Novel Workflow for Activation and Expansion of T Cells Using Microbubble Technology

Abstract Chimeric antigen receptor (CAR) T cell therapy was developed to address the significant hurdles to cancer treatment. In the US, one in four deaths is due to cancer, with treatment costs totaling over $200 billion a year. Traditional treatments, such as chemotherapy and radiation therapy have serious harmful side effects, often with limited success. Despite the immense hurdles to producing a cost effective, efficient, and scalable workflow for manufacturing CAR T cells, thousands s of CAR T cell therapy clinical trials have been initiated around the world. Traditional CAR T cell workflows often result in T cell exhaustion, limiting the effectiveness of the cells produced. At Akadeum, we set out to create a simple workflow to improve researchers’ ability to develop custom cell therapies. Using Akadeum’s microbubbles, we developed a process that combines selection, activation, and expansion into a single workflow. Because of the natural buoyancy of the microbubbles, activation and expansion take place at the top of the culture. As the T cells divide, daughter cells fall to the bottom of the well away from the activation signals. This prevents overstimulation and the resulting exhaustion phenotype that occurs with the industry-standard activation methods (25–30% PD-1+ with Akadeum method compared to >90% with traditional workflow). This method effectively activates T cells, with significant upregulation of standard activation markers, greater than 20-fold expansion, and desirable effector cell ratios. These cells are also highly transducible. Taken together, these data show that Akadeum’s microbubble-based workflow provides all of the critical aspects of CAR T cell generation while eliminating many of the most significant hurdles.