{"search_session":{},"preferences":{"l":"en","queryLanguage":"en"},"patentId":"022-374-860-007-988","frontPageModel":{"patentViewModel":{"ref":{"entityRefId":"022-374-860-007-988","entityRefType":"PATENT"},"entityMetadata":{"linkedIds":{"empty":true},"tags":[],"collections":[{"id":10707,"type":"PATENT","title":"Duke Univ Patent Portfolio","description":"","access":"OPEN_ACCESS","displayAvatar":true,"attested":false,"itemCount":9941,"tags":[],"user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"notes":[{"id":8224,"type":"COLLECTION","user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"text":"
Search Applicants and Owners separately :' Duke Univ\"; \" Univ Duke\"; \" Uni* Duk*\"; \" Duk* Uni*\"; ' Duke University\" ; \" University Duke\".
Select more for logical variants. Add to collection. Select all patents in the collection and expand by simple families. Add to collection. Total patents: 8489
Search Applicants and Owners separately :' Duke Univ\"; \" Univ Duke\"; \" Uni* Duk*\"; \" Duk* Uni*\"; ' Duke University\" ; \" University Duke\".
Select more for logical variants. Add to collection. Select all patents in the collection and expand by simple families. Add to collection. Total patents: 8489
(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;\n
(b) determining the cellular distribution of the detectably labeled arrestin in the absence of a test sample;\n
(c) exposing the cell to the test sample;\n
(d) determining the cellular distribution of the detectably labeled arrestin in the presence of the test sample; and\n
(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,\n
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."],"number":1,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution of the detectably labeled arrestin is determined at different time points after exposure to the test sample."],"number":2,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution of the detectably labeled arrestin is determined after exposure to different concentrations of the test sample."],"number":3,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution of the detectably labeled arrestin is quantified."],"number":4,"annotation":false,"claim":true,"title":false},{"lines":["The method claim 4, wherein a concentration of the GPCR ligand in the test sample is quantified by comparing localization of the detectably labeled arrestin in plasma membrane, clathrin-coated pits, endocytic vesicles, or endosomes as compared to cytosol for the detectably labeled arrestin in the presence of the test sample to localization of the detectably labeled arrestin in plasma membrane, clathrin-coated pits, endocytic vesicles, or endosomes as compared to cytosol for the detectably labeled arrestin in the presence of a known concentration of the ligand."],"number":5,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 5, wherein when a change in the concentration of the GPCR ligand in the test sample is observed over a period of time as a result of a compound that alters the concentration of the ligand indicates the presence of a compound in the test sample that alters the concentration of the ligand."],"number":6,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample is or is derived from serum, tissue, blood, or urine."],"number":7,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1 wherein the GPCR is CCK-B (SEQ ID NO: 13) or CCK-A (SEQ ID NO: 11)."],"number":8,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 8, wherein the test sample was derived from a patient with hypergastrinemia."],"number":9,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the ligand is gastrin, preprogastrin, cleaved preprogastrin, gastrin-34, gastrin-17, pentagastrin, progastrin, glycine-extended gastrin-17, glycine-extended gastrin-34, gastrin-71, gastrin-6, hG17, a compound with an amidated tetrapeptide of the sequence Trp-Met-Asp-Phe-NH2 (SEQ ID NO: 15), or an isoform of gastrin capable of binding to a gastrin receptor."],"number":10,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 10, wherein the gastrin concentration is less than 10 nM."],"number":11,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the GPCR is a muscarinic receptor."],"number":12,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the ligand is acetylcholine."],"number":13,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the labeled arrestin is localized in the cytosol, plasma membrane, clathrin-coated pits, endocytic vesicles, or endosomes."],"number":14,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein an increase in a local concentration of the detectably labeled arrestin in plasma membrane, clathrin-coated pits, endocytic vesicles, or endosomes as compared to cytosol results in an increase in a local signal intensity."],"number":15,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 15, wherein the local signals intensity is increased in the presence of increased concentration of ligand in the test sample."],"number":16,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein a signal intensity of the labeled arrestin in the plasma membrane, clathrin-coated pits, endocytic vesicles, or endosomes is increased as compared to a level of signal intensity in the cytosol."],"number":17,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the detectable label is a radioisotope, an epitope tag, an affinity label, an enzyme, a fluorescent group, or a chemiluminescent group."],"number":18,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution is visualized by flow cytometry."],"number":19,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution is visualized by fluorescence confocal microscopy."],"number":20,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein a computer analyzes an image of the cellular distribution."],"number":21,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the distribution of the detectably labeled arrestin is