A 38-color spectral panel for cost efficient characterization of T cell therapy products

Investigators must frequently apportion precious sample volumes between multiple assays to measure the viability, potency, and compositional phenotype of cell therapy products and clinical research samples. However, the use of disparate assay modalities may miss critical information which links the expression profile of cells with functional activity, especially when these activated cell populations are rare. To address this shortcoming, we describe in this short report the creation of a 38-color spectral flow cytometry panel for the phenotypic and functional characterization of antigen reactive T cell populations within cellular therapy products and clinical research samples. Our investigations identified that up to 29 potentially co-expressing markers were able to be identified on mixed sample populations using this panel, including phenotype, activation, and exhaustion markers, as well as intracellular cytokine production. Our calculations also highlight spectral flow cytometry is a more efficient use of technician time, reagent cost, and product sample usage as compared with an equivalent set of traditional flow cytometry panels.