White Paper:  Phenotypic and Functional Characterization of CAR-T Cells with Advanced Flow Cytometry and Live-Cell Analysis

One key area in cancer immunotherapeutic advances, is the use of gene-modified cell therapies, with chimeric antigen receptor (CAR) T cells leading the field. The CAR construct is designed to interact with a specific surface epitope or antigen present on the tumor cell, which when in close proximity, enables the T cell to kill the tumor cell. Where specific antigens can be identified on the tumor cells, CAR-T cells display targeted effects and, as they are sourced from the patient (known as autologous therapy), there is a lack of rejection.

Despite this progress, there are some obstacles, such as the high cost and technical difficulties of phenotyping, profiling and purifying immune cells. Also, as some patients are not highly responsive to treatment, uncovering the mechanistic basis for differing outcomes is an active area of research.

Research is progressing to explore the introduction of CAR constructs into alternative immune cells, such as CAR-NK or CAR-macrophages, and to investigate gene modified cells which target solid tumors.

There is a focus on improving construct longevity, selectivity, manufacturing and delivery to the patient, to enable efficiencies of clinical cell production and a potential switch to the use of allogeneic, off-the-shelf products.


This white paper introduces:

  • The process of manufacturing and expanding cell therapy products, with a focus on CAR-T cells.
  • Key iQue® Advanced High Throughput Flow Cytometry Platform and Incucyte® Live-Cell Analysis System, in vitro assays used to phenotype and assess function of CAR-T cells.
  • Characterization of T cell therapies.


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