regions that might be utilised to enhance recombinant transcript translation.

Item Type: Article
DOI/Identification number: 10.1002/biot.201700177
Uncontrolled keywords: polysome profiling, Chinese hamster ovary (CHO) cells, recombinant monoclonal antibody, cell proliferation, ribosome loading
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Mark Smales
Date Deposited: 23 May 2017 15:26 UTC
Last Modified: 29 May 2019 19:05 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/61820 (The current URI for this page, for reference purposes)
Smales, Christopher Mark: https://orcid.org/0000-0002-2762-4724
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