US4289748A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Ultrasensitive enzymatic radioimmunoassay method
Assignee: Government of the United States of America
•
Filing: 1979-05-31
•
Granted: 1981-09-15
•
Expires: 1999-05-31
Abstract
Abstract A method of quantitatively measuring antigen or antibody concentration by selectively coupling an antigen to a solid substrate, coupling an enzyme-conjugated antibody to the antigen and reacting a radioindicator with the enzyme-conjugate producing a radiolabelled product. The product is scintillation counted to yield the measurement. Direct or indirect coupling of the antigen to the substrate can be used. Direct or indirect coupling of the enzyme-conjugate can be used. The preferred radioindicator is adenosine-5'-monophosphate used with alkaline phosphatase. Others are presented.
Inventors
- 👤 Curtis C. Harris
- 👤 Robert Yolken
- 👤 Ih-Chang Hsu
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 03:32
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/534; G01N33/543; G01N33/54306; Y; Y10; Y10S; Y10S436/00; Y10S436/811
Claims Analysis
Total Claims: 21
Claim 1
Independent Claim
82 words
A method of quantitatively measuring the concentration of an immunosystem reactant comprising: (a) preparing a solid substrate; (b) selectively adhering the immunosystem reactant to said substrate; (c) binding an enzyme-conjugated antibody to said reactant; (d) reacting a radiolabelled indicator with said enzyme-conjugated antibody thereby producing a radiolabelled product and an unlabelled product; (e) separating said radiolabelled product(s) from unreacted indicator; and (f) measuring the extent of reaction by radioactive counting said radiolabelled product, said counting yielding a quantitative measurement of the concentration.
Claim 2
Independent Claim
82 words
A method of quantitatively measuring the concentration of an immunosystem reactant comprising: (a) preparing a solid substrate; (b) selectively adhering the immunosystem reactant to said substrate; (c) binding an enzyme-conjugated antibody to said reactant; (d) reacting a radiolabelled indicator with said enzyme-conjugated antibody thereby producing a radiolabelled product and an unlabelled product; (e) separating said radiolabelled product(s) from unreacted indicator; and (f) measuring the extent of reaction by radioactive counting said radiolabelled product, said counting yielding a quantitative measurement of the concentration.
Claim 3
Dependent Claim
16 words
The method according to claim 1 wherein the immunosystem reactant being measured is a specific antigen.
Claim 4
Dependent Claim
102 words
The method according to claim 1 wherein the immunosystem reactant being measured is a single specific antigen-reactive antibody of a particular animal source; step (a) comprises adhering an antigen to said substrate, said antigen being specific to said antibody being measured; step (b) comprises reacting said antibody with said adhered antigen, thereby bending said antibody to said adhered antigen; and step (c) comprises reacting said enzyme-conjugated antibody with said adhered antibody, said enzyme conjugated antibody being selected from the group directed against the immunoglobulin subclass of the animal source of said adhered antibody, thereby binding said enzyme-conjugated antibody to said adhered antibody.
Claim 5
Dependent Claim
37 words
The method according to claim 2 wherein step (a) comprises adhering an antigen-reactive antibody to said substrate, said antigen reactive antibody being specific for said antigen and step (b) comprises reacting said antigen with said adhered antibody.
Claim 6
Dependent Claim
28 words
The method according to claim 2 wherein said enzyme-conjugated antibody is specific for said antigen and step (c) comprises reacting said enzyme-conjugated antibody directly with said adhered antigen.
Claim 7
Dependent Claim
70 words
The method according to claim 2 wherein step (c) comprises reacting an unconjugated antigen-specific antibody to said adhered antigen, said unconjugated antibody being from a different animal source than said adhered antibody; and reacting said enzyme-conjugated antibody with said unconjugated antibody, said enzyme-conjugated antibody being selected from the group directed against the immunoglobulin subclass of the animal source of said unconjugated antibody, thereby binding said enzyme-conjugated antibody to said antigen.
Claim 8
Dependent Claim
37 words
The method according to claim 6 wherein step (a) comprises adhering an antigen-reactive antibody to said substrate, said antigen reactive antibody being specific for said antigen and step (b) comprises reacting said antigen with said adhered antibody.
Claim 9
Independent Claim
24 words
The method according to any one of claims 1, 2, 3, 4, 5, 6, or 7, wherein said radiolabelled indicator is radioactively labelled adenosine-5'-monophosphate.
Claim 10
Dependent Claim
18 words
The method according to claim 8 wherein said adenosine-5'-monophosphate is labelled with H 3 and/or carbon 14 .
Claim 11
Dependent Claim
15 words
The method according to claim 8 wherein said adenosine-5'-monophosphate is labelled with P 32 .
Claim 12
Independent Claim
27 words
The method according to any one of claims 1, 2, 3, 4, 5, 6 or 7, wherein said enzyme-conjugated antibody is an antibody conjugated with alkaline phosphatase.
Claim 13
Dependent Claim
12 words
The method according to claim 11 wherein said antibody is IgG globulin.
Claim 14
Dependent Claim
11 words
The method according to claim 1 wherein step (e) comprises chromatography.
Claim 15
Dependent Claim
12 words
The method according to claim 13 wherein said chromatography comprises liquid chromatography.
Claim 16
Dependent Claim
25 words
The method according to claim 14 wherein said liquid chromatography comprises passing said radiolabelled and said unlabelled products through a selectively adsorbing ion exchange column.
Claim 17
Dependent Claim
20 words
The method according to claim 15, wherein said ion exchange column is packed with cross-linked polymerized dextran ion exchange material.
Claim 18
Dependent Claim
12 words
The method according to claim 14 wherein said chromatography comprises thin-layer chromatography.
Claim 19
Dependent Claim
13 words
The method according to claim 14 wherein said chromatography comprises high pressure chromatography.
Claim 20
Dependent Claim
15 words
The method according to either claim 6 or 7, wherein the antigen is cholera toxin.
Claim 21
Dependent Claim
15 words
The method according to either claim 6 or 7, wherein the antigen is human rotavirus.
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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) | 1984-09-14 | 1985-03-16 | 1985-09-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-09-14 | 1989-03-16 | 1989-09-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-09-14 | 1993-03-16 | 1993-09-15 | $7700.00 | Window Closed / Lapsed |