Patent EP0257916A1 successfully refreshed from Google Patents.
US4383031A
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Homogeneous chemiluminescent specific binding assay
Assignee: Bayer Corp
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Filing: 1979-06-21
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Granted: 1983-05-10
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Expires: 1999-06-21
Abstract
Abstract A homogeneous specific binding assay which employs a substance having reactant activity, i.e., a reactant, in a chemiluminescent reaction as a labeling substance in the detection of a ligand in a liquid medium. The assay employs a conjugate formed of a specific binding substance coupled to the chemiluminescent reactant. The activity of the conjugated reactant as a constituent of the chemiluminescent reaction is affected by reaction between the specific binding substance in the conjugate and a specific binding counterpart thereto. The presence of a ligand in a liquid medium may be determined using competitive or displacement binding or sequential saturation techniques wherein the specific binding substance in the conjugate is the ligand or a specific binding analog thereof, or using a direct binding technique wherein the specific binding substance is a specific binding partner of the ligand. The effect of the specific binding reaction on the chemiluminescent activity of the conjugated reactant is related to the presence or amount of the ligand in the liquid medium tested.
Inventors
- 👤 Robert C. Boguslaski
- 👤 Robert J. Carrico
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 00:54
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; C; C07; C07D; C07D209/00; C07D209/02; C07D209/44; C07D209/48; C07D237/00; C07D237/26; C07D237/30; C07D237/32; G01N33/53; G01N33/531; G01N33/532; G01N33/533; G01N33/536; G01N33/542; G01N33/581; Y; Y10; Y10S; Y10S435/00; Y10S435/968; Y10S435/971; Y10S436/00; Y10S436/805; Y10S436/807; Y10S436/808; Y10S436/815; Y10S436/817
Claims Analysis
Total Claims: 51
Claim 1
Independent Claim
133 words
A homogeneous specific binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium (a) with a conjugate comprising said ligand, or a specific binding analog or binding partner thereof, labeled with a substance which is a reactant in a chemiluminescent reaction, and (b) if said conjugate comprises said ligand or analog thereof, also with a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in said chemiluminescent reaction being changed when said conjugate is bound, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) comparing the light produced with that produced following the same procedure using a liquid medium containing a known amount of said ligand.
Claim 2
Dependent Claim
133 words
A homogeneous specific binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium (a) with a conjugate comprising said ligand, or a specific binding analog or binding partner thereof, labeled with a substance which is a reactant in a chemiluminescent reaction, and (b) if said conjugate comprises said ligand or analog thereof, also with a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in said chemiluminescent reaction being changed when said conjugate is bound, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) comparing the light produced with that produced following the same procedure using a liquid medium containing a known amount of said ligand.
Claim 3
Dependent Claim
23 words
The method of claim 1 wherein the rate of said chemiluminescent reaction is measured as a function of said ligand in said medium.
Claim 4
Dependent Claim
26 words
The method of claim 1 wherein the light produced by said chemiluminescent reaction is measured as the total amount of light produced or a portion thereof.
Claim 5
Dependent Claim
22 words
The method of claim 1 wherein the light produced by said chemiluminescent reaction is measured as the peak intensity of light produced.
Claim 6
Dependent Claim
15 words
The method of claim 1 wherein said chemiluminescent label is luciferin, luminol, pyrogallol, or isoluminol.
Claim 7
Dependent Claim
17 words
The method of claim 1 wherein said chemiluminescent label is luminol or isoluminol, or a derivative thereof.
Claim 8
Dependent Claim
11 words
The method of claim 1 wherein said chemiluminescent reaction is enzymatic.
Claim 9
Dependent Claim
21 words
The method of claim 7 wherein said chemiluminescent label is a substrate for an enzyme involved in the enzymatic chemiluminescent reaction.
Claim 10
Dependent Claim
36 words
The method of claim 1 wherein said ligand is selected from the group consisting of antigens and antibodies thereto; haptens and antibodies thereto; and hormones, vitamins, metabolities, and pharmacological agents, and their receptors and binding substances.
Claim 11
Dependent Claim
13 words
The method of claim 1 wherein said liquid medium is a biological fluid.
