Enhanced plasmid DNA manufacturing using enGenes eXcess technology and an optimized downstream platform

The growing demand for plasmid DNA (pDNA), driven by gene therapies and mRNA vaccines, continues to outpace manufacturing capacity. Current Escherichia coli (E.coli)-based production processes are limited by low strain productivity, complex downstream workflows, high impurity burden, and the sensitivity of alkaline lysis, resulting in scalability challenges and high cost of goods.

This study evaluates the combined impact of strain selection (DH1 vs. e^Xcess™) and process design (conventional vs. optimized lysis and downstream workflow) on plasmid yield, recovery, and overall process efficiency.

The e^Xcess™ strain achieved 7 times higher specific yield (36 vs. 4 µg mL^-1) and >6 times higher volumetric titer (1.17 vs. 0.19 g L^-1 supercoiled pDNA), while maintaining >90% supercoiled purity. The optimized downstream process enabled comparable overall recovery with improved impurity removal and eliminated multiple unit operations, including ultra-/diafiltration (UF/DF), resulting in a reduced total processing time.

Overall, the combination of a high-performance production strain with a streamlined downstream process significantly increases plasmid yield while reducing process complexity, providing a scalable and cost-effective solution for pDNA manufacturing.