The demand for plasmid DNA has increased vastly in response to rapid advances in its use in gene therapy and vaccines. These therapies are based on the same principle, i.e. the introduction of nucleic acids in human/non-human cells receptor to restore, cancel, enhance or introduce a biochemical function. Naked plasmid DNA as a vector has attracted a lot of interest since it offers several advantages over a viral vector, especially weak immunogenicity, better safety and easy to manufacture, but low transfection efficacy. Non-viral gene therapy may require considerable amounts (milligram scale) of pharmaceutical-grade pDNA per patient since the efficacy and duration of gene expression is presently relatively low. Reliance on fermentation, which generates large lysate volumes, for producing the needed quantities of pDNA is becoming more widespread. Through optimization of the biological system, growth environment and the growth mode, improvements can be achieved in biomass productivity, plasmid yield, plasmid quality and production costs. The information on large-scale plasmid production is scarce and usually not available to the scientific community. This review summarizes recent patents and patent applications relating to plasmid upstream processing manufacturing, ranging from plasmid design to growth strategies to produce plasmid-bearing E. coli.