US4200508A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Method and composition for detecting antigenic substances
Assignee: Individual
•
Filing: 1978-02-09
•
Granted: 1980-04-29
•
Expires: 1998-02-09
Abstract
Abstract A method and composition for detecting an antigenic substance by immunodiffusion using an antigenic substance to be detected labeled by a chromogenic enzyme. According to the method and the detection material of this invention, even a small amount of an antigenic substance can be easily detected.
Inventors
- 👤 Hidematsu Hirai
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 21:41
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/558; Y; Y10; Y10S; Y10S435/00; Y10S435/961
Claims Analysis
Total Claims: 32
Claim 1
Independent Claim
104 words
A method of detecting an antigen by immunodiffusion which comprises: (A) preparing a mixture of a support and an antibody against said antigen; (B) coating said mixture on a plate; (C) forming holes in said coating; (D) applying test sample to said holes in the presence of enzyme-labeled antigen; (E) conducting immunodiffusion to thereby obtain migration of the resulting antigen-antibody complex; (F) contacting said coated plate with a chromophore and a developing agent, whereby a measurable visible indication of the extent of migration is produced by the coupling of said enzyme and said chromphore; and (G) measuring the extent of migration of said complex.
Claim 2
Independent Claim
104 words
A method of detecting an antigen by immunodiffusion which comprises: (A) preparing a mixture of a support and an antibody against said antigen; (B) coating said mixture on a plate; (C) forming holes in said coating; (D) applying test sample to said holes in the presence of enzyme-labeled antigen; (E) conducting immunodiffusion to thereby obtain migration of the resulting antigen-antibody complex; (F) contacting said coated plate with a chromophore and a developing agent, whereby a measurable visible indication of the extent of migration is produced by the coupling of said enzyme and said chromphore; and (G) measuring the extent of migration of said complex.
Claim 3
Dependent Claim
55 words
The method of claim 1, wherein said immunodiffusion is immunoelectrophoresis, further comprising preparing a second mixture of a support and said enzyme-coupled antigen, coating said first-mentioned mixture on a portion of said plate, coating said second mixture on the remaining portion of said plate, and forming said holes in the coating containing said first mixture.
Claim 4
Dependent Claim
25 words
The method of claim 1, wherein said immunodiffusion is single radial diffusion and said enzyme-coupled antigen is applied to said holes with said test sample.
Claim 5
Dependent Claim
15 words
The method of claim 1, wherein the enzyme of said enzyme-labeled antigen is an oxidoreductase.
Claim 6
Dependent Claim
15 words
The method of claim 4, wherein said enzyme is peroxidase, catalase, oxidase, or a dehydrogenase.
Claim 7
Dependent Claim
24 words
The method of claim 1, wherein said support is agar, agarose, gelatin, pectin, silica gel, starch, algin, carrageenin, cross-linked polyacrylamide, or a denatured cellulose.
Claim 8
Dependent Claim
48 words
A structure for use in detecting an antigen by immunoelectrophoresis comprising a plate having coated on a portion thereof a uniform mixture of a support and an antibody against said antigen, and coated on the remaining portion thereof a uniform mixture of a support and an enzyme-labeled antigen.
Claim 9
Dependent Claim
48 words
A structure for use in detecting an antigen by immunoelectrophoresis comprising a plate having coated on a portion thereof a uniform mixture of a support and an antibody against said antigen, and coated on the remaining portion thereof a uniform mixture of a support and an enzyme-labeled antigen.
Claim 10
Dependent Claim
46 words
The method of claim 1, wherein said enzyme-labeled antigen is prepared by coupling a blocking group to the amine group of the enzyme, oxidizing the saccharose moiety of said enzyme to form an aldehyde group, coupling said enzyme to the antigen, and removing said blocking group.
Claim 11
Dependent Claim
11 words
The method of claim 8, wherein said enzyme is an oxidoreductase.
Claim 12
Dependent Claim
15 words
The method of claim 9, wherein said enzyme is peroxidase, catalase, oxidase, or a dehydrogenase.
Claim 13
Dependent Claim
10 words
The method of claim 10, wherein the enzyme is peroxidase.
Claim 14
Dependent Claim
15 words
The method of claim 8, wherein the blocking agent supplying said blocking group is 1-fluoro-2,4-dinitrobenzene.
Claim 15
Dependent Claim
17 words
The method of claim 8, wherein said oxidizing is conducted using sodium periodate as the oxidizing agent.
