Real-Time Potency Assay for CAR T Cell Killing of Adherent Cancer Cells

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Summary

To rigorously compare different CAR constructs and identify unwanted behavior such as tonic (antigen-independent) signaling, CAR function should be evaluated within the complex milieu of the immune synapse, using cancer cell killing as the readout.
Typical release assays (51Cr, LDH) have historically been the gold standard, but suffer from the fact that they require significant “hands-on” time and only provide end-point data.
By combining the strengths of real-time impedance monitoring (simplicity, analytical sensitivity, and objectivity) with that of live cell imaging (specificity of the readout), eSight increases the information richness of the CAR T cell‑killing assay without increasing the workload

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Quantifying CAR-T Cell Killing Efficacy by Real Time Impedance

Figure 5. (Edited) Time courses for HEK-293-CD19 killing by CD19 CAR T cells as measured by impedance.

Monitoring CAR-T Cell Killing Activity by Live Cell Imaging

Figure 3B. Killing of RFP-expressing HEK-293-CD19 cells by CD19 CAR T cells. (A) Comparison of different E:T ratios 48 hours post CAR T-cell addition. The white squares in the upper panels denote the regions that are blown up in the lower panels. The unlabeled CAR T cells are grey. (B) Comparison of different time points for a constant E:T ratio of 4:1.

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