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Segregated growth kinetics ofEscherichia coliDH5α-NH36 in exponential-fed perfusion culture for pDNA vaccine production
- Source :
- Biotechnology and Applied Biochemistry. 62:795-805
- Publication Year :
- 2015
- Publisher :
- Wiley, 2015.
-
Abstract
- The clinical demand of plasmid DNA (pDNA) has been increasing constantly. An exponential-fed perfusion (EFP) culture is a new mode for plasmid production for clinical trials and commercialization. However, the culture conditions may lead to cell filamentation and growth cessation. In this study, the variation of the physiological state and the plasmid contents of Escherichia coli DH5α hosting pVAX1-NH36 in an EFP culture for application as a Leishmaniasis vaccine was investigated. The culture performance was monitored using flow cytometry (FC) and real-time quantitative PCR. The FC studies showed a high viability of cell population and a constant distribution of complexity and size. A high homogeneity of pDNA (>95 % of supercoiled) was obtained, which might be attributed to a better culture environment. The obtained plasmid specific and volumetric yields of 1.8 mg/g dcw and 36.5 mg/L represent typical values for laboratory-scale plasmid production in a defined medium. A segregated kinetic model of the perfusion system was developed and fitted to the experimental data (R(2) > 0.96). A practical conclusion of this work is that a space-time yield analysis of a bioprocess requires a viability evaluation. This new strategy of culture operation might help in the efficient production of pDNA for therapeutic use.
- Subjects :
- education.field_of_study
medicine.diagnostic_test
Process Chemistry and Technology
Population
Biomedical Engineering
Bioengineering
General Medicine
Biology
medicine.disease_cause
Applied Microbiology and Biotechnology
Molecular biology
Flow cytometry
Chemically defined medium
Perfusion Culture
Plasmid
Drug Discovery
medicine
Molecular Medicine
Distribution (pharmacology)
Bioprocess
education
Escherichia coli
Biotechnology
Subjects
Details
- ISSN :
- 08854513
- Volume :
- 62
- Database :
- OpenAIRE
- Journal :
- Biotechnology and Applied Biochemistry
- Accession number :
- edsair.doi...........805d6cfef164a56ab8df5dc06120fa91