AN AUTOMATED 24-HOUR CAR T MANUFACTURING PROCESS

Background & Aim Upon treatment with chimeric antigen receptor (CAR) T cells, approximately 30–40% of patients with B cell malignancies show long-term remission. While this is a major advancement in treating these otherwise fatal cancers, there are drawbacks to CAR T cell therapy manufacturing that hinder further advancement of CAR T cell therapy use. Current CAR T cell manufacturing protocols require 7-14 days. During this time, a malignancy could progress rapidly and lead to death. Furthermore, studies show that prolonged ex vivo culture and expansion of CAR T cells is associated with a reduced lifespan and potency of those CAR T cells after infusion into patients. To beat rapid disease progression, increase time in remission, reduce vein-to-vein time and ultimately reduce the incidence of death due to B cell malignancies, it is important to develop protocols that enable quicker CAR T cell manufacturing while ensuring that CAR T cells can effectively kill cancerous CD19 B cells. This will also minimize the cost of CAR T cell manufacturing and make it more accessible. Methods, Results & Conclusion In this study, we describe an expedited 24-hour CAR T cell manufacturing process that uses Thermo Fisher Scientific CTS instruments, and which leverages lentiviral transduction of the CD19-CAR gene into T cells isolated from frozen leukpaks. This expedited process led to a final CAR T cell product with high purity, process recovery, as well as increased effectiveness. Upon treatment with chimeric antigen receptor (CAR) T cells, approximately 30–40% of patients with B cell malignancies show long-term remission. While this is a major advancement in treating these otherwise fatal cancers, there are drawbacks to CAR T cell therapy manufacturing that hinder further advancement of CAR T cell therapy use. Current CAR T cell manufacturing protocols require 7-14 days. During this time, a malignancy could progress rapidly and lead to death. Furthermore, studies show that prolonged ex vivo culture and expansion of CAR T cells is associated with a reduced lifespan and potency of those CAR T cells after infusion into patients. To beat rapid disease progression, increase time in remission, reduce vein-to-vein time and ultimately reduce the incidence of death due to B cell malignancies, it is important to develop protocols that enable quicker CAR T cell manufacturing while ensuring that CAR T cells can effectively kill cancerous CD19 B cells. This will also minimize the cost of CAR T cell manufacturing and make it more accessible. In this study, we describe an expedited 24-hour CAR T cell manufacturing process that uses Thermo Fisher Scientific CTS instruments, and which leverages lentiviral transduction of the CD19-CAR gene into T cells isolated from frozen leukpaks. This expedited process led to a final CAR T cell product with high purity, process recovery, as well as increased effectiveness.