P-150-T - New Methods for MOA-based Potency Testing and CAR-T Cell Characterization

Potency testing of cell therapy products requires mechanism-of-action (MOA)-based assays that accurately reflect biological activity. Current methods often lack sensitivity, scalability, or flexibility across diverse cell backgrounds. We developed a suite of assays leveraging HiBiT bioluminescent technology to quantify target cell killing (TCK) by CAR-T and other effector cells. HiBiT-expressing target cells release detectable signal upon lysis, enabling sensitive, gain-of-signal TCK measurement across stable and transiently transfected cell lines cells via ViaScript mRNA transfection. CRISPR-generated CD19/CD20 knockout panels validated tandem- and dual-CAR T cell specificity. Complementary Lumit® immunoassays quantified CAR-T-secreted cytokines without washing steps, and metabolic profiling assays tracked T cell activation states. Together, these tools provide a comprehensive, MOA-aligned platform for potency testing and characterization of cell therapy products.