C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 5/10 (2006.01) G01N 33/566 (2006.01) G01N 33/68 (2006.01)
Patent
CA 2255548
This invention provides a system for screening chemical compounds to identify ligands for receptors including G-protein coupled receptors. The invention exploits cells in which the receptor is coupled through a second messenger system to an ion channel that is gated by cyclic nucleotide. Receptor stimulation causes the second messenger system to produce cyclic nucleotide, which results in ion influx through the channel. By measuring ion influx fluorescently, the invention provides a rapid and convenient means for identifying receptor ligands. By providing mixed cell cultures that include cells expressing different receptor types, and by loading into those cells different fluorescent reporters of ion influx, the invention further provides a multiplexed system that accelerates the ligand identification process. Cells useful in the process, and methods for exploiting them, are described. Also described is a "tandem" screening protocol in which cells of the invention are first incubated with a ligand candidate to generate a first signal, and then with a reference agonist for the receptor target expressed by the cell. The effect of the reference agonist on the first signal can then be used to characterize candidate ligands as target receptor agonists or modulators. 42
Ballyk Barbara Ann
Catalano Concettina
Demchyshyn Lidia
Lee David K. H.
Zastawny Roman
Allelix Biopharmaceuticals Inc.
Mccarthy Tetrault Llp
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