Millipore Technical Publications |
Detection of Allosteric Potentiators for GPCRs with Millipore’s GPCRProfiler Platform | ||
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| An increasing number of allosteric modulators have been described for GPCRs, and the potential is high for more allosteric modulators of GPCRs to develop into viable therapeutic compounds. However, specialized techniques are required to detect allosteric modulators in an HTS setting. In particular, the detection of allosteric potentiators requires a robust readout capable of yielding reproducible signals at low concentrations of agonist (EC20 maximum).We have characterized methods for detection and pharmacological characterization of allosteric potentiators with our ChemiScreen™ system, in which recombinant GPCRs (currently over 130 available) are expressed in proprietary host cell lines that express endogenous promiscuous Gα15/16 to yield a common calcium readout regardless of the coupling status of the GPCR.We have characterized allosteric modulators for 2 targets, GABAB, and A1, expressed in ChemiScreen™ cell lines by calcium flux assay, and found comparable pharmacology to previous reports. In addition, we designed a three-addition protocol to screen compounds for allosteric modulators in addition to agonists and antagonists, in which the first addition is the compound, the second addition is an EC20 amount of standard agonist, and the third addition is an EC80 amount of standard agonist.We tested a small library consisting of known compounds against GABAB and A1, and were able to detect the correct allosteric modulator for each. Therefore, the ChemiScreen™ system employed by the GPCRProfiler service provides a platform for detection of allosteric potentiators for a broad panel of GPCRs with a common calcium readout. Click PDF above to view entire poster. |