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The application and validation of high-throughput methods in Chinese hamster ovary cell line development

Tamošaitis, Linas (2020) The application and validation of high-throughput methods in Chinese hamster ovary cell line development. 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:83152)

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Language: English

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Abstract

Chinese hamster ovary (CHO) cells are the main production platform for biotherapeutic proteins. In this thesis, two areas were identified and investigated with improving recombinant protein production from CHO cells and cell line development; transcriptomics and technologies for industrial cell line development (CLD).

CLD involves labour and resource intensive cloning out a genetically diverse pool of cells engineered to produce the protein of interest. We sought to analyse a new single cell analysis methodology (Berkley Lights Beacon, BLB) against an industrial ClonePix™ 2 CLD process adapted from FUJIFILM Diosynth Biotechnologies (FDB). We found that there were no statistically significant differences between cell groups generated from the BLB or ClonePix™ 2 processes. Using the Beacon® system, it was possible to predict 3 out of the top 5 producing clones for both Etanercept and Blosozumab. Within the standard ClonePix™ 2 CLD group of cell lines, predictions were most accurate from 24-well plate fed-batch and TubeSpin® batch culture ranks. Further, using the BLB, the time from recovery from transfection to cultures that were ambr® ready was reduced from 65 days to 42 days. Based on the findings of this research it is proposed that the Beacon® is an attractive and powerful new tool in industrial cell line development efforts. To the authors knowledge, this is the first in depth work validating a next-generation CLD process in such detail.

Item Type: Thesis (Doctor of Philosophy (PhD))
Thesis advisor: Smales, Christopher Mark
Uncontrolled keywords: CHO, CLD, cell line development, chinese hamster ovary, siRNA, ClonePix, Beacon, transcriptomics
Subjects: Q Science > QR Microbiology
Divisions: Divisions > Division of Natural Sciences > School of Biosciences
SWORD Depositor: System Moodle
Depositing User: System Moodle
Date Deposited: 28 Sep 2020 08:10 UTC
Last Modified: 29 Sep 2020 10:31 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/83152 (The current URI for this page, for reference purposes)
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