Previous small-scale methods for plasmid DNA (pDNA) purification fail to meet theindustry’s demand for sufficient quantities. Greater volumes of bacterial lysates are a consequence of larger volumetric fermentations and traditional large-scale down-stream purification processes have some disadvantages and limitations. The market is believed to continue to expand, hence the need for efficient, cost-effective, andscalable purification processes becomes apparent. A crucial trade-off exists between pDNA yield and purity, necessitating careful consideration in chromatographic pu-rification steps. Each step enhances purity while likely sacrificing yield. In order to achieve a higher degree of pDNA yield, optimal purification entails a single chro-matographic step, specifically anion-exchange chromatography (AEX) in combina-tion with filtration. Alternatively, a two-step purification approach involving AEX followed by hydrophobic interaction chromatography (HIC) is recommended to elim-inate complementary impurities and achieve a high level of purity. Furthermore, the utilization of monolithic chromatographic supports is suggested to facilitate the sug-gested purification strategies. This is due to monoliths promoting higher binding capacities, ensuring robust and consistent results even at high flow rates.