Rapid Process Development for High Yield Plasmid DNA Fed-batch Fermentation

To commercialize DNA medicines, industrial plasmid DNA manufacturing processes that meet the quality, economy, and scale requirements projected for future products are needed. We have developed cell bank and fermentation process unit operation innovations that reduce plasmid-mediated metabolic burden, enabling improved upstream production of optimal plasmids to 2.6 g/L. Application of these processes also facilitated production of otherwise unstable direct repeat containing vectors in standard E. coli host strain DH5α, eliminating the need for specialized stabilizing strains. Fermentation yields with low yield plasmids also were improved up to three-fold by using a simple fermentation process development method requiring only one to two fermentations.