Patent US4486530A successfully refreshed from Google Patents.
US4486530A
Expired / Lapsed
US
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Immunometric assays using monoclonal antibodies
Assignee: Hybritech Inc
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Filing: 1981-06-24
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Granted: 1984-12-04
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Expires: 2001-06-24
Abstract
Abstract "Two-site" or "sandwich" immunometric assay techniques for determination of the presence and/or concentration of antigenic substances in fluids using monoclonal antibodies are described and compared to conventional assays using polyclonal antibodies. Also described are inhibition assays using complexes of antigens with a monoclonal antibody.
Inventors
- 👤 Gary S. David
- 👤 Howard E. Greene
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 13:37
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/577; G01N33/536; G01N33/542; G01N33/543; G01N33/54313; G01N33/576; Y; Y10; Y10S; Y10S436/00; Y10S436/80
Claims Analysis
Total Claims: 34
Claim 1
Independent Claim
90 words
In an immunometric assay process to determine the presence or concentration of an antigenic substance in a fluid sample comprising forming a ternary complex of the antigenic substance, a first antibody and a second antibody bound to the antigen at a difference site than the first antibody by contacting the sample with said first and second antibodies, the improvement comprising employing a measured amount of a monoclonal antibody having an affinity for the antigenic substance of at least about 10 8 liters/mole for each of said first and second antibodies.
Claim 2
Dependent Claim
90 words
In an immunometric assay process to determine the presence or concentration of an antigenic substance in a fluid sample comprising forming a ternary complex of the antigenic substance, a first antibody and a second antibody bound to the antigen at a difference site than the first antibody by contacting the sample with said first and second antibodies, the improvement comprising employing a measured amount of a monoclonal antibody having an affinity for the antigenic substance of at least about 10 8 liters/mole for each of said first and second antibodies.
Claim 3
Dependent Claim
66 words
A process according to claim 1 wherein a fluorescent chromophore is bonded to said first antibody and a chromophore capable of absorbing light at the wavelength emitted by the fluorescent chromophore is bonded to said second antibody, the monoclonal antibodies being capable of binding to the antigenic substance at sites which permit the light emitted by the fluorescing chromophore to be absorbed by the quenching chromophore.
Claim 4
Dependent Claim
52 words
A process according to claim 2 wherein the sample is contacted with a solution containing the first and second antibodies to form the ternary complex and the intensity of fluorescence determined and compared to the fluorescence of a standard sample free of said antigen or containing said antigen in a known amount.
Claim 5
Dependent Claim
33 words
A process according to claim 2 wherein the first antibody is bound to a solid carrier that is insoluble in the fluid sample and said second antibody is soluble in the fluid sample.
Claim 6
Dependent Claim
56 words
A process according to claim 4 wherein the fluid sample is simultaneously contacted with said first and second antibodies to form an insoluble ternary complex and the intensity of fluorescence of the ternary complex determined and compared to the fluorescence of a standard sample free of said antigen or containing said antigen in a known amount.
Claim 7
Dependent Claim
33 words
A process according to claim 2 wherein the second antibody is bound to a solid carrier that is insoluble in the fluid sample and said first antibody is soluble in the fluid sample.
Claim 8
Dependent Claim
54 words
A process according to claim 6 wherein the fluid sample is simultaneously contacted with said first and second antibodies to form an insoluble complex and the intensity of fluorescence of the fluid determined and compared to the fluorescence of a standard sample free of said antigen or containing said antigen in a known amount.
Claim 9
Dependent Claim
67 words
A process according to claims 4 or 6 wherein the sample is first contacted with the first antibody to form an antibody:antigen binary complex and then contacted with the second antibody to form the ternary complex and the intensity of the fluorescence of the complex or the fluid determined and compared to a standard sample free of said antigen or containing the antigen in a known amount.
Claim 10
Dependent Claim
28 words
A process according to claims 2, 3, 4, 5, 6, or 7 wherein the fluorescent chromophore is fluorescein and the chromophore capable of absorbing emitted light is rhodamine.
Claim 11
Dependent Claim
22 words
A process according to claim 8 wherein the fluorescent chromophore is fluorescein and the chromophore capable ot absorbing emitted light is rhodamine.
Claim 12
Dependent Claim
68 words
A process according to claim 1 wherein said first antibody is bound to particles insoluble in the fluid sample and said second antibody is bound to particles insoluble in the fluid sample and wherein the sample is contacted with a suspension of the particles for a time sufficient to cause formation of the ternary complex whereby agglomeration of the particles bound to said first and second antibodies occurs.
