US4859609A
Expired / Lapsed
US
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Novel receptors for efficient determination of ligands and their antagonists or agonists
Assignee: Genentech Inc
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Filing: 1986-04-30
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Granted: 1989-08-22
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Expires: 2006-04-30
Abstract
Abstract Hybrid receptors, produced by recombinant DNA technology, comprise (a) the ligand binding domain of a predetermined receptor and (b) a heterologous reporter polypeptide. The hybrid receptors are useful for convenient and large scale assay of biologically active ligands or their antagonists or agonists. (a) may be the extracellular domain of the receptor, or a cytoplasmic domain of a receptor or oncogene. (b) may be an enzyme. A transmembrane domain may be interposed between (a) and (b).
Inventors
- 👤 Thomas J. Dull
- 👤 Heimo Riedel
- 👤 Axel Ullrich
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 10:45
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/705; C07K14/72; C07K14/71; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/566; G01N33/68; C07K2319/00; C07K2319/01; C07K2319/033; C07K2319/32; C07K2319/90; G01N2500/00; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S530/00; Y10S530/806; Y10S530/808
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
61 words
A hybrid receptor for a ligand comprising (a) a ligand binding domain of a predetermined receptor fused at its C-terminus to the N-terminus of (b) a reporter polypeptide which is (1) heterologous to the receptor from which is obtained the ligand binding domain and which (2) undergoes a conformational change upon the binding of said ligand to the ligand binding domain.
Claim 2
Dependent Claim
61 words
A hybrid receptor for a ligand comprising (a) a ligand binding domain of a predetermined receptor fused at its C-terminus to the N-terminus of (b) a reporter polypeptide which is (1) heterologous to the receptor from which is obtained the ligand binding domain and which (2) undergoes a conformational change upon the binding of said ligand to the ligand binding domain.
Claim 3
Dependent Claim
21 words
The hybrid receptor of claim 1 wherein the ligand binding domain is comprised by the extracellular domain of the predetermined receptor.
Claim 4
Dependent Claim
21 words
The hybrid receptor of claim 1 wherein the reporter polypeptide is a cytoplasmic domain of a receptor of an oncogene product.
Claim 5
Dependent Claim
13 words
The hybrid receptor of claim 1 wherein the reporter polypeptide is an enzyme.
Claim 6
Dependent Claim
12 words
The hybrid receptor of claim 4 wherein the enzyme is a phosphorylkinase.
Claim 7
Dependent Claim
21 words
The hybrid receptor of claim 1 having a transmembrane domain interposed between the ligand binding domain and the heterologous reporter polypeptide.
Claim 8
Dependent Claim
26 words
The hybrid receptor of claim 1 wherein the ligand binding domain of the predetermined receptor consists essentially of the ligand binding domain of the predetermined receptor.
Claim 9
Independent Claim
83 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a conformational change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 10
Independent Claim
83 words
A method for assaying a biologically active ligand or an antagonist or agonist for said ligand, comprising (a) providing a hybrid receptor which comprises (1) a binding domain for the ligand, antagonist or agonist and (2) a heterologous reporter polypeptide; (b) incubating the receptor with a test sample suspected to contain the ligand, antagonist or agonist; (c) detecting a conformational change in the reporter polypeptide; and (d) correlating said change with the presence of the ligand, antagonist or agonist in the test sample.
Claim 11
Dependent Claim
25 words
The method of claim 8 wherein the ligand is a polypeptide and the ligand binding domain is not the antigen binding site of an immunoglobulin.
Claim 12
Dependent Claim
22 words
The method of claim 8 wherein the change in the reporter polypeptide is a modification of the enzymatic activity of the polypeptide.
Claim 13
Dependent Claim
18 words
The method of claim 10 wherein the change in the reporter polypeptide is autophosphorylation of the reporter polypeptide.
Claim 14
Dependent Claim
33 words
The method of claim 8 for assaying a test sample suspected of containing an antagonist wherein the receptor is incubated with the test sample and a predetermined amount of ligand or ligand agonist.
Claim 15
Dependent Claim
16 words
The method of claim 8 wherein the change in the reporter polypeptide is an immunoligical change.
Claim 16
Dependent Claim
56 words
The method of claim 8 wherein the reporter polypeptide further comprises a stable free radical, fluroescent or chemiluminescent group and the change in the reporter polypeptide is detected by measuring a change in the rotational moment of the stable free radical or a change in the intensity, wavelength or polarization of the fluorescent or chemiluminescent group.
Claim 17
Dependent Claim
17 words
The method of claim 8 wherein the reporter polypeptide is capable of binding to a G protein.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-08-21 | 1993-02-20 | 1993-08-22 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-08-21 | 1997-02-20 | 1997-08-22 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-08-21 | 2001-02-20 | 2001-08-22 | $7700.00 | Window Closed / Lapsed |