| abstract |
A drug discovery method for selecting a compound selected from the group consisting of a small organic substance, a biopharmaceutical and an antibody or part thereof. The method comprises the steps of i) expressing one or more receptors on a cell membrane, such as, e.g., an exterior cell surface of a cell, ii) contacting one or more expressed receptors with a test compound or a selection of test compounds (libraries) and iii) selecting one or more compounds based on its ability to bind one or more receptors. The step of expressing the one or more receptors comprises capturing one or more receptors on the exterior cell surface in a conformation that predominantly enables binding or interaction with a ligand, and the conformation that predominantly enables binding or interaction with a ligand is provided by modification of one or more receptors by a method comprising at least one of the following: (a) fusion with any protein which keeps the receptor in the desired conformation such as, e.g. an arrestin, a modified arrestin, a G-protein or a modified G-protein, (b) site-directed mutagenesis, and (c) deletion. The receptors may be captured on the exterior cell surface by at least one of the following: (d) interaction of the receptor with a scaffolding protein, optionally, with a scaffolding protein network and (e) means for blocking receptor internalization, e.g. by co-expression of a mutated dynamin or a modified arrestin or by use of chemicals such as, e.g., sucrose and/or Tris. |