Growth medium selection and its economic impact on plasmid DNA production.
Current developments in gene medicine and vaccination studies are utilizing plasmid DNA (pDNA) as the vector. For this reason, there has been an increasing trend towards larger and larger doses of pDNA utilized in human trials: from 100-1000 microg in 2002 to 500-5000 microg in 2005. The increasing demand of pDNA has created the need to revolutionalize current production levels under optimum economy. In this work, different standard media (LB, TB and SOC) for culturing recombinant Escherichia coli DH5alpha harbouring pUC19 were compared to a medium optimised for pDNA production. Lab scale fermentations using the standard media showed that the highest pDNA volumetric and specific yields were for TB (11.4 microg/ml and 6.3 microg/mg dry cell mass respectively) and the lowest was for LB (2.8 microg/ml and 3.3 microg/mg dry cell mass respectively). A fourth medium, PDMR, designed by modifying a stoichiometrically-formulated medium with an optimised carbon source concentration and carbon to nitrogen ratio displayed pDNA volumetric and specific yields of 23.8 microg/ml and 11.2 microg/mg dry cell mass respectively. However, it is the economic advantages of the optimised medium that makes it so attractive. Keeping all variables constant except medium and using LB as a base scenario (100 medium cost [MC] units/mg pDNA), the optimised PDMR medium yielded pDNA at a cost of only 27 MC units/mg pDNA. These results show that greater amounts of pDNA can be obtained more economically with minimal extra effort simply by using a medium optimised for pDNA production.
Danquah
Michael K
MK
Bioengineering Laboratory, Department of Chemical Engineering, Monash University, Clayton campus, Wellington Rd., Victoria 3800, Australia. michael.danquah@eng.monash.edu.au
Forde
Gareth M
GM
D001315:Australia; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D004269:DNA, Bacterial / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development*; D007218:Industrial Microbiology / Q000191:economics / Q000379:methods*; D010957:Plasmids / Q000096:biosynthesis* / Q000191:economics
D004269:DNA, Bacterial