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

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.

Location
Deutsche Nationalbibliothek Frankfurt am Main
Extent
Online-Ressource
Language
Englisch

Bibliographic citation
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 ; volume:12 ; number:8 ; year:2017 ; extent:12
Biotechnology journal ; 12, Heft 8 (2017) (gesamt 12)

Creator
Godfrey, Charlotte L.
Mead, Emma J.
Daramola, Olalekan
Dunn, Sarah
Hatton, Diane
Field, Ray
Pettman, Gary
Smales, C. Mark

DOI
10.1002/biot.201700177
URN
urn:nbn:de:101:1-2022092806243653174690
Rights
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
Last update
15.08.2025, 7:26 AM CEST

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Associated

  • Godfrey, Charlotte L.
  • Mead, Emma J.
  • Daramola, Olalekan
  • Dunn, Sarah
  • Hatton, Diane
  • Field, Ray
  • Pettman, Gary
  • Smales, C. Mark

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