US4506009A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Heterogeneous immunoassay method
Assignee: University of California
•
Filing: 1982-03-30
•
Granted: 1985-03-19
•
Expires: 2002-03-30
Abstract
Abstract A conjugate useful in determining the amount of antigen or antibody in a liquid sample, said conjugate having a marker, an immunoreactive component (i.e. antigen or antibody) bound to the marker and an insolubilizing binding component which is also bound to the marker. The insolubilizing binding component portion of the conjugate will react with an insolubilizing receptor to form a solid product of conjugate and receptor unless the conjugate reacts with the corresponding antigen or antibody to be analyzed in which event the conjugate will not react with the insolubilizing receptor. The conjugate will be added to a liquid sample containing an unknown amount of, for example, an antibody. A known amount of the corresponding antigen is also added which reacts with both the conjugate and antibody. After the reaction is complete, the liquid sample is contacted with the insolubilizing receptor. Since only the free conjugate reacts with the insolubilizing receptor the amount of antibody originally in the liquid sample can be determined by measuring the activity of the marker in the precipitate.
Inventors
- 👤 Howard M. Lenhoff
- 👤 That T. Ngo
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 02:17
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/975
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
356 words
A heterogeneous immunoassay method for determining the amount of a component to be analyzed, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immaunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said component to be analyzed being dissolved in a liquid sample, which comprises: providing a liquid sample containing a reaction mixture dissolved therein, said reaction mixture consisting essentially of (a) an undetermined amount of said component to be analyzed, (b) a conjugate of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker and which contains a group reactive with an insolubilizing receptor to react with said insolubilizing receptor to form a solid phase in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least about 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being deactivated when said immunobinding partner immunoreacts with said immunoreactive component of said conjugate, and (c) a predetermined amount of said immunobinding partner of the component to be analyzed, said predetermined amount being such that there is a certain amount of unreacted conjugate remaining in said liquid sample; allowing said liquid sample to incubate until the reaction is complete between said immunobinding partner and said component to be analyzed and said immunobinding partner and said conjugate, the reaction product of said immunobinding partner and said conjugate being soluble in said liquid sample; contacting said liquid sample after said reaction is complete with a stoichiometric excess, based on the amount of unreacted conjugate in said liquid sample of said insolubilizing receptor and allowing the reaction between said insolubilizing receptor and said unreacted conjugate to go to completion and form a solid precipitate; separating said solid phase from said liquid sample; and determining the quantity of said component to be analyzed by measuring the activity of the marker in either the solid phase or the liquid sample.
Claim 2
Dependent Claim
356 words
A heterogeneous immunoassay method for determining the amount of a component to be analyzed, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immaunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said component to be analyzed being dissolved in a liquid sample, which comprises: providing a liquid sample containing a reaction mixture dissolved therein, said reaction mixture consisting essentially of (a) an undetermined amount of said component to be analyzed, (b) a conjugate of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker and which contains a group reactive with an insolubilizing receptor to react with said insolubilizing receptor to form a solid phase in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least about 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being deactivated when said immunobinding partner immunoreacts with said immunoreactive component of said conjugate, and (c) a predetermined amount of said immunobinding partner of the component to be analyzed, said predetermined amount being such that there is a certain amount of unreacted conjugate remaining in said liquid sample; allowing said liquid sample to incubate until the reaction is complete between said immunobinding partner and said component to be analyzed and said immunobinding partner and said conjugate, the reaction product of said immunobinding partner and said conjugate being soluble in said liquid sample; contacting said liquid sample after said reaction is complete with a stoichiometric excess, based on the amount of unreacted conjugate in said liquid sample of said insolubilizing receptor and allowing the reaction between said insolubilizing receptor and said unreacted conjugate to go to completion and form a solid precipitate; separating said solid phase from said liquid sample; and determining the quantity of said component to be analyzed by measuring the activity of the marker in either the solid phase or the liquid sample.
Claim 3
Dependent Claim
14 words
A heterogeneous immunoassay method according to claim 1 wherein said marker is an enzyme.
Claim 4
Dependent Claim
32 words
A heterogeneous immunoassay method according to claims 1 or 2 wherein said component to be analyzed is an antigen and said immunobinding partner of said component to be analyzed is an antibody.
Claim 5
Dependent Claim
15 words
A heterogeneous immunoassay method according to claim 1 wherein said insolubilizing binding component is biotin.
