US4230797A
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Heterogenous specific binding assay employing a coenzyme as label
Assignee: Bayer Corp
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Filing: 1978-04-10
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Granted: 1980-10-28
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Expires: 1998-04-10
Abstract
Abstract An improved heterogeneous specific binding assay method which employs a substance having reactant activity, i.e., a reactant, as a labeling substance in the detection of a ligand in a liquid medium. The method is carried out using reagent means which comprises, as its labeled constituent, a conjugate formed of a specific binding substance coupled to the reactant. The reactant advantageously is an enzymatic reactant such as an enzyme substrate or coenzyme. The activity of the conjugated reactant as a constituent of a predetermined reaction system is utilized as means for monitoring the extent of binding of the labeled constituent in conventional heterogeneous specific binding assay schemes. The presence of a ligand in a liquid medium may be determined following conventional competitive binding manipulative techniques. After the necessary separation of the bound-and free-phases resulting in the specific binding reaction system, the extent of binding of the labeled constituent is determined by contacting either phase with the necessary materials to form the predetermined monitoring reaction system in which the labeling substance is active and assessing reactant activity therein.
Inventors
- 👤 Robert C. Boguslaski
- 👤 Robert J. Carrico
- 👤 James E. Christner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 11:26
CPC Classifications
C; C07; C07D; C07D209/00; C07D209/02; C07D209/44; C07D209/48; C07D237/00; C07D237/26; C07D237/30; C07D237/32; C07D495/00; C07D495/02; C07D495/04; C07H; C07H19/00; C07H19/02; C07H19/04; C07H19/16; C07H19/20; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535; G01N33/536; G01N33/537; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/962; Y10S435/966; Y10S435/968; Y10S435/971; Y10S436/00; Y10S436/805; Y10S436/815
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
165 words
In a heterogeneous specific binding assay method for determining a ligand in a liquid medium, which method comprises the steps of: (a) contacting said liquid medium with reagent means including a labeled conjugate comprising a specific binding substance coupled to a labeling substance, said reagent system producing (1) a bound-phase of the labeled conjugate in which the specific binding substance therein is bound by a specific binding partner thereto and (2) a free-phase of the labeled conjugate in which the specific binding substance therein is not bound by a specific binding partner thereto, (b) separating said bound-phase and said free-phase; and (c) determining said labeling substance in said bound-phase or said free-phase as a function of the amount of said ligand in said liquid medium; the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme, and wherein said labeling substance measuring the extent to which said coenzyme participates in an enzymatic reaction involving an enzyme which requires said coenzyme for activity.
Claim 2
Dependent Claim
165 words
In a heterogeneous specific binding assay method for determining a ligand in a liquid medium, which method comprises the steps of: (a) contacting said liquid medium with reagent means including a labeled conjugate comprising a specific binding substance coupled to a labeling substance, said reagent system producing (1) a bound-phase of the labeled conjugate in which the specific binding substance therein is bound by a specific binding partner thereto and (2) a free-phase of the labeled conjugate in which the specific binding substance therein is not bound by a specific binding partner thereto, (b) separating said bound-phase and said free-phase; and (c) determining said labeling substance in said bound-phase or said free-phase as a function of the amount of said ligand in said liquid medium; the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme, and wherein said labeling substance measuring the extent to which said coenzyme participates in an enzymatic reaction involving an enzyme which requires said coenzyme for activity.
Claim 3
Dependent Claim
36 words
A method as in claim 1 wherein said nucleotide coenzyme is selected from the group consisting of the adenosine phosphates. nicotinamide adenine dinucleotide and reduced forms thereof, and nicotinamide adenine dinucleotide phosphate and reduced forms thereof.
Claim 4
Dependent Claim
19 words
A method as in claim 1 wherein said nucleotide coenzyme is nicotinamide adenine dinucleotide or a reduced form thereof.
Claim 5
Dependent Claim
13 words
A method as in claim 1 wherein said nucleotide coenzyme is adenosine triphosphate.
Claim 6
Dependent Claim
14 words
A method as in claim 1 wherein said nucleotide coenzyme is flavin adenine dinucleotide.
Claim 7
Dependent Claim
12 words
A method as in claim 1 wherein said enzymatic reaction is cyclic.
Claim 8
Dependent Claim
19 words
A method as in claim 6 wherein said coenzyme-active labeling substance is a cycled reagent in said cyclic reaction.
Claim 9
Dependent Claim
12 words
A method as in claim 1 wherein said enzymatic reaction is chemiluminescent.
