Format

Send to

Choose Destination
Vaccine. 2014 May 19;32(24):2847-50. doi: 10.1016/j.vaccine.2014.02.023. Epub 2014 Mar 2.

Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges.

Author information

1
MIT-Portugal Program, Portugal; Department of Bioengineering, Instituto Superior Técnico (IST), Lisbon, Portugal; IBB-Institute for Biotechnology and Bioengineering, Center for Biological and Chemical Engineering, IST, Lisbon, Portugal.
2
MIT-Portugal Program, Portugal; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.
3
MIT-Portugal Program, Portugal; Department of Bioengineering, Instituto Superior Técnico (IST), Lisbon, Portugal; IBB-Institute for Biotechnology and Bioengineering, Center for Biological and Chemical Engineering, IST, Lisbon, Portugal. Electronic address: miguelprazeres@ist.utl.pt.

Abstract

Plasmid-based vaccines and therapeutics have been making their way into the clinic in the last years. The existence of cost-effective manufacturing processes capable of delivering high amounts of high-quality plasmid DNA (pDNA) is essential to generate enough material for trials and support future commercialization. However, the development of pDNA manufacturing processes is often hampered by difficulties in predicting process scale performance of Escherichia coli cultivation on the basis of results obtained at lab scale. This paper reports on the differences observed in pDNA production when using shake flask and bench-scale bioreactor cultivation of E. coli strains MG1655ΔendAΔrecA and DH5α in complex media with 20 g/L of glucose. MG1655ΔendAΔrecA produced 5-fold more pDNA (9.8 mg/g DCW) in bioreactor than in shake flask (1.9 mg/g DCW) and DH5α produced 4-fold more pDNA (8 mg/g DCW) in bioreactor than in shake flask (2 mg/g DCW). Accumulation of acetate was also significant in shake flasks but not in bioreactors, a fact that was attributed to a lack of control of pH.

KEYWORDS:

DNA vaccine; Escherichia coli; Fermentation; Plasmid biopharmaceuticals; Scale-up process; Strain engineering

PMID:
24598722
DOI:
10.1016/j.vaccine.2014.02.023
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center