OR13

CAR-T cell therapies for cancer have seen significant clinical advances, but analyzing the potency and identity of these drug products remains challenging. As new CAR-T products and production methodologies are developed, rigorous evaluation of each batch's relative potencies is increasingly important. Two MOA-based bioluminescent assays have been developed to overcome challenges in quantitative assessment of T cell therapies. Both assays rely on split NanoBiT® luciferase technology.

The HiBiT Target Cell Killing Assay involves incubating CAR-T cells with target cells stably expressing HiBiT. This results in target cell lysis and release of HiBiT proteins, which bind with high affinity to LgBiT in the detection reagent, forming functional NanoBiT® luciferase and producing light in proportion to target cell lysis.

Lumit® Immunoassays are a simple and homogeneous method for measuring cytokine production from engineered T cells. Monoclonal antibodies are covalently labeled with LgBiT and a low-affinity tag, SmBiT. When the antibodies are brought together on a common analyte, such as a cytokine, SmBiT and LgBiT can complement to form NanoBiT® luciferase and produce light in proportion to the concentration of analyte in the sample. Lumit® Immunoassays can be run directly on cells, removing media transfer, dilution, and wash steps that can be common sources of error.

When autologous CAR-T products are being produced in parallel, it is essential to verify the identity of each product and ensure that it is administered to the correct patient. An easy workflow for STR analysis can be used to document the chain of identity for CAR-T products.