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Integrating upstream and downstream process development strategies for mammalian cell derived therapeutic antibodies

Wilson, Louisa Jane; (2020) Integrating upstream and downstream process development strategies for mammalian cell derived therapeutic antibodies. Doctoral thesis (Ph.D), UCL (University College London). Green open access

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Abstract

Recent improvements in volumetric antibody productivity (often in excess of 5 g/L) have been achieved by advances in cell lines and upstream processing, but often lead to harvest material becoming more difficult to recover. These intensified upstream operations require a renewed prioritisation of the integration of upstream and downstream process development to ensure product purification issues are taken into consideration, to avoid extensive and expensive clearance strategies downstream. Here, it was demonstrated that changes to upstream process parameters at the bioreactor stage of monoclonal antibody production affect product quantity and quality. Culture pH, temperature and seed density setpoints leading to high titre are commonly also linked to higher post-protein A HCP levels, reduced monomer percentages and increased percentages of undesirable glycan structures. To predict post-protein A product quality, several potential indicators that can be measured in harvest material (prior to using expensive purification resources) were explored, including culture viability and osmolality, revealing unexpectedly that culture viability could not be used for such a purpose, but that osmolality has the potential to be used as a product quality indicator. The impact of culture duration on product quality was also investigated and it was shown that as cultivation progressed and antibody titre increased, product quality declined, in one case due to post-protein A HCP levels increasing by 75% from day 14 to day 17 of culture. HCP identification by mass spectrometry was applied to this system to provide insights into cellular behaviour and HCP carryover during protein A purification. It showed increases in several classes of post-protein A HCPs (e.g. stress response proteins) as the culture progressed, particularly on days 15 and 17 of culture which were associated with significant increases in total HCP levels. This provides a new level of insight into HCPs that are retained during mAb purification which may be used to aide process development strategies.

Type: Thesis (Doctoral)
Qualification: Ph.D
Title: Integrating upstream and downstream process development strategies for mammalian cell derived therapeutic antibodies
Event: UCL (University College London)
Open access status: An open access version is available from UCL Discovery
Language: English
Additional information: Copyright © The Author 2020. Original content in this thesis is licensed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) Licence (https://creativecommons.org/licenses/by/4.0/). Any third-party copyright material present remains the property of its respective owner(s) and is licensed under its existing terms. Access may initially be restricted at the author’s request.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10104331
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