EP0155224A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Solid-borne complex bearing chromagen responsive functionality for antibody, antigen, receptor, or ligand detection
Assignee: Chromagenics Inc
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Filing: 1985-03-14
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Granted: —
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Expires: 2005-03-14
Abstract
Abstract Disclosed is a reagent useful in the determination of a component of an antibody-antigen or a receptor-receptor binding molecule reaction. The reagent is characterized by the reaction product selected from the group consisting of: (a) a solid support matrix having the antibody linked thereto and a conjugate of the antigen and a chromagenically responsive marker; (b) a solid support matrix having the antigen linked thereto and a conjugate of the antibody and a chromagenically responsive marker; (c) a solid support matrix having the receptor linked thereto and a conjugate of the receptor binding molecule and a chromagenically responsive marker; and (d) a solid support matrix having the receptor binding molecule linked thereto and a conjugate of the receptor and a chromagenically responsive marker. The reagent is especially useful in the detection of hCG in order to ascertain whetherafemale is pregnant. For such embodiment, reagent (a) is composed of the reaction product of nylon having anti-0-hCG linked thereto which is bound to a conjugate of hCG and an enzyme, e.g. horseradish peroxidase, by a carrier molecule such as. for example, Ficoll 70 or bovine serum albumin. Various other non-isotopic immunoassay tests are discussed and disclosed for the reagent.
Inventors
- 👤 David R. Crockford
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 01:03
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/74; G01N33/53; G01N33/531; G01N33/532; G01N33/543
Claims Analysis
Total Claims: 41
Claim 1
Independent Claim
125 words
A reagent for determination of one component of an antibody-antigen reaction or a receptor-receptor binding molecule reaction characterized by the reaction product selected from the group consisting of: (a) a solid support matrix having the antibody linked thereto and a conjugate of the antigen, a carrier molecule, and a chromagenically-responsive marker; (b) a solid support matrix having the antigen linked thereto and a conjugate of the antibody, a carrier molecule, and a ehromagenically-responsive marker; (c) a solid support matrix having the receptor linked thereto and a conjugate of the receptor binding molecule, a carrier molecule, and a chromagenically-responsive marker; and (d) a solid support matrix having the receptor binding molecule linked thereto and a conjugate of the receptor, a carrier molecule, and a chromagenically-responsive marker.
Claim 2
Dependent Claim
125 words
A reagent for determination of one component of an antibody-antigen reaction or a receptor-receptor binding molecule reaction characterized by the reaction product selected from the group consisting of: (a) a solid support matrix having the antibody linked thereto and a conjugate of the antigen, a carrier molecule, and a chromagenically-responsive marker; (b) a solid support matrix having the antigen linked thereto and a conjugate of the antibody, a carrier molecule, and a ehromagenically-responsive marker; (c) a solid support matrix having the receptor linked thereto and a conjugate of the receptor binding molecule, a carrier molecule, and a chromagenically-responsive marker; and (d) a solid support matrix having the receptor binding molecule linked thereto and a conjugate of the receptor, a carrier molecule, and a chromagenically-responsive marker.
Claim 3
Dependent Claim
11 words
The reagent of claim 1 comprising reaction product (a) or (b).
Claim 4
Dependent Claim
17 words
The reagent of claim 1 wherein mixtures of said components are used to form said reaction product.
Claim 5
Dependent Claim
33 words
The reagent of claim 1 wherein said matrix is selected from the group consisting of nylon, polybutadiene, polystyrene, butadiene-styrene copolymers, glass beads, agarose, cellulose, phospholipid bilayer membrane liposome, latex, dextran, and filter paper.
Claim 6
Dependent Claim
11 words
The reagent of claim 4 wherein said matrix is a nylon.
Claim 7
Dependent Claim
12 words
The reagent of claim 2 wherein said antigen is a gonadotropic hormone.
Claim 8
Dependent Claim
41 words
The reagent of claim 1 wherein said marker is selected from the group of an enzyme, a phenol, a heme, an enzyme cofactor, a dye, a chemiluminescence precursor, an enzyme substrate, a fluorochrome, a fluorogen, a fluorescent quencher, and mixtures thereof.
Claim 9
Dependent Claim
11 words
The reagent of claim 7 wherein said marker comprises an enzyme.
Claim 10
Dependent Claim
10 words
The reagent of claim 8 wherein said enzyme is peroxidase.
Claim 11
Dependent Claim
22 words
The reagent of claim 1 wherein the carrier molecule of said conjugate is a sucrose polymer, bovine serum albumin, or mixtures thereof.
Claim 12
Dependent Claim
16 words
The reagent of claim 10 wherein said carrier molecule of said conjugate comprises a sucrose polymer.
Claim 13
Dependent Claim
23 words
The reagent of claim 2 wherein said conjugate contains between about 1 and 15 molecules of marker per molecule of antigen or antibody.
Claim 14
Dependent Claim
23 words
The reagent of claim 2 wherein said conjugate contains between about 4 and 15 molecules of enzyme per molecule of antigen or antibody.
Claim 15
Dependent Claim
62 words
The reagent of claim 1 wherein said reaction product (a) comprises a nylon-6 support matrix having anti-(-human chorionic gonadotropin linked thereto which has been reacted with a conjugate comprising a sucrose polymer carrier molecule containing one or two molecules of human chorionic gonadotropin per molecule of sucrose polymer and between about 4 and 15 molecules of peroxidase per molecule of sucrose polymer.
Claim 16
Independent Claim
58 words
A method for preparing a reagent which is useful in the determination of one component of an antibody-antigen reaction characterized by: (a) covalently linking an antibody to a solid support matrix; (b) separately forming a conjugate with a carrier molecule of said antigen and a chromagenically-responsive marker; and (c) reacting said antibody-modified solid support matrix with said conjugate.