Claim 10
Dependent Claim
37 words
A method as in claim 1 wherein said ligand is selected from the group consisting of antigens and antibodies thereto; haptens and antibodies thereto; and hormones, vitamins, metabolites, and pharmacological agents, and their receptors and binding substances.
Claim 11
Dependent Claim
14 words
A method as in claim 1 wherein said liquid medium is a biological fluid.
Claim 12
Dependent Claim
72 words
In a reagent means for use in determining a ligand in a liquid medium, which means comprises (i) a labeled conjugate comprising said ligand or a specific binding analog thereof coupled to a labeling substance, and (ii) a specific binding partner of said ligand which binding partner is in a form which is insoluble in said liquid medium, the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme.
Claim 13
Dependent Claim
72 words
In a reagent means for use in determining a ligand in a liquid medium, which means comprises (i) a labeled conjugate comprising said ligand or a specific binding analog thereof coupled to a labeling substance, and (ii) a specific binding partner of said ligand which binding partner is in a form which is insoluble in said liquid medium, the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme.
Claim 14
Dependent Claim
35 words
Means as in claim 11 wherein said nucleotide coenzyme is selected from the group consisting of the adenosine phosphates, nicotinamide adenine dinucleotide and reduced forms thereof, and nicotinamide adenine dinucleotide phosphate and reduced forms thereof.
Claim 15
Dependent Claim
18 words
Means as in claim 11 wherein said nucleotide coenzyme is nicotinamide adenine dinucleotide or a reduced form thereof.
Claim 16
Dependent Claim
12 words
Means as in claim 11 wherein said nucleotide coenzyme is adenosine triphosphate.
Claim 17
Dependent Claim
13 words
Means as in claim 11 wherein said nucleotide coenzyme is flavin adenine dinucleotide.
Claim 18
Dependent Claim
36 words
Means as in claim 11 wherein said ligand is selected from the group consisting of antigens and antibodies thereto; haptens and antibodies thereto; and hormones, vitamins, metabolites, and pharmacological agents, and their receptors and binding substances.
Claim 19
Dependent Claim
27 words
Means as in claim 11 wherein at least one of the components of said means is incorporated with a carrier that is insoluble in said liquid medium.
Claim 20
Dependent Claim
19 words
Means as in claim 17 wherein said carrier is a matrix that is absorbent relative to said liquid medium.
Claim 21
Dependent Claim
71 words
In a reagent means for use in determining a ligand in a liquid medium, which means comprises (i) a labeled conjugate comprising a specific binding partner of said ligand coupled to a labeling substance, and (ii) a specific binding partner of said ligand which binding partner is in a form which is insoluble in said liquid medium, the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme.
Claim 22
Dependent Claim
71 words
In a reagent means for use in determining a ligand in a liquid medium, which means comprises (i) a labeled conjugate comprising a specific binding partner of said ligand coupled to a labeling substance, and (ii) a specific binding partner of said ligand which binding partner is in a form which is insoluble in said liquid medium, the improvement wherein said labeling substance in said labeled conjugate is a nucleotide coenzyme.
Claim 23
Dependent Claim
35 words
Means as in claim 19 wherein said nucleotide coenzyme is selected from the group consisting of the adenosine phosphates, nicotinamide adenine dinucleotide and reduced forms thereof, and nicotinamide adenine dinucleotide phosphates and reduced forms thereof.
Claim 24
Dependent Claim
18 words
Means as in claim 19 wherein said nucleotide coenzyme is nicotinamide adenine dinucleotide or a reduced form thereof.
Claim 25
Dependent Claim
12 words
Means as in claim 19 wherein said nucleotide coenzyme is adenosine triphosphate.
Claim 26
Dependent Claim
13 words
Means as in claim 19 wherein said nucleotide coenzyme is flavin adenine dinucleotide.
Claim 27
Dependent Claim
36 words
Means as in claim 19 wherein said ligand is selected from the group consisting of antigens and antibodies thereto; haptens and antibodies thereto; and hormones, vitamins, metabolites, and pharmacological agents, and their receptors and binding substances.
Claim 28
Dependent Claim
27 words
Means as in claim 19 wherein at least one of the components of said means is incorporated with a carrier that is insoluble in said liquid medium.
Claim 29
Dependent Claim
19 words
Means as in claim 25 wherein said carrier is a matrix that is absorbent relative to said liquid medium.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-10-28 | 1984-04-28 | 1984-10-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-10-28 | 1988-04-28 | 1988-10-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-10-28 | 1992-04-28 | 1992-10-28 | $7700.00 | Window Closed / Lapsed |