Large-scale plasmid production for gene therapy presents very specific problems with regards to the reproducibility of process.
Solutions for these problems and others will undoubtedly have an impact on the economics, efficacy, and safety of non-viral
approaches to gene therapy. As advances continue in the field of DNA vaccines, factories capable of producing kilograms of
pDNA per year must be designed.
Scale-up of plasmid pIDKE2 fermentation from a 5-L fermentor to 50-L pilot-scale fermentor was carried out successfully using
P/V constant in a fed-batch process. With this culture procedure, larger amounts of plasmid DNA can be obtained in DH10B cells.
The results from this study may be beneficial to the development of techniques for the fed-batch cultivation of E. coli cells and for the efficient large-scale production of plasmid DNA for therapeutic use in humans.
Odalys Ruiz Hernández* is a principal researcher in upstream process development, Jorge Valdes Hernández is head of the Fermentation Development Department, Willy Frometa Planche is a specialist in upstream process development, Michel Diaz Martínez is a specialist in upstream process development, Daniel Alvares Almiñaque is a specialist in upstream process development, Marta Pupo Peña is a researcher in the Analytical Development Department, Miladys Limonta Fernández is a researcher in downstream development of biomolecules, Dinorah Torres Idahody is the head of the Analytical Development Department, and Eduardo Martínez is head of the Development Division, all at the Center for Genetic Engineering and Biotechnology, Havana, Cuba.* To whom
correspondance should be addressed, email@example.com
1. K.J. Prather et at., Enzyme Microb. Technol.
33, 865–883 (2003).
2. M. Limonta et al., BioPharm Intl.
21 (9), 38–47 (2008).
3. H. Robinson, Clin. Microbiol. Newsletter
23, 17–22 (2000).
4. R. O´Kennedy, J. Ward, and E. Keshavarz. Biotecnol. Appl. Biochem.
37, 83–90 (2003).
5. B. Yakhchali et al., Jrnl. of Sci.
18 (2), 129–133. (2007).
6. A. Carnes, BioProcess Technic. 2–7 (2005).
7. M. Schleef, Biotechnol.
5a, 445–469 (1999).
8. Ch. Kim et al., Biotechnology and Bioprocess Engin.
8, 303–305 (2003).
9. D. Pollard et al., Biotechnol. Bioeng. 96 (2), 307–307 (2007).
10. O. Ruiz et al., BioPharm Intl. 22 (7), 40–45 (2009).
11. Z. Xu et al., Bioproc. Eng.
23, 669–674 (2000).
12. G. Miller, Anal. Chem.
31, 426–428 (1959).
13. H. C. Birnboim and J. Doly, Nucleic Acids Res.
7, 1513–1523. (1979).
14. S. Aiba, A. Humphrey and F. Millis, Biochemical Engineering (Academic Press, NY, 2nd ed., 1973). pp. 17–28, 75–83, 107–113, 133–149, 163–170.
15. M. Diogo et al., J. Gene Med.
3, 577–584 (2001).
16. N. Horn et al., Hum. Gene Therapy. l6, 565–73. (1995).