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Biotechnol Bioeng. 2009 Aug 15;103(6):1129-43. doi: 10.1002/bit.22347.

Generic plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch fermentation processes.

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1
Nature Technology Corporation, 4701 Innovation Drive, Lincoln, Nebraska 68521, USA.

Abstract

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 2,220 mg/L, 5% of total dry cell weight) even with previously identified toxic or poor producing inserts.

PMID:
19408315
PMCID:
PMC2735187
DOI:
10.1002/bit.22347
[Indexed for MEDLINE]
Free PMC Article
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