Claim 12
Dependent Claim
106 words
A homogeneous competitive binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium with (a) a conjugate comprising said ligand or an analog thereof labeled with a chemiluminescent substance, and (b) a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in its chemiluminescent reaction being changed when said conjugate is bound by said binding partner, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) measuring the light produced as a function of said ligand in said liquid medium.
Claim 13
Dependent Claim
106 words
A homogeneous competitive binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium with (a) a conjugate comprising said ligand or an analog thereof labeled with a chemiluminescent substance, and (b) a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in its chemiluminescent reaction being changed when said conjugate is bound by said binding partner, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) measuring the light produced as a function of said ligand in said liquid medium.
Claim 14
Dependent Claim
15 words
The method of claim 11 wherein said chemiluminescent label is luciferin, luminol, pyrogallol, or isoluminol.
Claim 15
Dependent Claim
17 words
The method of claim 11 wherein said chemiluminescent label is luminol or isoluminol, or a derivative thereof.
Claim 16
Dependent Claim
11 words
The method of claim 11 wherein said chemiluminescent reaction is enzymatic.
Claim 17
Dependent Claim
21 words
The method of claim 14 wherein said chemiluminescent label is a substrate for an enzyme involved in the enzymatic chemiluminescent reaction.
Claim 18
Dependent Claim
22 words
The method of claim 11 wherein said ligand is an antigen or a hapten and said binding partner is an antibody therefor.
Claim 19
Dependent Claim
96 words
A homogeneous direct binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium with a conjugate comprising a specific binding partner of said ligand labeled with a chemiluminescent substance, the ability of the chemiluminescent label to participate in its chemiluminescent reaction being changed when said conjugate is bound by said ligand, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) measuring the light produced as a function of said ligand in said liquid medium.
Claim 20
Dependent Claim
96 words
A homogeneous direct binding assay method for determining a ligand in a liquid medium, comprising the steps of: (1) forming a reaction mixture by combining said liquid medium with a conjugate comprising a specific binding partner of said ligand labeled with a chemiluminescent substance, the ability of the chemiluminescent label to participate in its chemiluminescent reaction being changed when said conjugate is bound by said ligand, (2) forming said chemiluminescent reaction in at least a portion of said reaction mixture, and (3) measuring the light produced as a function of said ligand in said liquid medium.
Claim 21
Dependent Claim
15 words
The method of claim 17 wherein said chemiluminescent label is luciferin, luminol, pyrogallol, or isoluminol.
Claim 22
Dependent Claim
17 words
The method of claim 17 wherein said chemiluminescent label is luminol or isoluminol, or a derivative thereof.
Claim 23
Dependent Claim
11 words
The method of claim 17 wherein said chemiluminescent reaction is enzymatic.
Claim 24
Dependent Claim
21 words
The method of claim 20 wherein said chemiluminescent label is a substrate for an enzyme involved in the enzymatic chemiluminescent reaction.
Claim 25
Dependent Claim
20 words
The method of claim 17 wherein said ligand is an antibody and said binding partner is its antigen or hapten.
Claim 26
Dependent Claim
19 words
The method of claim 17 wherein said ligand is an antigen and said binding partner is an antibody therefor.
Claim 27
Dependent Claim
142 words
A homogeneous method of determining the presence of a ligand in a medium suspected of containing said ligand which comprises; bringing together in an aqueous liquid zone: (1) said medium; (2) a chemiluminescent labeled ligand; and (3) a soluble binding partner having sites capable of bonding to said ligand and said chemiluminescent labeled ligand; the concentration of said binding partner being such that there are insufficient binding sites to combine with all of said labeled ligand present in the three component mixture, and wherein binding of said chemiluminescent labeled ligand by said binding partner affects the ability of said labeled ligand to chemiluminesce; and analyzing in said zone, without physically separating free chemiluminescent labeled ligand from chemiluminescent labeled ligand bound to said binding partner, for the effect of said medium on the amount of chemiluminescent labeled ligand bound to said binding partner.
Claim 28
Dependent Claim
142 words
A homogeneous method of determining the presence of a ligand in a medium suspected of containing said ligand which comprises; bringing together in an aqueous liquid zone: (1) said medium; (2) a chemiluminescent labeled ligand; and (3) a soluble binding partner having sites capable of bonding to said ligand and said chemiluminescent labeled ligand; the concentration of said binding partner being such that there are insufficient binding sites to combine with all of said labeled ligand present in the three component mixture, and wherein binding of said chemiluminescent labeled ligand by said binding partner affects the ability of said labeled ligand to chemiluminesce; and analyzing in said zone, without physically separating free chemiluminescent labeled ligand from chemiluminescent labeled ligand bound to said binding partner, for the effect of said medium on the amount of chemiluminescent labeled ligand bound to said binding partner.
