Design of CAR-T Cell Manufacturing Process

CAR T-cell therapy is at the frontier of personalized immunotherapy. It is a therapy that essentially reprograms a patient’s own T-cells to attack certain blood cancers. A sample of a patient's T cells are collected from the blood, then modified to produce chimeric antigen receptors (CARs) on their surface. When these CAR T cells are reinfused into the patient, the new receptors enable them to latch onto a specific antigen on the patient's tumor cells and kill them, ideally sending the patient into remission and essentially curing their cancer. Currently, CAR T-cell therapy is FDA approved as standard of care for some forms of aggressive, refractory non-Hodgkin lymphoma and for patients with relapsed or refractory acute lymphoblastic leukemia up to age 25. There is a great deal of development occurring to use this therapy in solid tumor cancers, which will bolster the need for large-scale manufacturing processes. This project seeks to develop and optimize a large-scale parallelizable manufacturing process for CAR-T cell therapy. To begin this manufacturing process, whole blood is drawn from a patient and passed through a filter to collect the leukocytes. These leukocytes are then purified and selected for using antigen markers to isolate purified T-cells. The T-cells are activated and undergo a gene transfer to express the chimeric antigen receptor (CAR) through the immunological reprogramming process. During a week-long expansion phase in parallelized small bioreactor units, the T-cells proliferate until they are comprised primarily of successfully modified T-cells. Due to the personalized nature of CAR-T cell therapy, all doses must be contained in single use reactors and facilities in order to prevent patient cross-contamination. Once T-cells have been harvested, they are processed, formulated and concentrated in a resuspension solution, after which they will be cryopreserved and transported back to the original hospital or clinic for infusion. This process design results a yearly production of 3,000 individual CAR-T doses each year.