Claim 16
Dependent Claim
12 words
The method of claim 1, wherein said chromophore is an azo compound.
Claim 17
Dependent Claim
14 words
The method of claim 14, wherein said chromophore is 3,3'-diaminobenzidine, flavin, or a cytochrome.
Claim 18
Dependent Claim
10 words
The method of claim 15, wherein said chromophore is 3,3'-diaminobenzidine.
Claim 19
Dependent Claim
29 words
The method of claim 1, wherein said developing agent is an oxidizing or reducing agent selected from the group consisting of peroxides, oxygen, NADH, FAD, thiols, and metal compounds.
Claim 20
Dependent Claim
15 words
The method of claim 17, wherein said metal compounds are compounds of iron or copper.
Claim 21
Dependent Claim
12 words
The method of claim 17, wherein said developing agent is hydrogen peroxide.
Claim 22
Dependent Claim
51 words
The method of claim 1, wherein said antigen is human α-fetoprotein, human γ-globulin G, or human γ-globulin E; said support is agar or agarose; said enzyme of said enzyme-labeled antigen is peroxidase; said chromophore is 3,3'-diaminobenzidine; said developing agent is hydrogen peroxide; and said immunodiffusion is immunoelectrophoresis or single radial diffusion.
Claim 23
Dependent Claim
18 words
The method of claim 1 wherein the antigen is human α-fetoprotein, human γ-globulin G, or human γ-globulin E.
Claim 24
Dependent Claim
11 words
The method of claim 5 wherein the chromogenic enzyme is peroxidase.
Claim 25
Dependent Claim
13 words
The method of claim 6 wherein the supporting material is agar or agarose.
Claim 26
Dependent Claim
39 words
The method of claim 1 wherein the antigenic is human γ-fetoprotein, human γ-globulin G, or human γ-globulin E; the support is agar or agarose; the enzyme is peroxidase; and the immunodiffusion is an immunoelectrophoresis or a single radial diffusion.
Claim 27
Dependent Claim
40 words
A structure for use in detecting an antigen by immunodiffusion comprising a uniform mixture of a support and an antibody or antiserum against said antigen and a uniform mixture of a support and said antigen labelled by a chromogenic enzyme.
Claim 28
Dependent Claim
40 words
A structure for use in detecting an antigen by immunodiffusion comprising a uniform mixture of a support and an antibody or antiserum against said antigen and a uniform mixture of a support and said antigen labelled by a chromogenic enzyme.
Claim 29
Dependent Claim
30 words
The detection material of claim 25 wherein the antigen is human γ-fetoprotein, human γ-globulin G, or human γ-globulin E; the enzyme is peroxidase; and the support is agar or agarose.
Claim 30
Dependent Claim
30 words
The detection material of claim 7 wherein the antigen is human α-fetoprotein, human γ-globulin G, or human γ-globulin E; the enzyme is peroxidase; and the support is agar or agarose.
Claim 31
Independent Claim
105 words
A method of detecting an antigen by immunodiffusion which comprises: (A) providing a test plate having a coating of a mixture comprising a support and an antibody against said antigen thereon, and holes formed in said coating; (B) applying test sample to said holes in the presence of enzyme-labeled antigen; (C) conducting immunodiffusion to thereby obtain migration of the resulting antigen-antibody complex; (D) contacting said coated plate with a chromophore and a developing agent, whereby a measurable visible indication of the extent of migration is produced by the coupling of said enzyme and said chromophore; and (E) measuring the extent of migration of said complex.
Claim 32
Independent Claim
105 words
A method of detecting an antigen by immunodiffusion which comprises: (A) providing a test plate having a coating of a mixture comprising a support and an antibody against said antigen thereon, and holes formed in said coating; (B) applying test sample to said holes in the presence of enzyme-labeled antigen; (C) conducting immunodiffusion to thereby obtain migration of the resulting antigen-antibody complex; (D) contacting said coated plate with a chromophore and a developing agent, whereby a measurable visible indication of the extent of migration is produced by the coupling of said enzyme and said chromophore; and (E) measuring the extent of migration of said complex.
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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) | 1983-04-29 | 1983-10-29 | 1984-04-29 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-04-29 | 1987-10-29 | 1988-04-29 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-04-29 | 1991-10-29 | 1992-04-29 | $7700.00 | Window Closed / Lapsed |