Claim 29
Dependent Claim
11 words
A method according to claim 24 wherein said ligand is insulin.
Claim 30
Dependent Claim
15 words
A method according to claim 25 wherein said chemiluminescent labeled ligand is an insulin-luminol conjugate.
Claim 31
Dependent Claim
11 words
A method according to claim 24 wherein said ligand is digoxin.
Claim 32
Dependent Claim
15 words
A method according to claim 27 wherein said chemiluminescent labeled ligand is a digoxin-luminol conjugate.
Claim 33
Dependent Claim
13 words
A method according to claim 24 wherein said binding partner is an antibody.
Claim 34
Dependent Claim
16 words
A method according to claim 24 wherein said ligand is a drug, its analog or metabolite.
Claim 35
Dependent Claim
24 words
A method according to claim 30 wherein said drug is selected from the group consisting of morphine, barbiturates, diphenylhydantoin, and metabolites and analogs thereof.
Claim 36
Dependent Claim
12 words
A method according to claim 24 wherein said ligand is a protein.
Claim 37
Dependent Claim
27 words
A method according to claim 32 wherein said protein is selected from the group consisting of polypeptides, antibodies, hormones, insulin, chorionic gonadotropin, bradykinnin, and thyroid binding globulin.
Claim 38
Dependent Claim
12 words
A method according to claim 24 wherein said ligand is a steroid.
Claim 39
Dependent Claim
12 words
A method according to claim 24 wherein said ligand is a vitamin.
Claim 40
Dependent Claim
28 words
The method of claim 24 wherein said chemiluminescent ligand is a conjugate of a ligand with a chemiluminescent substance selected from the group consisting of luciferin and luminol.
Claim 41
Dependent Claim
73 words
Reagent means for use in a homogeneous specific binding assay for determining a ligand in a liquid medium, comprising (a) a conjugate comprising said ligand or an analog thereof labeled with a substance which is a reactant in a chemiluminescent reaction, and (b) a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in said chemiluminescent reaction being changed when said conjugate is bound by said binding partner.
Claim 42
Dependent Claim
73 words
Reagent means for use in a homogeneous specific binding assay for determining a ligand in a liquid medium, comprising (a) a conjugate comprising said ligand or an analog thereof labeled with a substance which is a reactant in a chemiluminescent reaction, and (b) a specific binding partner of said ligand, the ability of the chemiluminescent label to participate in said chemiluminescent reaction being changed when said conjugate is bound by said binding partner.
Claim 43
Dependent Claim
16 words
The reagent means of claim 37 wherein said chemiluminescent label is luciferin, luminol, pyrogallol, or isoluminol.
Claim 44
Dependent Claim
18 words
The reagent means of claim 37 wherein said chemiluminescent label is luminol or isoluminol, or a derivative thereof.
Claim 45
Dependent Claim
12 words
The reagent means of claim 37 wherein said chemiluminescent reaction is enzymatic.
Claim 46
Dependent Claim
22 words
The reagent means of claim 40 wherein said chemiluminescent label is a substrate for an enzyme involved in the enzymatic chemiluminescent reaction.
Claim 47
Dependent Claim
37 words
The reagent means of claim 37 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 48
Dependent Claim
20 words
The reagent means of claim 37 wherein said conjugate and said specific binding partner are soluble in said liquid medium.
Claim 49
Dependent Claim
19 words
The reagent means of claim 37 wherein said conjugate and said specific binding partner are in a dry form.
Claim 50
Dependent Claim
21 words
The reagent means of claim 37 wherein at least one of the components of said means is incorporated with a carrier.
Claim 51
Dependent Claim
16 words
The reagent means of claim 45 wherein said carrier is adsorbent relative to said liquid medium.
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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) | 1986-05-09 | 1986-11-08 | 1987-05-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-05-09 | 1990-11-08 | 1991-05-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-05-09 | 1994-11-08 | 1995-05-10 | $7700.00 | Window Closed / Lapsed |