Patent ID: 8551718

Claim:
An assay method for screening for antagonists and agonists of 5-HT 2A receptor, histamine H1 receptor or adrenergic alpha 1b receptor by a reporter gene based assay in a microtitre plate comprising: a) transfecting chinese hamster ovary (CHO) cells with a specific G-Protein Coupled Receptor (GPCR) coupled to a G protein containing Gαq subunit construct, wherein the specific GPCR is selected from 5-HT 2A receptor, Histamine H1 receptor and adrenergic alpha 1b receptor along with excess of cAMP response element luciferase (CRE-luc) reporter gene with 1-Propanaminium, N-(3-[(4[(3-aminopropyl) amino) butyl) amino) propyl) amino)-3-oxopropyl)-N,N-dimethyl-2,3-bis(((9z)-1-oxo-9-octadecenyl)oxy)-, 2,2,2-trifluoroacetate (1:1), mixt. with 1-(((2-aminoethoxy)hydroxyphosphinyl)oxy)methyl)-1,2-ethanediyl di-(9z)-9-octadecenoate and culturing the resultant recombinant CHO cells in a suitable medium containing a selection of suitable antibiotics; b) selecting individual colonies of recombinant CHO cells exhibiting maximum luciferase activity to both forskolin and an agonist of said specific GPCR and analyzing the expression of said specific GPCR; c) plating and culturing the selected recombinant CHO cells in a microtitre plate; d) incubating said cultured, recombinant CHO cells with increasing concentrations of a test compound at 37° C. in CO 2 incubator and evaluating the effect of the test compound as an antagonist or agonist of said specific GPCR; e) after incubation of the cells is over, removing the medium, washing the cells with buffer and lysing the cells with lysis buffer; and f) measuring the CRE-Luc reporter gene activity in individual wells of the microtitre plate.