Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control
Abstract Unfavorable process conditions lead to adverse cultivation states, limited cell growth and thus hamper biotherapeutic protein production. Oxygen deficiency or hyperosmolality are among the most critical process conditions and therefore require continuous monitoring. We established a novel s...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-02-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-06272-x |
| _version_ | 1818973754781335552 |
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| author | Nikolas Zeh Melina Bräuer Nadja Raab René Handrick Kerstin Otte |
| author_facet | Nikolas Zeh Melina Bräuer Nadja Raab René Handrick Kerstin Otte |
| author_sort | Nikolas Zeh |
| collection | DOAJ |
| description | Abstract Unfavorable process conditions lead to adverse cultivation states, limited cell growth and thus hamper biotherapeutic protein production. Oxygen deficiency or hyperosmolality are among the most critical process conditions and therefore require continuous monitoring. We established a novel sensor CHO cell line with the ability to automatically sense and report unwanted process conditions by the expression of destabilized fluorescent proteins. To this end, an inducible real-time system to detect hypoxia by hypoxia response elements (HREs) of vascular endothelial growth factor (VEGF) origin reporting limitations by the expression of destabilized green fluorescent protein (GFP) was created. Additionally, we established a technique for observing hyperosmolality by exploiting osmotic response elements (OREs) for the expression of unstable blue fluorescent protein (BFP, FKBP-BFP), enabling the simultaneous automated supervision of two bioprocess parameters by using a dual sensor CHO cell line transfected with a multiplexable monitoring system. We finally also provided a fully automated in-line fluorescence microscopy-based setup to observe CHO cells and their response to varying culture conditions. In summary, we created the first CHO cell line, reporting unfavorable process parameters to the operator, and provided a novel and promising sensor technology accelerating the implementation of the process analytical technology (PAT) initiative by innovative solutions. |
| first_indexed | 2024-12-20T15:29:12Z |
| format | Article |
| id | doaj.art-c9097d764281471ebcd8700ada13a90f |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T15:29:12Z |
| publishDate | 2022-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-c9097d764281471ebcd8700ada13a90f2022-12-21T19:35:40ZengNature PortfolioScientific Reports2045-23222022-02-0112111110.1038/s41598-022-06272-xExploring synthetic biology for the development of a sensor cell line for automated bioprocess controlNikolas Zeh0Melina Bräuer1Nadja Raab2René Handrick3Kerstin Otte4Institute of Applied Biotechnology, University of Applied Sciences BiberachInstitute of Applied Biotechnology, University of Applied Sciences BiberachInstitute of Applied Biotechnology, University of Applied Sciences BiberachInstitute of Applied Biotechnology, University of Applied Sciences BiberachInstitute of Applied Biotechnology, University of Applied Sciences BiberachAbstract Unfavorable process conditions lead to adverse cultivation states, limited cell growth and thus hamper biotherapeutic protein production. Oxygen deficiency or hyperosmolality are among the most critical process conditions and therefore require continuous monitoring. We established a novel sensor CHO cell line with the ability to automatically sense and report unwanted process conditions by the expression of destabilized fluorescent proteins. To this end, an inducible real-time system to detect hypoxia by hypoxia response elements (HREs) of vascular endothelial growth factor (VEGF) origin reporting limitations by the expression of destabilized green fluorescent protein (GFP) was created. Additionally, we established a technique for observing hyperosmolality by exploiting osmotic response elements (OREs) for the expression of unstable blue fluorescent protein (BFP, FKBP-BFP), enabling the simultaneous automated supervision of two bioprocess parameters by using a dual sensor CHO cell line transfected with a multiplexable monitoring system. We finally also provided a fully automated in-line fluorescence microscopy-based setup to observe CHO cells and their response to varying culture conditions. In summary, we created the first CHO cell line, reporting unfavorable process parameters to the operator, and provided a novel and promising sensor technology accelerating the implementation of the process analytical technology (PAT) initiative by innovative solutions.https://doi.org/10.1038/s41598-022-06272-x |
| spellingShingle | Nikolas Zeh Melina Bräuer Nadja Raab René Handrick Kerstin Otte Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control Scientific Reports |
| title | Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| title_full | Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| title_fullStr | Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| title_full_unstemmed | Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| title_short | Exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| title_sort | exploring synthetic biology for the development of a sensor cell line for automated bioprocess control |
| url | https://doi.org/10.1038/s41598-022-06272-x |
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