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Development of Radioligand Binding Assays for the Motilin Receptor Using ScreenReady Targets.

Vronique Brechler, David Handfield, Martin Boissonneault, Esther Tremblay, Benoit Houle and Luc Mnard.

Introduction

Recent advances in combinatorial chemistry and genomics programs have led to an unprecedented increase in both the number of compounds and potential drug targets that can be screened. The development of automated liquid handling and microplate assay systems has helped to meet the demand for increased throughput. Concomitantly, the introduction of homogeneous, non-separation, assay systems has eased the development of fully automated assays, especially for G protein-coupled receptors (GPCRs). GPCRs are composed of a large family of membrane-bound receptors that are characterized by the presence of seven hydrophobic membrane-spanning regions (Watson et al ., 1994). These receptors are involved in numerous aspects of cell signaling and regulation, and represent one of the major classes of the current therapeutic drug targets for the drugs sold today. Traditionally, pharmacological binding assays for these receptors have been performed using filtration assays that are difficult to automate.

For over ten years, PerkinElmer Life Sciences has been actively involved in the development of expression systems and novel assays for GPCRs. Given the current market need for simple homogeneous assays for GPCRs, PerkinElmer developed a novel line of reagents that are ready-to-use and pre-formatted for detection of ligand-receptor interactions. Using FlashPlate microplates as the homogeneous platform, we have developed a proprietary coating procedure to allow the stable and efficient immobilization of recombinant GPCRs on FlashPlate microplates. Assays are microformatted to a minimum volume of 25 μl, thereby reducing the costs of reagents and radioactive waste management, but can easily be formatted to 50 μl.

Membrane-coated FlashPlate microplates are suppli
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