Discussion

The detection and quantification of secreted proteins provide key information about signaling pathways, and specifically the dysfunctional signaling implicated in many disease areas such as cancer, immune disorders and aging. These signaling pathways are important therapeutic targets across the drug discovery processes from primary screening to toxicity profiling. In an immunological example, secretion of cytokines from one cell type will trigger a cascade of downstream events in other cell types. The ability to simultaneously measure not only multiple cytokine secretion, but also cell proliferation and cell phenotye can maximize the contextual and correlative value of screeningdata. The iQue® Screener is the only screening platform capable of simultaneous profiling of cytokine secretion and cell proliferation from a single sample by measuring beads and cells mixture in a single assay. The iQue Screener platform features the unique ability to distinguish cells and beads in a high throughput flow format with zero dead volume, enabling the ability to screen compound libraries in multiplexed fashion. In this case study, PBMCs were used as the biological model with phytohaemagglutinin (PHA) to stimulate cytokine secretion and T cell proliferation. IntelliCyt’s MultiCyt® QBeads™ and Cell Proliferation Dye Panel were used to simultaneously detect cytokines and cell proliferation, respectively, in addition to the immunophenotyping (CD3 and CD8) in the same multiplexed assay. The optimized beads and cells combination workflow is uniquely suited to the iQue Screener, and may promote the adoption of primary cells in profiling/screening in the drug discovery industry.