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Title: High-throughput platform development for Chinese Hamster Ovary (CHO) cell culture performance enhancement using small molecules
Author: Kalsi, Devika
ISNI:       0000 0004 7651 3515
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2018
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While g L⁻¹ quantities of recombinant protein are common in Chinese Hamster Ovary (CHO) cell factories, the rise of more complex products presents novel upstream manufacturing challenges. Bioactive small molecule enhancers (SMEs) present opportunities for targeted CHO culture performance improvements. Here we present the development of a high-throughput (HT) screening technology for the identification of SMEs and their combinations for improved bioprocess. Firstly, we describe the development of the HT screening platform. A miniaturised, shaking culture methodology was developed, enabling rapid assessment of 96 cultures simultaneously. Growth and production performance was similar to shake flask cultures, demonstrating scalability. HT growth, viability and titer measurement technologies were established. Secondly, a large suite of SMEs was evaluated for the purposes of CHO cell biopharmaceutical production. 43 SMEs were assessed rapidly using the HT culturing and analytical platforms. Molecules that improved cellular growth or production in a stable production format were used to inform combinatorial designs to determine interactions for amplifying culture performance. This work would form the foundation of a commercial screening tool, wherein a deep well plate product pre-coated with selected enhancer concentrations and combinations would be available for testing with cell lines and products of choice. The product along with statistical modelling tools like Design of Experiments methodologies would advise chemical supplementation strategies to create bespoke media for enhanced bioprocess. Finally, we demonstrate another use of the HT screening tool for the discovery of novel molecules for biotherapeutic production. Molecules were selected based on structural similarity to established titer enhancing SMEs and evaluated using the HT platform. A novel molecule and associated parent molecule maintained production improvements when scaled-up to shake flasks. Various mechanistic analyses revealed that the molecule acted epigenetically, resulting in higher transcription of the product. Given the age of biosimilars/biobetters, having a competitive edge in terms of speed and cost is crucial. The HT screening tool developed utilises the potential of SMEs as quick, simple and cost-effective methods for improvements in upstream bioprocessing.
Supervisor: James, David Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available