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T cell stimulation assays are commonly used in invitro pharmacological assays to understand immunomodulatory functions of drugs in development.^1–3 Assays read outs are common activation markers measured by flow cytometry and cytokines secretion detected in culture supernatants. In addition to monitoring activation markers, cytokines and their expression are critical for biomarker discovery and essential in understanding immune biology in addition to being commonly used as a read out in T cell stimulation.

In this work, we report CD25 CD69 expression after stimulation at 24,48,72h and IL-4, IL-2, TNFa, IFNg production from 33 healthy donors. We used a subset of samples to compare 4 technologies used in drug discovery; Meso Scale Discovery [MSD], Cytometric Bead Array [CBA] and Sartorius Q Beads Plexscreen. Through experimental data analysis, several assay features were …