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EP0034050A2
Expired / Lapsed
EP
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Immunoassay of antigens
Assignee: Fusioni revgroup Pantry Mirror Corp
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Filing: 1981-02-05
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Granted: —
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Expires: 2001-02-05
Abstract
Abstract In the immunoassay of antigens in liquids, a reaction mixture is formed containing the liquid under assay, labelled antigen. and a mixed binding reagent which contains an antigen-binding site and a label-binding site, the two sites being spaced apart in the reagent so that a single molecule of labelled antigen cannot bind to both sites. The label is one whose activity is changed upon binding to a label-binding site, and the amount of antigen in the original liquid sample is determined by measuring the activity of the label in the reaction mixture. A preferred label is a fluorophore. The mixed binding reagent preferably consists of two antibodies linked together.
Inventors
- 👤 David Stuart Smith
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-02 14:03
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/74; G01N33/78; G01N33/53; G01N33/531; G01N33/536; G01N33/542; G01N33/543; G01N33/54306; Y; Y10; Y10S; Y10S436/00; Y10S436/805; Y10S436/815; Y10S436/819; Y10S436/828
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
150 words
.A method of immunoassay of an antigen (Ag) in a liquid sample, which comprises forming a mixture of: a) sample; b) a substance bearing a non-radiosotopic label, which substance is the same as the Ag under assay or is so closely similar as to be bindable by an antibody against the Ag; c) an antibody against the Ag; and detecting the amount of label activity in the mixture and therefrom determining the amount of Ag, characterised in that: component (c) is a mixed binding reagent which comprises at least one first site selectively bindable to the Ag, and at least one second site selectively bindable to the label,the first and second sites being spaced apart so that a single molecule of labelled substance cannot simultaneously become bound to both a first and a second site, and wherein the activity of the label is changed upon binding to the second sites.
Claim 2
Dependent Claim
150 words
.A method of immunoassay of an antigen (Ag) in a liquid sample, which comprises forming a mixture of: a) sample; b) a substance bearing a non-radiosotopic label, which substance is the same as the Ag under assay or is so closely similar as to be bindable by an antibody against the Ag; c) an antibody against the Ag; and detecting the amount of label activity in the mixture and therefrom determining the amount of Ag, characterised in that: component (c) is a mixed binding reagent which comprises at least one first site selectively bindable to the Ag, and at least one second site selectively bindable to the label,the first and second sites being spaced apart so that a single molecule of labelled substance cannot simultaneously become bound to both a first and a second site, and wherein the activity of the label is changed upon binding to the second sites.
Claim 3
Dependent Claim
24 words
A method according to claim 1., characterised in that the label is a fluorophore or a potential fluorophore, and the said activity is fluorescence.
Claim 4
Dependent Claim
39 words
A method according to claim 1 or 2, characterised in that the component (c) reagent comprises an antibody against the Ag and an antibody against the label, the two antibodies being linked together to form a unitary reagent complex.
Claim 5
Dependent Claim
34 words
A method according to claim 3, characterised in that the said two antibodies have been raised in the same animal species and are linked together by an antibody to the immunoglobulin of that species.
Claim 6
Dependent Claim
26 words
A method according to claim 4, characterised in that the two antibodies have been raised in rabbits and the linking antibody has been raised in sheep.
Claim 7
Dependent Claim
18 words
A method according to claim 3, characterised in that the two antibodies are linked together by protein A.
Claim 8
Dependent Claim
18 words
A method according to claim 3, characterised in that the two antibodies are linked to a solid substrate.
Claim 9
Dependent Claim
22 words
A method according to claim 7, characterised in that the solid substrate is finely divided solid particles, each particle bearing both antibodies.
Claim 10
Dependent Claim
15 words
A method according to claim 8, characterised in that the solid particles are polyacrylamide beads.
Claim 11
Dependent Claim
21 words
A method according to claim 3, characterised in that the two antibodies are linked together by a bifunctional chemical bridging group.
Claim 12
Dependent Claim
31 words
TT. A method according to claim 3, characterised in that each antibody is conjugated with a hapten, and the bound haptens are cross-linked to form unitary reagent complexes containing both antibodies.
Claim 13
Dependent Claim
18 words
A method according to claim 11, characterised in that the haptens are cross-linked by antibodies against the hapten.
Claim 14
Independent Claim
19 words
A method according to any preceding claim, characterised in that it is carried out on a discrete manual basis.
Claim 15
Independent Claim
23 words
A method according to any of claims 1 to 12, characterised in that it is carried out on an automated continuous flow basis.
Claim 16
Independent Claim
26 words
A method according to any preceding claim characterised in that the antigen under assay is gentamicin or another aminoglycoside antiobiotic, phenytoin, nortriptyline, or a thyroid hormone.
References Cited (Backward Citations)
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