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        Use of Primary Human Cells in High-Throughput Screens

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        Traditionally, the objective of high-throughput screening (HTS) has been to identify compounds that interact with a defined target protein as the starting point for a chemistry lead optimisation programme. To enable this it has become commonplace to express the drug target in a recombinant expression system and use this reagent as the source of the biological material to support the HTS campaign. In this chapter we describe an alternative HTS methodology with the objective of identifying compounds that mediate a change in a defined physiological end point as a consequence of compound activity in human primary cells. Rather than screening at a defined molecular target, such “phenotypic” screens permit the identification of compounds that act at any target protein within the cell to regulate the end point under study. As an example of such a screen we will describe an HTS campaign to identify compounds that promote the production of the cytokine interferon-α from human blood peripheral mononuclear cells (PBMCs) isolated from whole blood. We describe the procedures required to obtain and purify human PBMCs and the electrochemiluminescence-based assay technology used to detect interferon-α and highlight the challenges associated with this screening paradigm.
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