Claim 13
Dependent Claim
23 words
A process according to claim 11 wherein the size of the particles is in the range of from about 0.2μ to about 10μ.
Claim 14
Dependent Claim
27 words
A process according to claim 12 wherein the particles are selected from the group consisting of particles of latex, silica, glass, cells, polyacrylamide, polymethyl methacrylate and agarose.
Claim 15
Dependent Claim
12 words
A process according to claim 13 wherein the particles are latex particles.
Claim 16
Dependent Claim
28 words
A process according to claims 11, 12, 13, or 14 wherein the particles binding the first antibody are of different color than the particles binding the second antibody.
Claim 17
Dependent Claim
25 words
A process according to claims 12, 13, or 14 wherein the size of the particles is in the range of from about 1.0 to 10μ.
Claim 18
Dependent Claim
22 words
A process according to claim 15 wherein the size of the particles is in the range of from about 1.0 to 10μ.
Claim 19
Dependent Claim
49 words
A process according to claims 11, 12, 13, or 14 wherein the turbidity of the sample after formation of the ternary complex is determined and related to the turbidity of a control sample known to be free of the antigen or to contain a known amount of the antigen.
Claim 20
Dependent Claim
73 words
A process according to claims 11, 12, 13, or 14 wherein a fluorescent chromophore is bound to said first antibody and a chromophore capable of absorbing light at the wavelength emitted by the fluorescent chromopore is bound to the second antibody and wherein, after said contacting, the intensity of fluorescence is determined and compared to the fluorescence of a standard sample free of said antigen or containing said antigen in a known amount.
Claim 21
Dependent Claim
22 words
A process according to claim 19 wherein the fluorescent chromophore is flurescein and the chromophore capable of absorbing emitted light is rhodamine.
Claim 22
Dependent Claim
38 words
A process according to claim 1 wherein an enzyme is bound to the first antibody and a substance is bound to the second antibody whereby the enzyme interacts with the substance to permit detection of the antibody:antigen:antibody complex.
Claim 23
Dependent Claim
17 words
A process according to claim 21 wherein the substance bound to the second antibody is an enzyme.
Claim 24
Dependent Claim
30 words
A process according to claim 22 whereby the first antibody bound enzyme catalyzes a reaction which produces a product required by a reaction catalyzed by the second antibody bound enzyme.
Claim 25
Dependent Claim
29 words
A process according to claim 23 whereby the product of the reaction catalyzed by the first bound enzyme is consumed in the reaction catalyzed by the second bound enzyme.
Claim 26
Dependent Claim
25 words
A process according to claim 23 whereby the product of the reaction catalyzed by the first bound enzyme allosterically interacts with the second bound enzyme.
Claim 27
Dependent Claim
24 words
A process according to claim 25 whereby the product of the reaction catalyzed by the first bound enzyme allosterically activates the second bound enzyme.
Claim 28
Dependent Claim
24 words
A process according to claim 25 whereby the product of the reaction catalyzed by the first bound enzyme allosterically inhibits the second bound enzyme.
Claim 29
Dependent Claim
25 words
A process according to claims 23, 24, 25, 26 or 27 wherein the reaction catalyzed by the second antibody bound enzyme produces a detectable product.
Claim 30
Dependent Claim
21 words
A process according to claim 28 wherein the formation of the detectable product is detected by colorimetry, fluorimetry, luminescence or spectrophotometry.
Claim 31
Dependent Claim
25 words
A process according to claims 23, 24, 25, 26 or 27 wherein the reaction catalyzed by the second antibody bound enzyme consumes a detectable substance.
Claim 32
Dependent Claim
21 words
A process according to claim 30 wherein the consumption of the detectable substance is detected by colorimetry, fluorimetry, luminescence or spectrophotometry.
Claim 33
Dependent Claim
23 words
A process according to claim 21 wherein the substance on the second antibody undergoes a reaction catalyzed by the first antibody bound enzyme.
Claim 34
Dependent Claim
19 words
A process according to claim 32 wherein the reaction produces a substance detectable by colorimetry, fluorimetry, luminescence or spectrophotometry.
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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) | 1987-12-04 | 1988-06-04 | 1988-12-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-12-04 | 1992-06-04 | 1992-12-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-12-04 | 1996-06-04 | 1996-12-04 | $7700.00 | Window Closed / Lapsed |