Patent US4477576A successfully refreshed from Google Patents.
US4477576A
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Antigen assay method and kit
Assignee: Mex Research Associates
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Filing: 1982-07-26
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Granted: 1984-10-16
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Expires: 2002-07-26
Abstract
Abstract A method of determining the concentration of an antigen in a sample solution comprising (a) coating an antigen-protein conjugate onto a solid matrix, (b) conjugating an enzyme to an antibody specific for said antigen, (c) to a known quantity of a solution containing the antibody-enzyme conjugate of (b) adding a specified quantity of a sample containing an unknown amount of the antigen whose content is to be determined, (d) contacting the coated solid matrix of (a) with the solution (c) and incubate so as to effect binding between the antibody and antigen, some of the antigen being that from the sample and some being that on the solid matrix, (e) removing the solid matrix from the solution and washing, (f) immersing the solid matrix in a solution containing a known amount of an enzyme-substrate which is acted upon by the enzyme so as to effect reaction between the enzyme and enzyme-substrate to produce a product, and then separating the solid matrix from the solution of enzyme-substrate, and (g) then measuring the solid matrix and/or the solution of enzyme-substrate with a pre-established standard to indicate the amount of antigen which was in the sample added in (c).
Inventors
- 👤 Alice Deutsch
- 👤 Harvey Brandwein
- 👤 Herbert Platt
- 👤 Dianne M. Hunter
- 👤 Andrew Dubitsky
- 👤 Susan M. Durham
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 13:23
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/94; G01N33/9453; G01N33/53; G01N33/543; G01N33/54366; G01N33/74; G01N33/743; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/964; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/81; Y10S436/815; Y10S436/817; Y10S436/823
Claims Analysis
Total Claims: 31
Claim 1
Independent Claim
60 words
An assay element consisting essentially of (a) a solid matrix, (b) a multiplicity of protein molecules carried on said matrix, (c) a multiplicity of antigen molecules respectively conjugated to said protein molecules, (d) a plurality of monoclonal antibody molecules bound to only some of said antigen molecules, and (e) a plurality of enzyme molecules respectively conjugated to said antibody molecules.
Claim 2
Dependent Claim
60 words
An assay element consisting essentially of (a) a solid matrix, (b) a multiplicity of protein molecules carried on said matrix, (c) a multiplicity of antigen molecules respectively conjugated to said protein molecules, (d) a plurality of monoclonal antibody molecules bound to only some of said antigen molecules, and (e) a plurality of enzyme molecules respectively conjugated to said antibody molecules.
Claim 3
Dependent Claim
17 words
An element according to claim 1, wherein said antigen is at least one of digoxin and digitoxin.
Claim 4
Dependent Claim
11 words
An element according to claim 1, wherein said antigen is progesterone.
Claim 5
Dependent Claim
11 words
An element according to claim 1, wherein said antigen is thyroxine.
Claim 6
Dependent Claim
14 words
An element according to claim 1, wherein said antigen comprises at least one estrogen.
Claim 7
Dependent Claim
11 words
An element according to claim 1, wherein said antigen is dilantin.
Claim 8
Dependent Claim
11 words
An element according to claim 1, wherein said antigen is theophylline.
Claim 9
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a peptide.
Claim 10
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a saccharide.
Claim 11
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a glycoprotein.
Claim 12
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a protein.
Claim 13
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a steroid.
Claim 14
Dependent Claim
12 words
An element according to claim 1, wherein said antigen is a glycolipid.
Claim 15
Dependent Claim
13 words
An element according to claim 1, wherein said antigen is a nucleic acid.
Claim 16
Dependent Claim
19 words
An element according to claim 1, wherein said solid matrix is a plastic or other material for immobilizing antigen.
Claim 17
Dependent Claim
13 words
An element according to claim 1, wherein said solid matrix is a polystyrene.
Claim 18
Dependent Claim
11 words
An element according to claim 1, wherein said protein is albumin.
Claim 19
Dependent Claim
13 words
An element according to claim 1, wherein said protein is bovine serum albumin.
Claim 20
Dependent Claim
12 words
An element according to claim 1, wherein said enzyme is a β-galactosidase.
Claim 21
Dependent Claim
13 words
An element according to claim 1, wherein said enzyme is an alkaline phosphatase.
