Simultaneous analysis of cell-borne and soluble biomarkers by high throughput acoustic flow cytometry

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Project Summary Analysis of complex cellular processes in understanding disease, development of drugs, or testing efficacy of therapies often requires dynamic measurements of both cellular markers and cellular products. The ability to synchronously and rapidly conduct such measurements in complex cell- containing samples using a single instrumental analysis will be useful and beneficial across biotechnology. The goal of this project is to enable such multiplex analysis of T-cell activation using immunophenotyping and quantitative monitoring of their secreted cytokines as an important model system. High throughput measurement of samples of suspended cells is made possible using BennuBio’s Velocyt® multi-stream acoustic flow cytometry system. The primary innovation of this project is the implementation of negative acoustic contrast particles (NACPs) as a bead-based immunoassay platform that significantly extends the analytical capabilities of the Velocyt® instrument for simultaneous analysis of cell-associated and soluble biomarkers. In contrast to cells, which migrate to, and are analyzed at, discreet positions within the Velocyt® flow stream,NACPs are separated to distinct positions in the flow stream, allowing for easy discrimination between bead and cells in a single sample. In this proof-of-concept Phase I research, we aim to conduct simultaneous measurement of the T-cell activation marker CD25 in both the CD4 and CD8 population at the same time that we measure secreted interferon-γ and tumor necrosis factor-α . While this demonstration will be of significance to the development of pharmaceuticals for treatment of a range of immunological and infectious diseases that involve T-cell activation, it will also provide a firm basis by which to design and commercialize customizable NACP-based reagent kits to address a wide range of disciplines across biotechnology.