PhD Dissertation Defense: Chad Sundberg

Cell-free systems have recently gained prominence as biomanufacturing technologies due to their rapid lead times and versatility in producing a diverse array of products. However, the identification of critical process parameters and mechanisms that drive economically viable yields remains challenging, thereby limiting their commercial applicability. The objective of my doctoral research was to develop specialized bioreactors for cell-free synthesis reactions with the aims of 1) enhancing reaction productivity and control, and 2) facilitating the identification of biochemical mechanisms that underpin economic viability, productivity, and scalability. These efforts resulted in 1) significant productivity increases relative to industry collaborators and published metrics, 2) a reduction in reaction durations from approximately 20 hours to 8 and 4 hours respectively and 3) and significant increases in grams of product yield per dollar of reagents input. More significantly, our reactors have enabled the collection of real-time process data at the milliliter scale to investigate energy metabolism, metabolic flux, and product synthesis rates, which we anticipate will inform process control, design, and development for commercial applications such as personalized medicines, forward-deployable biomanufacturing platforms, and sustainable commodity chemical synthesis methods.