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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

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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