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Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges.
Vaccine. 2014 May 19; 32(24):2847-50.V

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.

Authors+Show Affiliations

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.MIT-Portugal Program, Portugal; Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.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.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.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24598722

Citation

Gonçalves, Geisa A L., et al. "Engineering of Escherichia Coli Strains for Plasmid Biopharmaceutical Production: Scale-up Challenges." Vaccine, vol. 32, no. 24, 2014, pp. 2847-50.
Gonçalves GA, Prather KL, Monteiro GA, et al. Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges. Vaccine. 2014;32(24):2847-50.
Gonçalves, G. A., Prather, K. L., Monteiro, G. A., & Prazeres, D. M. (2014). Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges. Vaccine, 32(24), 2847-50. https://doi.org/10.1016/j.vaccine.2014.02.023
Gonçalves GA, et al. Engineering of Escherichia Coli Strains for Plasmid Biopharmaceutical Production: Scale-up Challenges. Vaccine. 2014 May 19;32(24):2847-50. PubMed PMID: 24598722.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges. AU - Gonçalves,Geisa A L, AU - Prather,Kristala L J, AU - Monteiro,Gabriel A, AU - Prazeres,Duarte M F, Y1 - 2014/03/02/ PY - 2014/3/7/entrez PY - 2014/3/7/pubmed PY - 2014/11/7/medline KW - DNA vaccine KW - Escherichia coli KW - Fermentation KW - Plasmid biopharmaceuticals KW - Scale-up process KW - Strain engineering SP - 2847 EP - 50 JF - Vaccine JO - Vaccine VL - 32 IS - 24 N2 - 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. SN - 1873-2518 UR - https://www.unboundmedicine.com/medline/citation/24598722/Engineering_of_Escherichia_coli_strains_for_plasmid_biopharmaceutical_production:_scale_up_challenges_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0264-410X(14)00196-0 DB - PRIME DP - Unbound Medicine ER -