Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium
The recognition of the potential efficacy of plasmid DNA (pDNA) molecules as vectors in the treatment and prevention of emerging diseases has birthed the confidence to combat global pandemics. This is due to the close-to-zero safety concern associated with pDNA vectors compared to viral vectors in c...
| Main Authors: | , |
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| Format: | Journal Article |
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Korean Society for Biotechnology and Bioengineering
2008
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| Online Access: | http://hdl.handle.net/20.500.11937/10972 |
| _version_ | 1848747680184926208 |
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| author | Danquah, Michael Forde, G. |
| author_facet | Danquah, Michael Forde, G. |
| author_sort | Danquah, Michael |
| building | Curtin Institutional Repository |
| collection | Online Access |
| description | The recognition of the potential efficacy of plasmid DNA (pDNA) molecules as vectors in the treatment and prevention of emerging diseases has birthed the confidence to combat global pandemics. This is due to the close-to-zero safety concern associated with pDNA vectors compared to viral vectors in cell transfection and targeting. Considerable attention has been paid to the potential of pDNA vectors but comparatively less thought has been given to the practical challenges in producing large quantities to meet current rising demands. A pilot-scale fermentation scheme was developed by employing a stoichiometrically-designed growth medium whose exceptional plasmid yield performance was attested in a shake flask environment for pUC19 and pEGFP-N1 transformed into E. coliDH5a and E. coliJM109, respectively. Batch fermentation of E. coliDH5a-pUC19 employing the stoichiometric medium displayed a maximum plasmid volumetric and specific yield of 62.6 mg/L and 17.1 mg/g (mg plasmid/g dry cell weight), respectively. Fed-batch fermentation of E. coliDH5a-pUC19 on a glycerol substrate demonstrated one of the highest ever reported pilot-scale plasmid specific yield of 48.98 mg/g and a volumetric yield of 0.53 g/L. The attainment of high plasmid specific yields constitutes a decrease in plasmid manufacturing cost and enhances the effectiveness of downstream processes by reducing the proportion of intracellular impurities. The effect of step-rise temperature induction was also considered to maximize ColE1-origin plasmid replication. © KSBB. |
| first_indexed | 2025-11-14T06:53:00Z |
| format | Journal Article |
| id | curtin-20.500.11937-10972 |
| institution | Curtin University Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-14T06:53:00Z |
| publishDate | 2008 |
| publisher | Korean Society for Biotechnology and Bioengineering |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | curtin-20.500.11937-109722017-09-13T14:55:44Z Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium Danquah, Michael Forde, G. The recognition of the potential efficacy of plasmid DNA (pDNA) molecules as vectors in the treatment and prevention of emerging diseases has birthed the confidence to combat global pandemics. This is due to the close-to-zero safety concern associated with pDNA vectors compared to viral vectors in cell transfection and targeting. Considerable attention has been paid to the potential of pDNA vectors but comparatively less thought has been given to the practical challenges in producing large quantities to meet current rising demands. A pilot-scale fermentation scheme was developed by employing a stoichiometrically-designed growth medium whose exceptional plasmid yield performance was attested in a shake flask environment for pUC19 and pEGFP-N1 transformed into E. coliDH5a and E. coliJM109, respectively. Batch fermentation of E. coliDH5a-pUC19 employing the stoichiometric medium displayed a maximum plasmid volumetric and specific yield of 62.6 mg/L and 17.1 mg/g (mg plasmid/g dry cell weight), respectively. Fed-batch fermentation of E. coliDH5a-pUC19 on a glycerol substrate demonstrated one of the highest ever reported pilot-scale plasmid specific yield of 48.98 mg/g and a volumetric yield of 0.53 g/L. The attainment of high plasmid specific yields constitutes a decrease in plasmid manufacturing cost and enhances the effectiveness of downstream processes by reducing the proportion of intracellular impurities. The effect of step-rise temperature induction was also considered to maximize ColE1-origin plasmid replication. © KSBB. 2008 Journal Article http://hdl.handle.net/20.500.11937/10972 10.1007/s12257-007-0080-2 Korean Society for Biotechnology and Bioengineering restricted |
| spellingShingle | Danquah, Michael Forde, G. Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title | Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title_full | Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title_fullStr | Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title_full_unstemmed | Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title_short | Development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| title_sort | development of a pilot-scale bacterial fermentation for plasmid-based biopharmaceutical production using a stoichiometric medium |
| url | http://hdl.handle.net/20.500.11937/10972 |