	US 7,541,151 B2
	Single-cell biosensor for the measurement of GPCR ligands in a test sample
Lawrence S. Barak, Durham, N.C. (US); Michael A. Shetzline, Chapel Hill, N.C. (US); Robert H. Oakley, Durham, N.C. (US); and Marc G. Caron, Hillsborough, N.C. (US)
Assigned to Duke University, Durham, N.C. (US)
Filed on Jun. 04, 2002, as Appl. No. 10/161,916.
Application 10/161916 is a continuation in part of application No. 09/631468, filed on Aug. 03, 2000, abandoned.
Application 09/631468 is a continuation of application No. 09/233530, filed on Jan. 20, 1999, granted, now 6,110,693.
Application 09/233530 is a continuation of application No. 08/869568, filed on Jun. 05, 1997, granted, now 5,891,646.
Claims priority of provisional application 60/295945, filed on Jun. 05, 2001.
Prior Publication US 2009/0075314 A1, Mar. 19, 2009
This patent is subject to a terminal disclaimer.
Int. Cl. G01N 33/566 (2006.01); G01N 21/76 (2006.01); G01N 33/53 (2006.01)

U.S. Cl. 435—7.1 [435/7.2; 435/7.21; 436/172; 436/501; 530/350; 536/23.4]

35 Claims

1. A method of detecting the presence or absence of a G protein-coupled receptor (GPCR) ligand in a test sample, comprising the steps of:

(a) providing a cell comprising a GPCR and a detectably labeled arrestin, wherein the cellular distribution of the detectably labeled arrestin changes in response to activation of the GPCR;

(b) determining the cellular distribution of the detectably labeled arrestin in the absence of a test sample;

(d) determining the cellular distribution of the detectably labeled arrestin in the presence of the test sample; and

(e) comparing the cellular distribution of the detectably labeled arrestin in the presence of the test sample to the cellular distribution of the detectably labeled arrestin in the absence of the test sample,

wherein a change in the cellular distribution of the detectably labeled arrestin in the presence of the test sample in response to a GPCR ligand as compared to the cellular distribution of the detectably labeled arrestin in the absence of the test sample indicates the presence of a GPCR ligand in the test sample.