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Development of mL-scale Pseudo-Perfusion Methodologies for High-Throughput Early Phase Development Studies

Tregidgo, Molly; Lucas, Ciara; Dorn, Marie; Martina, Micheletti; (2023) Development of mL-scale Pseudo-Perfusion Methodologies for High-Throughput Early Phase Development Studies. Biochemical Engineering Journal , Article 108906. 10.1016/j.bej.2023.108906. Green open access

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Abstract

Small-scale devices enable rapid development and optimization, and while many exist for fed-batch cultures, there are few comparable devices for high cell density perfusion cultures. To address this gap, pseudo-perfusion methodologies were developed in microwell plates at mL-scale capable of achieving many of the specific characteristics of perfusion culture, including high cell density, cell retention and increased productivity. Pseudo-perfusion was achieved via sedimentation or centrifugation of 24 microwell plates prior to media exchange 1–2 times daily, generating separation efficiencies higher than 90%. Media exchanges commenced on day 3 and achieved perfusion rates of 0.5–1.8 vessel volumes per day (VVD). Pseudo-perfusion methodologies resulted in maximum viable cell densities (VCDs) of up to 42 × 106 cells mL-1, 4.2-fold greater than fed-batch cultures. Volumetric productivities increased by 1.9-fold, generating industrially relevant productivities. Similar performance was observed between sedimentation and centrifugation methodologies, with minor deviations attributed to elongated manipulation times and lower packed cell density prior to separation when sedimentation was used. The microwell plate (MWP) experiments were validated at the 5 L scale and showed comparability in metabolite and growth profiles. The work presents the development of a robust tool for high-throughput development studies which is sensitive to changes in media composition and exchange rate, which could be reliably used for initial screening of high cell density perfusion cultures.

Type: Article
Title: Development of mL-scale Pseudo-Perfusion Methodologies for High-Throughput Early Phase Development Studies
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.bej.2023.108906
Publisher version: https://doi.org/10.1016/j.bej.2023.108906
Language: English
Additional information: © 2023 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: CHO cells, Microscale, Perfusion culture, Monoclonal antibodies, High throughput
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/10167967
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