Abstract: Flow cytometry cell profiling is a laboratory technique that relies on multiparameter flow cytometry to characterize immune cell heterogeneity. Despite its common use as a diagnostic tool in monitoring immunodeficiencies, cell profiling also plays a role in ex vivo cell manufacturing, especially for stem-cell derived therapeutics. In particular, the precise identification and characterization of specific cell populations provides important information regarding the composition of the raw cellular material. In turn, this knowledge can help control the quality of the raw material to the required standards for manufacturing and greatly increase the quality and consistency of the final therapeutic. This is especially crucial in autologous therapies where the patient variability can significantly compromise quality and therapeutic efficacy. With the advent of standard of GMP guidances, drug manufacturers are striving to deliver personalized medicine quickly. Thus, the importance of rapid and high-throughput tests in evaluating patient cell type variation is paramount. A rapid, flexible cell profiling platform was developed using fluorescence-conjugated antibodies alongside gating logic and cell analysis. The entire method was designed to be run in less than 30 minutes with clearly distinguishable cell subpopulations and validated using varying sample types CAR-T, PBMCs, and more. We successfully utilized this platform to perform cell profiling of a pool of healthy donor samples, elucidating the relative abundance of CD3+ T lymphocytes, T helper cells, T cytotoxic cells, B lymphocytes and NK lymphocytes. However, this platform has the flexibility to be adapted for different cell profiling requirements and is rapid enough to be performed throughout the manufacturing process. With the availability of a cell profiling platform, this tool highlights the utility of multiparameter flow cytometry for ex vivo cell manufacturing of autologous and allogeneic cell therapies.
