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Investigation of Translational Reprogramming during Transient and Stable Expression of Monoclonal Antibodies in Chinese Hamster Ovary Cells

Godfrey, Charlotte (2017) Investigation of Translational Reprogramming during Transient and Stable Expression of Monoclonal Antibodies in Chinese Hamster Ovary Cells. Doctor of Philosophy (PhD) thesis, University of Kent,. (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided) (KAR id:65774)

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Abstract

Translational reprogramming and mRNA translation e ciency greatly in uence global protein synthesis, cell proliferation and growth; important parameters in de ning recombinant protein expression yields. Polysome pro ling is a widely-used technique to analyse mRNA transla- tion and its e ciency that provides a snapshot of ribosomes loaded on mRNA transcripts at any particular time. A higher number of polysomes present on a given mRNA suggests that the mRNA is being more heavily translated than those mRNAs with few ribosomes. Fur- ther, a large pool of sub-polysomes (40S, 60S and 80S) compared to polysomes in a sample suggests low translational activity. Here, polysome pro ling has been applied to investigate translational reprogramming in multiple recombinant monoclonal antibody (mAb)-producing Chinese hamster ovary (CHO) cell lines, and to determine how reprogramming re ects the ability of such cells to proliferate and make recombinant proteins in stable and transient mAb expression systems, in batch and fed-batch culture mode. The impact of culture temperature on the polysome pro le and hence on reprogramming was also investigated in transient studies.

The ndings described in this thesis therefore reveal insights into the mechanisms involved in translational regulation and reprogramming in CHO cells during bioprocessing. These can be utilised for further improvement via targeted cell engineering strategies, cell line screen- ing approaches or modi cation of recombinant transcripts for enhanced industrial host and recombinant cell lines.

Item Type: Thesis (Doctor of Philosophy (PhD))
Thesis advisor: Smales, Christopher Mark
Uncontrolled keywords: CHO, Polysome profiling, Monoclonal antibodies, Translation, Reprogramming, Recombinant protein
Divisions: Faculties > Sciences > School of Biosciences
SWORD Depositor: System Moodle
Depositing User: System Moodle
Date Deposited: 24 Jan 2018 11:11 UTC
Last Modified: 29 May 2019 20:11 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/65774 (The current URI for this page, for reference purposes)
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