US4661444A
Expired / Lapsed
US
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Homogeneous immunoassays employing double antibody conjugates comprising anti-idiotype antibody
Assignee: ORTHC DIAGNOSTIC SYSTEMS Inc A NJ CORP
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Filing: 1984-03-29
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Granted: 1987-04-28
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Expires: 2004-03-29
Abstract
Abstract Homogeneous immunoassays employing double ligand binding conjugates comprising an anti-idiotype binding partner. The anti-idiotype binding partner competes with ligand for an insolubilized ligand specific binding partner. Inhibition of binding of the conjugate to the insolubilized binding partner due to ligand binding permits the second binding partner in the conjugate to bind with insolubilized label means. Thus, detection of label is inversely related to the presence of ligand in the sample.
Inventors
- 👤 Conan K. N. Li
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 15:26
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6854; G01N33/686; G01N33/53; G01N33/543; G01N33/54306; Y; Y10; Y10S; Y10S435/00; Y10S435/965; Y10S436/00; Y10S436/80
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
55 words
An immunoassay reagent system for detecting a ligand comprising: an insolubilized ligand binding partner; a double binding partner conjugate comprising an anti-idiotype binding partner, capable of blocking said ligand from binding with said insolubilized ligand binding partner, coupled to a second binding partner; and insolubilized label means for which said second binding partner is specific.
Claim 2
Dependent Claim
55 words
An immunoassay reagent system for detecting a ligand comprising: an insolubilized ligand binding partner; a double binding partner conjugate comprising an anti-idiotype binding partner, capable of blocking said ligand from binding with said insolubilized ligand binding partner, coupled to a second binding partner; and insolubilized label means for which said second binding partner is specific.
Claim 3
Dependent Claim
57 words
The immunoassay reagent system as provided in claim 1 wherein said label means is selected from the group of labels having a characteristic spectral emission consisting of fluorescent molecules, phosphorescent molecules, and chemiluminescent molecules and wherein binding of the second binding partner to said label means results in a detectable change of said label's characteristic spectral emission.
Claim 4
Dependent Claim
38 words
The immunoassay reagent system as provided in claim 1 wherein said label means is an enzyme whose activity upon a substrate to produce a detectable product is altered upon binding of the second binding partner to said enzyme.
Claim 5
Dependent Claim
25 words
The immunoassay reagent system as provided in claim 3 wherein said enzyme is alkaline phosphatase, said substrate is p-nitrophenyl phosphate, and said product is p-nitrophenol.
Claim 6
Dependent Claim
25 words
The immunoassay reagent system as provided in claim 3 wherein said enzyme is alkaline phosphatase, said substrate is 4-methylumbelliferyl phosphate, and said product is 4-methylunbelliferone.
Claim 7
Dependent Claim
26 words
The immunoassay reagent system as provided in claim 3 wherein said enzyme is urease, said substrate is substantially ammonia free urea, and said product is ammonia.
Claim 8
Independent Claim
214 words
A method for detecting a ligand in an aqueous sample comprising the steps of: (a) contacting said sample suspected of containing the ligand to be detected with an insolubilized ligand binding partner specific for said ligand; (b) adding to said sample a double binding partner conjugate comprising a first binding partner of first specificity coupled to a second binding partner of second specificity each binding partner being immunologically reactive, and wherein said first binding partner is an anti-idiotype or anti-framework binding partner specific for said insolubilized ligand binding partner whereby the binding of either said ligand or said first binding partner to said insolubilized binding partner blocks the binding of said first binding partner or said ligand respectively; (c) contacting said sample with support means having label means attached thereto, said second binding partner being specific for said label means whereby binding of said second binding partner to said label means alters a detectable characteristic of said label means, said support means being maintained at a distance sufficiently large from said immobilized binding partner to substantially prohibit simultaneous binding of said binding partner conjugate to both said insolubilized binding partner and said label means; (d) detecting said detectable characteristic of label means and relative same to the presence of said ligand to be detected.
Claim 9
Independent Claim
214 words
A method for detecting a ligand in an aqueous sample comprising the steps of: (a) contacting said sample suspected of containing the ligand to be detected with an insolubilized ligand binding partner specific for said ligand; (b) adding to said sample a double binding partner conjugate comprising a first binding partner of first specificity coupled to a second binding partner of second specificity each binding partner being immunologically reactive, and wherein said first binding partner is an anti-idiotype or anti-framework binding partner specific for said insolubilized ligand binding partner whereby the binding of either said ligand or said first binding partner to said insolubilized binding partner blocks the binding of said first binding partner or said ligand respectively; (c) contacting said sample with support means having label means attached thereto, said second binding partner being specific for said label means whereby binding of said second binding partner to said label means alters a detectable characteristic of said label means, said support means being maintained at a distance sufficiently large from said immobilized binding partner to substantially prohibit simultaneous binding of said binding partner conjugate to both said insolubilized binding partner and said label means; (d) detecting said detectable characteristic of label means and relative same to the presence of said ligand to be detected.
Claim 10
Dependent Claim
52 words
The method as provided in claim 7 wherein said label means is selected from the group consisting of fluorescent molecules, phosphorescent molecules, chemiluminescent molecules and light scattering particles whereby immunological reaction between said second binding partner and said label means results in alteration of detectable spectral characteristics associated with said label means.
Claim 11
Dependent Claim
40 words
The method as provided in claim 7 wherein said label means is an enzyme whose capability to produce detectable product from a substrate added to the sample is altered upon immunological reaction between said enzyme and said second binding partner.
Claim 12
Dependent Claim
35 words
The method as provided in claim 9 wherein said enzyme is alkaline phosphatase, said substrate is p-nitrophenyl phosphate, and the detecting step comprises detecting increasing light extinction based on increasing levels of the product p-nitrophenyl.
Claim 13
Dependent Claim
29 words
The method as provided in claim 9 wherein said enzyme is alkaline phosphatase, said substrate is 4-methylumbelliferyl phosphate, and the detecting step comprises detecting the presence of 4-methylumbelliferone fluorimetrically.
Claim 14
Dependent Claim
35 words
The method as provided in claim 9 wherein said enzyme is urease, said substrate is substantially ammonia free urea, and the detecting step comprises detecting pH changes associated with the presence of the product ammonia.
Claim 15
Dependent Claim
41 words
In homogeneous immunoassays employing insolubilized binding partners for the detection of ligands in an aqueous sample, the improvement comprising: utilizing a double binding partner conjugate to compete with said ligand for said insolubilized binding partner and specific for insolubilized label means.
Claim 16
Dependent Claim
41 words
In homogeneous immunoassays employing insolubilized binding partners for the detection of ligands in an aqueous sample, the improvement comprising: utilizing a double binding partner conjugate to compete with said ligand for said insolubilized binding partner and specific for insolubilized label means.
References Cited (Backward Citations)
US4433059A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1990-04-27 | 1990-10-27 | 1991-04-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-04-27 | 1994-10-27 | 1995-04-28 | $3760.00 | Window Closed / Lapsed |
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