Claim 17
Independent Claim
58 words
A method for preparing a reagent which is useful in the determination of one component of an antibody-antigen reaction characterized by: (a) covalently linking an antibody to a solid support matrix; (b) separately forming a conjugate with a carrier molecule of said antigen and a chromagenically-responsive marker; and (c) reacting said antibody-modified solid support matrix with said conjugate.
Claim 18
Dependent Claim
17 words
The method of claim 15 wherein a mixture of said components is used to prepare said reagent.
Claim 19
Dependent Claim
13 words
The method of claim 15 wherein said solid support matrix comprises a nylon.
Claim 20
Dependent Claim
18 words
The method of claim 15 wherein said antigen and said antibody are interchanged and the reaction steps conducted.
Claim 21
Dependent Claim
12 words
The method of claim 15 wherein said antigen is a gonadotropin hormone.
Claim 22
Dependent Claim
21 words
The method of claim 15 wherein said chromagenically-responsive marker is selected from an enzyme, a phenol, a heme, and mixtures thereof.
Claim 23
Dependent Claim
11 words
The method of claim 20 wherein said marker is an enzyme.
Claim 24
Dependent Claim
10 words
The method of claim 21 wherein said enzyme is peroxidase.
Claim 25
Dependent Claim
27 words
The method of claim 15 wherein said conjugate contains one or two molecules of antigen and between about 4 and 15 molecules of marker per carrier molecule.
Claim 26
Dependent Claim
36 words
The method of claim 23 wherein said solid support matrix is a nylon, said carrier molecule is bovine serum albumin or a sucrose polymer, said marker is an enzyme, and said antigen is human chorionic gonadotropin.
Claim 27
Independent Claim
123 words
A method for the immunochemical determination of a component of an antigen-antibody reaction characterized by: (a) providing a reagent characterized by the reaction product of a solid support matrix having the antibody linked thereto and a conjugate with a carrier molecule of the antigen and a chromagenically-responsive marker; (b) contacting a sample containing the component to be determined with said reagent for a time adequate for the immunochemical reaction to occur; (c) removing said solid support matrix from contact with said sample; (d) adding a chromagen responsive to said marker to said sample; and (e) observing the sample to determine whether a color has appeared which color appearance is indicative of a threshold amount of said component to be determined in said sample.
Claim 28
Independent Claim
123 words
A method for the immunochemical determination of a component of an antigen-antibody reaction characterized by: (a) providing a reagent characterized by the reaction product of a solid support matrix having the antibody linked thereto and a conjugate with a carrier molecule of the antigen and a chromagenically-responsive marker; (b) contacting a sample containing the component to be determined with said reagent for a time adequate for the immunochemical reaction to occur; (c) removing said solid support matrix from contact with said sample; (d) adding a chromagen responsive to said marker to said sample; and (e) observing the sample to determine whether a color has appeared which color appearance is indicative of a threshold amount of said component to be determined in said sample.
Claim 29
Dependent Claim
25 words
The method of claim 23 wherein said solid support matrix is a nylon, said marker is an enzyme, and said antigen is a gonadotropin hormone.
Claim 30
Dependent Claim
12 words
The method of claim 24 wherein said hormone is human chorionic gonadotropin.
Claim 31
Dependent Claim
11 words
The method of claim 25 wherein said enzyme is a peroxidase.
Claim 32
Dependent Claim
16 words
The method of claim 25 wherein said carrier molecule of said conjugate is a sucrose polymer.
Claim 33
Dependent Claim
17 words
The method of claim 25 wherein a mixture of said components is used to prepare said reagent.
Claim 34
Dependent Claim
15 words
The method of claim 30 wherein a mixture of chromagen is used in step (d).
Claim 35
Dependent Claim
17 words
The method of claim 30 wherein step (d) is repeated sequentially with a different chromagen each repeat.
Claim 36
Dependent Claim
49 words
The method of claim 25 for the determination of both estrogen and luteinizing hormone (LH) wherein said reagent (a) is formed by a mixture of estrogen and LH, their corresponding antibodies, and chromagenically-responsive markers; and a multiplicity of chromagens are added in step (d) as a mixture or sequentially.
Claim 37
Dependent Claim
26 words
The method of claim 33 wherein said marker comprises alkaline phosphatase, and step (d) is conducted by sequentially adding p-nitrophenol and endoxyl- phosphate, in either order.
Claim 38
Dependent Claim
84 words
In a reagent for the determination of a component of an antibody-antigen reaction or a receptor-ligand reaction of the type consisting of one component in an insolubilized form and the other component linked to an enzyme, the improvement wherein said reagent comprises: (a) said one component in an insolubilized form; (b) said other component provided in a conjugate of a carrier molecule bearing said second component and between about 4 and 15 molecules of a chromagenically-rsponsive marker molecule per molecule of said second component.
Claim 39
Dependent Claim
84 words
In a reagent for the determination of a component of an antibody-antigen reaction or a receptor-ligand reaction of the type consisting of one component in an insolubilized form and the other component linked to an enzyme, the improvement wherein said reagent comprises: (a) said one component in an insolubilized form; (b) said other component provided in a conjugate of a carrier molecule bearing said second component and between about 4 and 15 molecules of a chromagenically-rsponsive marker molecule per molecule of said second component.
Claim 40
Dependent Claim
13 words
The improved reagent of claim 35 wherein said chromagenically-responsive marker is an enzyme.
Claim 41
Dependent Claim
25 words
The improved reagent of claim 35 wherein said first component is an antibody or receptor and said second component is an antigen or ligand, respectively.
References Cited (Backward Citations)
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