(1025-B) Automating selection of engineered target cell lines for Chimeric Antigen Receptor T cells
Monday, February 5, 2024
2:00 PM – 3:00 PM EST
Location: Exhibit Halls AB
Abstract: A fundamental component for generating CAR (Chimeric Antigen Receptor) T cell therapies is the development of high-quality reagents, including selecting appropriate target cell lines for both in vitro and in vivo assays. Here, we developed an assay for QC (Quality Control) of engineered target cell lines using automated liquid handlers. We tested this platform in the assessment of a series of engineered variants of the AML (Acute Myeloid Leukemia) cell line MV-4-11. A sequential series of engineering steps were performed, starting from the parental, un-engineered cells. In the first iteration, we added a luciferase reporter gene using lentiviral transduction. In the second step, both MHC class I and II were knocked out. In the final step we knocked out the antigen of interest. Each step resulted in multiple clones that needed to be assessed to identify which clones to carry forward to the next step. We designed assays to determine how CAR-T cells respond to the parental and engineered cells, including parameters such as CAR-T cell proliferation, cytokine secretion, and cytotoxicity. To measure T cell proliferation and cytokine secretion we used the human T cell Activation cell and cytokine Kit from SartoriusTM and a luciferase cytotoxicity assay to detect direct killing of target cell lines. Lastly, we also performed an IncucyteTM assay to measure the growth kinetics of the target cell lines. To set up and perform the four assays we used a combination of three automated liquid handlers; the Agilent Bravo, the Integra Assist Plus, and the Formulatrix Mantis. This new process allowed us to select the appropriate MV-4-11 target cell line more rapidly from the multiple clones, accelerating our ability to develop novel assays.