712. Challenges of Production of NanoPlasmid with Large Gene insert using the HyperGRO™ Fermentation Process

Abstract

Plasmid based DNA vaccines are emerging as a promising alternative to traditional vaccines due to several advantages. One of these advantages is faster production of DNA plasmids using E.coli host cells. However, production of plasmids containing large inserts coding for antigen may have various challenges. Bacterial cell machinery may not be able to produce high cell growth during fermentation due to a large gene insert in the plasmid. A Nanoplasmid™ is a plasmid backbone designed by our collaborators, which contains reduced bacterial sequence and produces a smaller plasmid with the gene insert. With the years of expertise in Plasmid production, our company has successfully implemented the HyperGRO™ Fermentation process (an antibiotic-free, fed-batch system) for Nanoplasmid production. Plasmid pNano1, a Nanoplasmid™ with a gene insert of 3762bp, was produced using a modified HyperGRO™process. The fermentation process was successfully scaled up to 10L scale with high end cell density (OD600) of 90.1 and volumetric yield of 0.696g/L.