Generic plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch fermentation processes
| Autor: | Aaron E. Carnes, Sarah Langtry, Clague P. Hodgson, James A. Williams, Jeremy Luke, Sheryl Anderson |
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| Rok vydání: | 2009 |
| Předmět: |
DNA
Bacterial biology business.industry Hemagglutinin Glycoproteins Influenza Virus Bioengineering Computational biology biology.organism_classification Applied Microbiology and Biotechnology Unit operation Article Recombinant Proteins DNA vaccination Biotechnology Plasmid Antigen Escherichia coli Vaccines DNA Fermentation Cell bank business Gene Bacteria Plasmids |
| Zdroj: | Biotechnology and Bioengineering. 103:1129-1143 |
| ISSN: | 1097-0290 0006-3592 |
| DOI: | 10.1002/bit.22347 |
| Popis: | DNA vaccines have tremendous potential for rapid deployment in pandemic applications, wherein a new antigen is ‘plugged’ into a validated vector, and rapidly produced in a validated, fermentation - purification process. For this application, it is essential that the vector and fermentation process function with a variety of different antigen genes. However, many antigen genes are unpredictably ‘toxic’ or otherwise low yielding in standard fermentation processes. We report cell bank and fermentation process unit operation innovations that reduce plasmid-mediated metabolic burden, enabling successful production of previously known toxic influenza hemagglutinin antigen genes. These processes, combined with vector backbone modifications, doubled fermentation productivity compared to existing high copy vectors, such as pVAX1 and gWIZ, resulting in high plasmid yields (up to 2220 mg/L, 5% of total dry cell weight) even with previously identified toxic or poor producing inserts. |
| Databáze: | OpenAIRE |
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