Streamlined measurement of CAR T cell concentration, size, viability, and two-color phenotyping of CAR T cells during manufacturing.

The success of CD19-targeted chimeric antigen receptor (CAR) T cell therapies has led to an exponential increase in the number of patients treated and new CAR T products being advanced. Therefore, there is a strong need to develop streamlined platforms that allow rapid, cost-effective, and accurate measurement of the key characteristics of CAR T cells during manufacturing, i.e., cell numbers, cell size (fL), viability, and basic phenotype. In this study, we compared the novel benchtop cell analyzer Moxi Go II (ORFLO Technologies), which enables simultaneous evaluation of all the above parameters, with the current standards, Multisizer Coulter Counter (cell counter) and BD LSR Fortessa (flow cytometer). Our results demonstrated that the Moxi GO II can accurately measure cell numbers and cell size (fL) while simultaneously assessing simple two-color flow cytometry parameters such as CAR T cell viability and CD4 or CAR expression. These measurements are comparable with the gold standard instruments, demonstrating that Moxi GO II is a promising platform for quickly monitoring CAR T cell growth and phenotype in research grade and clinical samples.