quantified."],"number":22,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample comprises a ligand of the GPCR."],"number":23,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample comprises an antagonist of the GPCR."],"number":24,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample further comprises acetylcholine and acetylcholinesterase."],"number":25,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample further comprises an agonist of the GPCR."],"number":26,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the ligand has been identified."],"number":27,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the test sample is heterogeneous."],"number":28,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution is determined after 15-30 minutes of exposure to the test sample."],"number":29,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cellular distribution is determined after 1 hour of exposure to the test sample."],"number":30,"annotation":false,"claim":true,"title":false},{"lines":["The method of claim 1, wherein the cell is exposed to the test sample at a temperature of 37° C."],"number":31,"annotation":false,"claim":true,"title":false},{"lines":["A method of detecting the presence or absence of a G protein-coupled receptor (GPCR) ligand in a test sample, comprising the steps of:\n
(a) providing a single cell biosensor comprising a cell which overexpresses a detectably labeled arrestin and at least one GPCR, wherein the cellular distribution of the detectably labeled arrestin changes in response to activation of the GPCR;\n
(b) determining the cellular distribution of the detectably labeled arrestin in the absence of a test sample;\n
(c) exposing the biosensor to the test sample;\n
(d) determining the cellular distribution of the detectably labeled arrestin in the presence of the test sample; and\n
(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,\n
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."],"number":32,"annotation":false,"claim":true,"title":false},{"lines":["A method of continuous screening for the presence or absence of G protein-coupled receptor (GPCR) ligands in a test sample, comprising the steps of:\n
(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;\n
(b) determining the cellular distribution of the detectably labeled arrestin in the absence of a test sample;\n
(c) exposing the cell to the test sample;\n
(d) determining the cellular distribution of the detectably labeled arrestin in the presence of the test sample;\n
(e) comparing the cellular distribution of the detectably labeled arrestin in the presence of the test sample in response to a GPCR ligand 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 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; and\n
(f) replacing the cell with another cell comprising a GPCR and a detectably labeled arrestin and repeating steps (b)-(e) thereby continuously screening for the presence or absence of G protein-coupled receptor (GPCR) ligands in a test sample."],"number":33,"annotation":false,"claim":true,"title":false},{"lines":["A method of detecting the presence or absence of a compound that modulates G protein-coupled receptor (GPCR) internalization in a test sample, comprising the steps of:\n
(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 and wherein when the GPCR is activated and internalized the GPCR and the detectably labeled arrestin form a detectably labeled arrestin/GPCR complex;\n
(b) determining the cellular distribution of the detectably labeled arrestin in the absence of a test sample;\n
(c) exposing the cell to an agonist of the GPCR,\n
(d) exposing the cell to the test sample;\n
(e) determining the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample; and\n
(f) comparing the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample to the cellular distribution of the detectably labeled arrestin in the absence of the test sample,\n
wherein a change in the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample as compared to the cellular distribution of the detectably labeled arrestin/GPCR complex in the absence of the test sample indicates the presence of a compound that modulates GPCR internalization in the test sample."],"number":34,"annotation":false,"claim":true,"title":false},{"lines":["A method of detecting the presence or absence of a compound that modulates G protein-coupled receptor (GPCR) internalization in a test sample, comprising the steps of:\n
(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 and wherein when the GPCR is activated and internalized the GPCR and the detectably labeled arrestin form a detectably labeled arrestin/GPCR complex;\n
(b) determining the cellular distribution of the detectably labeled arrestin in the presence of an agonist of the GPCR and the absence of a test sample;\n
(c) exposing the cell to the test sample;\n
(d) determining the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample; and\n
(e) comparing the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample to the cellular distribution of the detectably labeled arrestin in the absence of the test sample,\n
wherein a change in the cellular distribution of the detectably labeled arrestin/GPCR complex in the presence of the test sample as compared to the cellular distribution of the detectably labeled arrestin/GPCR complex in the absence of the test sample indicates the presence of a compound that modulates GPCR internalization in the test sample."],"number":35,"annotation":false,"claim":true,"title":false}]}},"filters":{"npl":[],"notNpl":[],"applicant":[],"notApplicant":[],"inventor":[],"notInventor":[],"owner":[],"notOwner":[],"tags":[],"dates":[],"types":[],"notTypes":[],"j":[],"notJ":[],"fj":[],"notFj":[],"classIpcr":[],"notClassIpcr":[],"classNat":[],"notClassNat":[],"classCpc":[],"notClassCpc":[],"so":[],"notSo":[],"sat":[]},"sequenceFilters":{"s":"SEQIDNO","d":"ASCENDING","p":0,"n":10,"sp":[],"si":[],"len":[],"t":[],"loc":[]}}