\documentclass{article} \usepackage[top=1in,left=1in,right=1in,bottom=1in]{geometry} \usepackage{setspace} \pagestyle{empty} \begin{document} \begin{center} \LARGE{\textbf{Optimizing T Cell Manufacturing and Quality Using Functionalized Degradable Microscaffolds}} \Large \bigskip Nathan J. Dwarshuis \bigskip 165 pages \bigskip Directed by Krishnendu Roy \end{center} \large \doublespacing{} Adoptive cell therapy using chimeric antigen receptor (CAR) T cells have shown promise in treating cancer, but manufacturing large numbers of high quality cells remains challenging. Currently approved T cell expansion technologies involve anti-CD3 and anti-CD28 antibodies, usually mounted on magnetic beads. This method fails to recapitulate many key signals found \textit{in vivo} and is also heavily licensed by a few companies, limiting its long-term usefulness to manufactures and clinicians. Furthermore, highly potent, anti-tumor T cells are generally less-differentiated subtypes such as central memory and stem memory T cells. Despite this understanding, little has been done to optimize T cell expansion for generating these subtypes, including measurement and feedback control strategies that are necessary for any modern manufacturing process. The goal of this dissertation was to develop a microcarrier-based degradable microscaffold (DMS) T cell expansion system and determine biologically-meaningful critical quality attitudes and critical process parameters that could be used to optimize for highly-potent T cells. We developed and characterized the DMS system, including quality control steps. We also demonstrated the feasibility of expanding high-quality T cells. We used Design of Experiments methodology to optimize the DMS platform, and we developed a computational pipeline to identify and model the effects of measurable critical quality attributes and critical process parameters on the final product. Finally, we demonstrated the effectiveness of the DMS platform \textit{in vivo}. This thesis lays the groundwork for a novel T cell expansion method which can be utilized at scale for clinical trials and beyond. \end{document}