CAR-T cells are becoming a more common therapeutic approach for targeting of hematological disorders and solid tumors. The immunological determinants of CAR-T
efficacy need comprehensive investigation, from starting material to final product, covering different CAR constructs, transfection methods and targets.
In this study, DURAClone off-the-shelf dry antibody panels were used in our research as pre-configured assays or expanded with the addition of CAR-specific probes and liquid
antibodies. Viable CD45+ leukocyte and CD3+ T cell count, leukocyte composition (CD3/CD19/CD56/CD14/CD16) and the T cell (CD3/CD4/CD8) maturation profile (CD45RA,
CCR7, CD27, CD57) including expression of checkpoint markers PD-1, LAG-3 and TIM-3 were assessed. Data was acquired on a CytoFLEX flow cytometer and data analysis was
performed on the Cytobank platform with manual gating and advanced machine learning assisted tools. Cell count and viability was assessed in the different sample types of CAR-T cell characterization as well as the cellular composition of T/B/NK-cells, monocytes, T-cell memory and PD-1 in starting PBMC material and T-cell enriched fraction. Custom CAR-T cell panels with open channel for drop-in CAR-specific probe was used to assess BCMA-CAR expression, T cell memory maturation and checkpoint markers in Mock control CAR-T cells and BCMA CAR-T cells.