Claim 22
Dependent Claim
12 words
An element according to claim 1, wherein said enzyme is a peroxidase.
Claim 23
Dependent Claim
12 words
An element according to claim 1, wherein said enzyme is an esterase.
Claim 24
Dependent Claim
102 words
As a mercantile unit, a kit for assaying the antigen content of a biological sample consisting essentially of, in packaged combination, (a) a container containing a solid matrix carrying a known quantity of said antigen, (b) a container containing a solution containing a known quantity of a monoclonal antibody specific for said antigen, said antibody being conjugated to an enzyme, (c) a container containing at least one wash solution, and (d) a container containing a solution containing a known quantity of a substrate capable of being acted upon by the enzyme to produce a measurable change in a property of said substrate.
Claim 25
Dependent Claim
102 words
As a mercantile unit, a kit for assaying the antigen content of a biological sample consisting essentially of, in packaged combination, (a) a container containing a solid matrix carrying a known quantity of said antigen, (b) a container containing a solution containing a known quantity of a monoclonal antibody specific for said antigen, said antibody being conjugated to an enzyme, (c) a container containing at least one wash solution, and (d) a container containing a solution containing a known quantity of a substrate capable of being acted upon by the enzyme to produce a measurable change in a property of said substrate.
Claim 26
Dependent Claim
29 words
A kit according to claim 23, wherein said enzyme-substrate in (d) is one which when acted upon by the enzyme produces a difference in wave length of transmitted light.
Claim 27
Dependent Claim
25 words
A kit according to claim 23, wherein said enzyme-substrate in (d) is one which when acted upon by the enzyme produces a difference in fluorescence.
Claim 28
Dependent Claim
11 words
A kit according to claim 25, wherein said antigen is digoxin.
Claim 29
Dependent Claim
104 words
A process for making an element according to claim 1, comprising (a) coating an antigen-protein conjugate onto the solid substrate, and (b) independently conjugating the enzyme molecules to the molecules of a monoclonal antibody specific for said antigen, (c) contacting the conjugate of (b) with a solution of the antigen and incubating with the coated solid matrix to cause the antibody-enzyme conjugate to bind with antigen, reaching an equilibrium distribution between the antigen carried on the matrix and the antigen in solution, the amount of bound antigen-antibody-enzyme on the substrate being an index of the amount of antigen in the solution of the antigen.
Claim 30
Independent Claim
197 words
A method of determining the concentration of an antigen in a sample solution comprising (a) coating a protein conjugate of said antigen onto a solid matrix, (b) independently conjugating an enzyme to a monoclonal antibody specific for said antigen, (c) incubating a known quantity of a solution containing the antibody-enzyme conjugate of (b) with a specified quantity of a sample containing an unknown amount of the antigen whose content is to be determined, (d) contacting the coated solid matrix of (a) with the solution (c) and incubate so as to effect binding between the antibody and antigen, some of the antigen being that from the sample and some being that on the solid substrate, (e) removing the solid matrix from the solution and washing, (f) immersing the solid matrix in a solution containing a known amount of an enzyme-substrate which is acted upon by the enzyme so as to effect reaction between the enzyme and enzyme-substrate, and then separating the solid matrix from the solution of enzyme-substrate, and (g) then measuring the solid matrix or the solution of enzyme-substrate with a pre-established standard to indicate the amount of antigen which was in the sample added in (c).
Claim 31
Independent Claim
197 words
A method of determining the concentration of an antigen in a sample solution comprising (a) coating a protein conjugate of said antigen onto a solid matrix, (b) independently conjugating an enzyme to a monoclonal antibody specific for said antigen, (c) incubating a known quantity of a solution containing the antibody-enzyme conjugate of (b) with a specified quantity of a sample containing an unknown amount of the antigen whose content is to be determined, (d) contacting the coated solid matrix of (a) with the solution (c) and incubate so as to effect binding between the antibody and antigen, some of the antigen being that from the sample and some being that on the solid substrate, (e) removing the solid matrix from the solution and washing, (f) immersing the solid matrix in a solution containing a known amount of an enzyme-substrate which is acted upon by the enzyme so as to effect reaction between the enzyme and enzyme-substrate, and then separating the solid matrix from the solution of enzyme-substrate, and (g) then measuring the solid matrix or the solution of enzyme-substrate with a pre-established standard to indicate the amount of antigen which was in the sample added in (c).
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