Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking
Bioprocessing has been transitioning from batch to continuous processes. As a result, a considerable amount of resource was dedicated to optimising strategies for continuous production. However, the focus has been on developing a suitable and scalable perfusion strategy with little attention given t...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1479633/full |
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| author | Marie Dorn Christine Ferng Kerensa Klottrup-Rees Kenneth Lee Martina Micheletti |
| author_facet | Marie Dorn Christine Ferng Kerensa Klottrup-Rees Kenneth Lee Martina Micheletti |
| author_sort | Marie Dorn |
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| description | Bioprocessing has been transitioning from batch to continuous processes. As a result, a considerable amount of resource was dedicated to optimising strategies for continuous production. However, the focus has been on developing a suitable and scalable perfusion strategy with little attention given to the selection of optimal cell clones. Cell line development and lead clone selection are critical to bioprocess development. The screening and selection process is typically performed in stages. Microwell plates (MWP) are used to narrow down the number of clone candidates, which will undergo further selective screening in progressively larger small-scale bioreactors (12 mL–3 L) to identify the top clone for GMP production. Perfusion mode is typically applied at bench-scale for optimisation purposes, while process development and cell clone screening studies at mL-scale still commonly use fed-batch methods. The change of operation mode from bolus feeding to perfusion with a regular exchange of medium, leads to questions regarding the reliability and fit of initial clone selection. Is the early-stage clone ranking impacted by the discrepancy in the operation mode, and does this potentially result in the exclusion of cell clones suitable for perfusion processes? To address this question, we evaluated various CHO cell clones expressing two antibody products using MWP methodologies in fed-batch and semi-perfusion mode. We assessed growth, metabolic, and productivity performance, and ranked cell clones using two different strategies. The first strategy evaluated clones based on a single parameter: the cell-specific productivity (qP). The second considered a collection of multiple parameters using the metric of the Manufacturability index (MICL). Both ranking strategies showed an impact of operation mode and perfusion rate on the clone ranking. Notably, depending on the chosen operation mode, different sets of candidate clones might have been selected for further, more extensive screening. Additionally, we evaluated the reproducibility of our results demonstrating consistency in cell clone growth performance and ranking. |
| format | Article |
| id | doaj-art-777e4d66ace541458cc03bae2562a7c9 |
| institution | Kabale University |
| issn | 2296-4185 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj-art-777e4d66ace541458cc03bae2562a7c92025-01-20T07:19:39ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852025-01-011210.3389/fbioe.2024.14796331479633Cell clone selection—impact of operation modes and medium exchange strategies on clone rankingMarie Dorn0Christine Ferng1Kerensa Klottrup-Rees2Kenneth Lee3Martina Micheletti4Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, London, United KingdomBioProcess Technologies and Engineering, Biopharmaceutical Developments, AstraZeneca, Gaithersburg, MD, United StatesCell Culture and Fermentation Sciences, Biopharmaceutical Development, AstraZeneca, Cambridge, United KingdomBioProcess Technologies and Engineering, Biopharmaceutical Developments, AstraZeneca, Gaithersburg, MD, United StatesAdvanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, London, United KingdomBioprocessing has been transitioning from batch to continuous processes. As a result, a considerable amount of resource was dedicated to optimising strategies for continuous production. However, the focus has been on developing a suitable and scalable perfusion strategy with little attention given to the selection of optimal cell clones. Cell line development and lead clone selection are critical to bioprocess development. The screening and selection process is typically performed in stages. Microwell plates (MWP) are used to narrow down the number of clone candidates, which will undergo further selective screening in progressively larger small-scale bioreactors (12 mL–3 L) to identify the top clone for GMP production. Perfusion mode is typically applied at bench-scale for optimisation purposes, while process development and cell clone screening studies at mL-scale still commonly use fed-batch methods. The change of operation mode from bolus feeding to perfusion with a regular exchange of medium, leads to questions regarding the reliability and fit of initial clone selection. Is the early-stage clone ranking impacted by the discrepancy in the operation mode, and does this potentially result in the exclusion of cell clones suitable for perfusion processes? To address this question, we evaluated various CHO cell clones expressing two antibody products using MWP methodologies in fed-batch and semi-perfusion mode. We assessed growth, metabolic, and productivity performance, and ranked cell clones using two different strategies. The first strategy evaluated clones based on a single parameter: the cell-specific productivity (qP). The second considered a collection of multiple parameters using the metric of the Manufacturability index (MICL). Both ranking strategies showed an impact of operation mode and perfusion rate on the clone ranking. Notably, depending on the chosen operation mode, different sets of candidate clones might have been selected for further, more extensive screening. Additionally, we evaluated the reproducibility of our results demonstrating consistency in cell clone growth performance and ranking.https://www.frontiersin.org/articles/10.3389/fbioe.2024.1479633/fullperfusionfed-batchsmall-scalemicrowell platehigh-throughputcell clone screening |
| spellingShingle | Marie Dorn Christine Ferng Kerensa Klottrup-Rees Kenneth Lee Martina Micheletti Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking Frontiers in Bioengineering and Biotechnology perfusion fed-batch small-scale microwell plate high-throughput cell clone screening |
| title | Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking |
| title_full | Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking |
| title_fullStr | Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking |
| title_full_unstemmed | Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking |
| title_short | Cell clone selection—impact of operation modes and medium exchange strategies on clone ranking |
| title_sort | cell clone selection impact of operation modes and medium exchange strategies on clone ranking |
| topic | perfusion fed-batch small-scale microwell plate high-throughput cell clone screening |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2024.1479633/full |
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