Fed-batch microbioreactor platform for scale down and analysis of a plasmid DNA production process

The rising costs of bioprocess research and development emphasize the need for high‐throughput, low‐cost alternatives to bench‐scale bioreactors for process development. In particular, there is a need for platforms that can go beyond simple batch growth of the organism of interest to include more advanced monitoring, control, and operation schemes such as fed‐batch or continuous. We have developed a 1‐mL microbioreactor capable of monitoring and control of dissolved oxygen, pH, and temperature. Optical density can also be measured online for continuous monitoring of cell growth. To test our microbioreactor platform, we used production of a plasmid DNA vaccine vector (pVAX1‐GFP) in Escherichia coli via a fed‐batch temperature‐inducible process as a model system. We demonstrated that our platform can accurately predict growth, glycerol and acetate concentrations, as well as plasmid copy number and quality obtained in a bench‐scale bioreactor. The predictive abilities of the micro‐scale system were robust over a range of feed rates as long as key process parameters, such as dissolved oxygen, were kept constant across scales. We have highlighted plasmid DNA production as a potential application for our microbioreactor, but the device has broad utility for microbial process development in other industries as well. Biotechnol. Bioeng. 2012; 109:1976–1986. © 2012 Wiley Periodicals, Inc. The authors have developed a microbioreactor with a 1‐mL working volume that is capable of fed‐batch operation; control of dissolved oxygen, pH, and temperature; as well as continuous monitoring of cell growth. In this work, The authors demonstrated that the micro‐scale device can accurately mimic a complex, bench‐scale plasmid DNA production process. They also identified key process parameters required for consistency of results across scales.