US4687736A
Expired / Lapsed
US
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Detection of biological substances by specific labelling techniques
Assignee: WALES COLLEGE OF MEDICINE THE, University of
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Filing: 1984-10-31
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Granted: 1987-08-18
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Expires: 2004-10-31
Abstract
Abstract This invention relates to the detection of biological substances and is particularly though not exclusively applicable to labelling techniques employing antibodies or other specific binding agents. Thus the invention consists in a method for the detection of a biological substance of interest (optionally in an environment of biological cells) in which a marker substance is linked specifically, directly or indirectly, to the substance of interest, the marker substance being selected to act as a catalyst in a subsequent process of reduction of silver using a developing solution.
Inventors
- 👤 Goeffrey R. Newman
- 👤 Bharat Jasani
- 👤 Edward D. Williams
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 18:21
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/581; Y; Y10; Y10S; Y10S435/00; Y10S435/975; Y10S436/00; Y10S436/827
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
90 words
A method of detecting or quantifying a substance of biological interest in which a peroxidase enzyme marker substance is specifically linked with said substance of interest, the peroxidase is reacted with a visualizing agent selected from the group consisting of (a) diaminobenzidine, (b) 4-chloro-1-naphthol, and (c) amino-9-ethyl-carbazole, the product of the visualizing agent/peroxidase reaction is reacted with a gold salt and with a sulphide salt, and the gold sulphide reaction product is then included as a catalyst in a further reaction with a silver developing solution to precipitate reduced silver.
Claim 2
Independent Claim
90 words
A method of detecting or quantifying a substance of biological interest in which a peroxidase enzyme marker substance is specifically linked with said substance of interest, the peroxidase is reacted with a visualizing agent selected from the group consisting of (a) diaminobenzidine, (b) 4-chloro-1-naphthol, and (c) amino-9-ethyl-carbazole, the product of the visualizing agent/peroxidase reaction is reacted with a gold salt and with a sulphide salt, and the gold sulphide reaction product is then included as a catalyst in a further reaction with a silver developing solution to precipitate reduced silver.
Claim 3
Dependent Claim
14 words
A method according to claim 1, in which the gold salt is gold chloride.
Claim 4
Dependent Claim
14 words
A method according to claim 2, in which the sulphide salt is sodium sulphide.
Claim 5
Dependent Claim
13 words
A method according to claim 3, in which the sodium sulphide is neutralized.
Claim 6
Dependent Claim
13 words
A method according to claim 1, in which the visualizing agent is diaminobenzidine.
Claim 7
Dependent Claim
15 words
A method according to claim 1, in which the silver developing solution includes tungstosilicic acid.
Claim 8
Dependent Claim
24 words
A method according to claim 1, in which the marker substance is capable of forming a specific immunological bond with the substance of interest.
Claim 9
Dependent Claim
25 words
A method according to claim 7, in which the biological substance of interest is an antigen, and the marker substance includes a specific antibody thereto.
Claim 10
Dependent Claim
33 words
A method according to claim 1, in which the substance of interest includes nucleic acid and the marker substance comprises a mucleic acid probe and capable of linking specifically with the nucleic acid.
Claim 11
Dependent Claim
29 words
A method according to claim 1, in which the substance of interest includes a carbohydrate, and the marker substance includes a lectin, capable of specifically linking with the carbohydrate.
Claim 12
Dependent Claim
72 words
A kit for use in detecting a substance of biological interest, comprising a marker substance capable of being specifically linked with the substance of biological interest, and said marker substance including peroxidase enzyme, a visualizing agent for said peroxidase selected from the group consisting of diaminobenzidine, chloro-1-naphthol, and amino-9-ethyl-carbazole, and a visualizing enhancing system comprising a solution of gold salt, a solution of a sulphide salt, and a final silver developing solution.
Claim 13
Dependent Claim
72 words
A kit for use in detecting a substance of biological interest, comprising a marker substance capable of being specifically linked with the substance of biological interest, and said marker substance including peroxidase enzyme, a visualizing agent for said peroxidase selected from the group consisting of diaminobenzidine, chloro-1-naphthol, and amino-9-ethyl-carbazole, and a visualizing enhancing system comprising a solution of gold salt, a solution of a sulphide salt, and a final silver developing solution.
Claim 14
Dependent Claim
14 words
A kit according to claim 11, in which the gold salt is gold chloride.
Claim 15
Dependent Claim
14 words
A kit according to claim 11, in which the sulphide salt is sodium sulphide.
Claim 16
Dependent Claim
13 words
A kit according to claim 11, in which the visualizing agent is diaminobenzidine.
Claim 17
Dependent Claim
24 words
A kit according to claim 11, in which the marker substance is capable of forming a specific immunological bond with the substance of interest.
Claim 18
Dependent Claim
25 words
A kit according to claim 11, in which the biological substance of interest is an antigen, and the marker substance includes a specific antibody thereto.
Claim 19
Dependent Claim
33 words
A kit according to claim 11, in which the substance of interest includes nucleic acid and the marker substance comprises a nucleic acid probe and capable of linking specifically with the nucleic acid.
Claim 20
Dependent Claim
29 words
A kit according to claim 11, in which the substance of interest includes a carbohydrate, and the marker substance includes a lectin, capable of specifically linking with the carbohydrate.
References Cited (Backward Citations)
US4508820A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1990-08-17 | 1991-02-16 | 1991-08-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-08-17 | 1995-02-16 | 1995-08-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-08-17 | 1999-02-16 | 1999-08-18 | $7700.00 | Window Closed / Lapsed |