For the detection and monitoring of CAR T cells in patients, two main techniques are employed: PCR and flow cytometry. Flow cytometry, however, stands out for its ability to determine the phenotype of CAR T cells. This feature of flow cytometry is vital for assessing the expansion and persistence of CAR T cells in patients, aligning with the FDA’s draft guidance recommendations. The role of flow cytometry extends beyond detection to playing a pivotal role in monitoring CAR T cells during clinical trials. It enables investigators to gain deeper insights into the behavior of CAR T cells post-infusion and their interaction with the patient’s endogenous immune system. Understanding these dynamics is crucial for evaluating the efficacy and safety of CAR T cell therapies.

In summary, flow cytometry is not just a tool for detection; it is a critical component in the advancement and monitoring of CAR T cell therapies, offering invaluable insights into the therapeutic potential and interaction of these engineered cells within the patient’s immune system.

In this poster, first presented at ESCCA 2023, discover how Cerba Research has implemented both autologous and allogenic CAR T assays.

Addressing a significant gap in the market, this collaboration aimed to overcome the limitations posed by the scarcity of Ki-67 expressing cells in standard biological samples. The synergy between Cerba Research’s expertise in clinical trial services and Slingshot Biosciences’ cutting-edge technology led to the creation of a custom-made Ki-67 cell mimic.

This solution not only met but exceeded the expectations in assay validation, showcasing a leap forward in flow cytometry techniques. This case study highlights the transformative impact of this project, marking a milestone in assay validation and flow cytometry applications in clinical research.

The ability of developing high-parameter assays allows for a deeper characterization of patients’ immune subsets with limited sample availability. Spectral flow cytometry is used to assess high-parameter immune profiling in global clinical trials. To date, there are no specific guidelines for Performance Qualification (PQ) of spectral flow cytometers. Here, we describe an in-house developed workflow for performing PQ of a Cytek® Aurora instrument in a clinical laboratory.

As flow cytometry is one of the go-to methods for fast and in-depth monitoring of immune cell populations at single cell level, clinical laboratories are observing an increase in assay complexity and number of samples to process. To scale up our operations in a qualitative and efficient manner, several aspects of the laboratory design were improved and are discussed below.

Being one of the leading technologies for cellular analysis, flow cytometry generates simultaneous high-throughput enumeration and individual cell characterization data. With the breakthrough of cellular immune therapies, such as CAR+ T, flow cytometry became a critical platform, not only for clinical laboratories, but also for drug developers and manufacturers.

Flow cytometry plays a crucial role in the production process of CAR+ T cells, where it is used for assessing transduction efficiency, purity, and characterization of the CAR+ T product before infusion. After manufacturing, when CAR+ T cells are infused into the patients, flow cytometry is used again in clinical laboratories to assess CAR+ T expansion, efficiency, and persistence and to monitor minimal residual disease (MRD).

In this e-book, Ans De Beuckelaer, Regional Head Flow Cytometry EU and Rowan Claeys, Clinical Biologist at Cerba Research, dive deeper into key elements of flow cytometry:

• The importance of flow cytometry in Car+T clinical research and its uses
• Key aspects in developing a CAR+ T trial
• The added value of flow cytometryin multiple myeloma, Minimal Residual Disease (MRD) assessments and multiple Myeloma phenotype
• Car+T-cell Therapy: patient treatment, targeted antigens and solutions for MRD assessments and multiple myeloma phenotype assessment.