Claim 6
Dependent Claim
14 words
A heterogeneous immunoassay method according to claim 4 wherein said insolubilizing receptor is avidin.
Claim 7
Dependent Claim
238 words
A diagnostic kit for determining the amount of a component to be analyzed when said component to be analyzed is dissolved in a liquid sample, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said diagonstic kit consisting essentially of: a separate conjugate of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker, which contains a group reactive with an insolubilizing receptor to react with said receptor to form a solid reaction product which is insoluble in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being capable of being deactivated when said conjugate is contracted with the immunobinding partner of the component to be analyzed; a separate immunobinding partner of the component to be analyzed; and a separate insolubilizing receptor wherein said amount of said immunobinding partner is less than that amount which would react with all of said quantity of said conjugate and said quantity of insolubilizing receptor is a stoichiometric excess of the quantity of unreacted conjugate in said liquid sample.
Claim 8
Dependent Claim
238 words
A diagnostic kit for determining the amount of a component to be analyzed when said component to be analyzed is dissolved in a liquid sample, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said diagonstic kit consisting essentially of: a separate conjugate of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker, which contains a group reactive with an insolubilizing receptor to react with said receptor to form a solid reaction product which is insoluble in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being capable of being deactivated when said conjugate is contracted with the immunobinding partner of the component to be analyzed; a separate immunobinding partner of the component to be analyzed; and a separate insolubilizing receptor wherein said amount of said immunobinding partner is less than that amount which would react with all of said quantity of said conjugate and said quantity of insolubilizing receptor is a stoichiometric excess of the quantity of unreacted conjugate in said liquid sample.
Claim 9
Dependent Claim
13 words
A diagnostic kit according to claim 6 wherein said marker is an enzyme.
Claim 10
Dependent Claim
29 words
A diagnostic kit according to claim 7 wherein said component to be analyzed is an antigen and said immunobinding partner of the component to be analyzed is an antibody.
Claim 11
Dependent Claim
14 words
A diagnostic kit according to claim 6 wherein said insolubilizing binding component is biotin.
Claim 12
Dependent Claim
13 words
A diagnostic kit according to claim 9 wherein said insolubilizing receptor is avidin.
Claim 13
Dependent Claim
178 words
A conjugate for use in determining the amount of a component to be analyzed when said component to be analyzed is dissolved in a liquid sample, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said conjugate consisting essentially of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker and which contains a group reactive with an insolubilizing receptor to react with said receptor to form a solid reaction product which is insoluble in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least about 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being capable of being deactivated when said conjugate is contacted with the immunobinding partner of the component to be analyzed.
Claim 14
Dependent Claim
178 words
A conjugate for use in determining the amount of a component to be analyzed when said component to be analyzed is dissolved in a liquid sample, said component to be analyzed being a component in the reaction of two immunobinding partners, one of said immunobinding partners being an antigen and one of said immunobinding partners being an associated antibody, said conjugate consisting essentially of (i) a marker, (ii) an immunoreactive component bound to said marker and having the same immunochemical properties of said component to be analyzed and (iii) an insolubilizing binding component bound to said marker and which contains a group reactive with an insolubilizing receptor to react with said receptor to form a solid reaction product which is insoluble in said liquid sample, said insolubilizing receptor and said insolubilizing binding component having a binding constant of at least about 10 9 molar, the reactivity of said group of said insolubilizing binding component for said insolubilizing receptor being capable of being deactivated when said conjugate is contacted with the immunobinding partner of the component to be analyzed.
Claim 15
Dependent Claim
12 words
A conjugate according to claim 11 wherein said marker is an enzyme.
Claim 16
Dependent Claim
13 words
A conjugate according to claim 11 wherein said insolubilizing binding component is biotin.
Claim 17
Dependent Claim
13 words
A conjugate according to claim 11 wherein said insolubilizing binding component is avidin.
References Cited (Backward Citations)
US3654090B1
Cited
US3654090A
Cited
US3850752A
Cited
US3839153A
Cited
US3791932A
Cited
US3879262A
Cited
US3935074A
Cited
US4134792A
Cited
US4130462A
Cited
US4298685A
Cited
US4193983A
Cited
US4374925A
Cited
US4288237A
Cited
US4299916A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1988-03-18 | 1988-09-17 | 1989-03-19 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1992-03-18 | 1992-09-17 | 1993-03-19 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1996-03-18 | 1996-09-17 | 1997-03-19 | $7700.00 | Window Closed / Lapsed |