Automated process development for enhanced pDNA production in Vibrio natriegens

As recent reviews highlight, Vibrio natriegens is gaining recognition as a promising chassis for biomanufacturing due to its exceptionally fast growth and high biosynthetic capacity (Lima et al. 2024; Thoma and Blombach 2021; Weinstock et al. 2016). One example is its potential to act as pDNA production host that may be even more effective than Escherichia coli. Given the growing importance of pDNA as a precursor for mRNA and DNA vaccines, exploring alternative hosts with higher productivity and shorter process times is essential. 

The genetic and physiological similarity to E. coli, facilitates the utilization of many established molecular biology and cultivation protocols for the novel organism. However, optimizing experimental procedures to fully harness its potential remains an open challenge, particularly in media formulation. The high number of possible combinations makes manual process development seems unfavorable. We present a high-throughput approach to systematically optimize media composition for V. natriegens in a robotic platform. Using a parallelized minibioreactor system, we accelerate process development by rapidly identifying optimal conditions for growth and productivity in a pDNA formation process. These efforts will pave the way for harnessing V. natriegen’s potential, offering a promising alternative to conventional E. coli-based production systems.