EP0337082A1
Expired / Lapsed
EP
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Immunoassay utilizing biotin bridge with universal solid phase
Assignee: Bio Rad Laboratories Inc
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Filing: 1989-02-18
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Granted: —
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Expires: 2009-02-18
Abstract
Abstract An immunoassay utilizing a biotin bridge is performed with a solid phase with anti-biotin immobilized on its surface. In accordance with the assay, the solid phase is incubated with a biotinylated binding species, preferably biotinylated antibody, specific for the ligand to be detected. This is then reacted with the test sample, followed by a labeled binding member, either in a sandwich-type or competitive-type binding assay. The label is then detected in accordance with conventional detection techniques. Use of the solid phase with only a biotin-specific antibody attached thereto expands the utility and versatility of a single solid phase for use in the detection of an unlimited variety of ligands.
Inventors
- 👤 Shashidhara H.M. Murthy
- 👤 Susan E. Breglio
- 👤 Vincent E. Phillips
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-08 16:23
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/74
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
104 words
A method for detecting a ligand in a sample, said method comprising: (a) incubating a biotinylated binding species susceptible to immunological binding to said ligand with anti-biotin antibody immobilized on a solid support to form ligand-specific binding sites thereon; (b) incubating said solid support with said sample; (c) incubating said solid support with a species bearing a detectable label, said species being selected from (i) a binding species susceptible to immunological binding to said ligand and (ii) an analog of said ligand; and (d) detecting the presence of said label in a phase selected from (i) said liquid solution and (ii) said solid support.
Claim 2
Independent Claim
104 words
A method for detecting a ligand in a sample, said method comprising: (a) incubating a biotinylated binding species susceptible to immunological binding to said ligand with anti-biotin antibody immobilized on a solid support to form ligand-specific binding sites thereon; (b) incubating said solid support with said sample; (c) incubating said solid support with a species bearing a detectable label, said species being selected from (i) a binding species susceptible to immunological binding to said ligand and (ii) an analog of said ligand; and (d) detecting the presence of said label in a phase selected from (i) said liquid solution and (ii) said solid support.
Claim 3
Dependent Claim
22 words
A method in accordance with claim 1 in which said biotinylated binding species and said binding species of step (c)(i) are antibodies.
Claim 4
Dependent Claim
67 words
A method in accordance in claim 1 in which step (b) comprises binding substantially all ligand in said sample to said biotinylated binding species, step (c) comprises binding substantially all ligand thus bound to a binding species susceptible to immunological binding to said ligand and bearing a detectable label, and step (d) comprises detecting the presence of said label having thus become bound to said solid support.
Claim 5
Dependent Claim
85 words
A method in accordance with claim 1 in which step (c) comprises incubating said solid support with said analog; and step (a) includes controlling the amount of ligand-specific binding sites thus formed on said solid support such that said analog and any ligand present in said sample together exceed said amount, and controlling the amount of said analog incubated with said solid support such that the amount of said analog thus becoming bound is inversely proportional to the amount of ligand present in said sample.
Claim 6
Dependent Claim
25 words
A method in accordance with claim 4 in which step (d) comprises detecting the presence of label having thus become bound to said solid support.
Claim 7
Dependent Claim
49 words
A method in accordance with claim 4 in which said ligand is human chorionic gonadotropin, said biotinylated binding species is a biotinylated antibody specific for the β-chain of human chorionic gonadotropin, and said binding species of step (c)(i) is an antibody specific for the α-chain of human chorionic gonadotropin.
Claim 8
Dependent Claim
61 words
A method in accordance with claim 1 in which said ligand is a hormone selected from the group consisting of human chorionic gonadotropin, luteinizing hormone, thyroid-stimulating hormone and follicle-stimulating hormone, said biotinylated binding species is an antibody specific for the α-chain of said hormone, and said binding species of step (c)(i) is an antibody specific for the β-chain of said hormone.
Claim 9
Dependent Claim
17 words
A method in accordance with claim 1 in which steps (a), (b) and (c) are performed simultaneously.
Claim 10
Dependent Claim
85 words
An assay kit for the detection of a ligand in a sample, said assay kit comprising: (a) a solid phase comprising a solid support having immobilized on the surface thereof anti-biotin antibody: (b) a solution of biotinylated binding species susceptible to immunological binding to said ligand; (c) a labeled species selected from (i) a binding species susceptible to immunological binding to said ligand and (ii) an analog of said ligand with binding specificity to said biotinylated binding species substantially equivalent to that of said ligand.
Claim 11
Dependent Claim
85 words
An assay kit for the detection of a ligand in a sample, said assay kit comprising: (a) a solid phase comprising a solid support having immobilized on the surface thereof anti-biotin antibody: (b) a solution of biotinylated binding species susceptible to immunological binding to said ligand; (c) a labeled species selected from (i) a binding species susceptible to immunological binding to said ligand and (ii) an analog of said ligand with binding specificity to said biotinylated binding species substantially equivalent to that of said ligand.
Claim 12
Dependent Claim
20 words
An assay kit in accordance with claim 9 in which said biotinylated binding species and said labeled species are antibodies.
References Cited (Backward Citations)
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