Patent EP0290269A2 successfully refreshed from Google Patents.
EP0290269A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Immunoassay methods and reagents and methods for producing the latter
Assignee: Cyberfluor Inc
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Filing: 1988-05-06
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Granted: —
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Expires: 2008-05-06
Abstract
Abstract Immunoassay and methods for using fluorescent chelates of lanthanide metal ions in conjunction with immunoreactive bodies to permit fluorescent assay in liquid or dry samples. The assay reagent comprises a residue of an immunoreactive body linked to a residue of a protein or polypeptide. The protein or polypeptide is labelled by substitution with a ligand forming a fluorescent chelate with lanthanide metal ion such as europium. The reagent when used in an immunoassay, binds to the immobilized immunoreactive body and excess is washed away. The sample can be dried for later analysis by a suitable fluoremeter.
Inventors
- 👤 Eleftherios P. Diamandis
- 👤 Alexander J. Lowden
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-02 15:54
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N33/53; G01N33/531; G01N33/532; G01N33/533; Y; Y10; Y10S; Y10S436/00; Y10S436/80; Y10S436/815; Y10S436/817
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
116 words
An immunoassay method wherein immunoreactive bodies immobilized on a solid phase are reacted with complementary immunoreactive bodies characterized in that said complementary immunoassay bodies have connected thereto moieties of a ligand forming with lanthanide metal ion a fluorescent chelate label wherein the metal ion is stably retained, said complementary immunoreactive bodies binding to said immobilized immunoreactive bodies, removing any excess complementary immunoreactive bodies which are not bound to said immobilized bodies, and the fluorescence of the chelate label on bound complementary immunoreactive bodies is measured in the presence of said metal ion stably retained with said chelate, and a value obtained indicative of the quantity of said complementary immunoreactive bodies bound to said immobilized immunoreactive bodies.
Claim 2
Dependent Claim
116 words
An immunoassay method wherein immunoreactive bodies immobilized on a solid phase are reacted with complementary immunoreactive bodies characterized in that said complementary immunoassay bodies have connected thereto moieties of a ligand forming with lanthanide metal ion a fluorescent chelate label wherein the metal ion is stably retained, said complementary immunoreactive bodies binding to said immobilized immunoreactive bodies, removing any excess complementary immunoreactive bodies which are not bound to said immobilized bodies, and the fluorescence of the chelate label on bound complementary immunoreactive bodies is measured in the presence of said metal ion stably retained with said chelate, and a value obtained indicative of the quantity of said complementary immunoreactive bodies bound to said immobilized immunoreactive bodies.
Claim 3
Dependent Claim
20 words
A method according to claim 1 in which the chelate is at least air dried before its fluorescence is measured.
Claim 4
Dependent Claim
28 words
A method according to claim 1 in which the ligand is a moiety of a 4,7-diphenyl-1,10-phenanthroline-2-9,dicarboxylic acid capable of fluorescing in the presence of a lanthanide metal ion.
Claim 5
Dependent Claim
61 words
A method of claim 1, 2 or 3 in which the immunoreactive body immobilized on said solid phase is selected from the group consisting of cortisol, cortisol amine, thyroxine, digoxin, biotin, α-fetoprotein, human chorionic gonadotropin, ferritin, thyrotropin, follitropin, lutropin, thyroxine, binding globulin, growth hormone, prolactin, antibody to cortisol, antibody to α-fetoprotein, antibody to human chorionic gonadotropin and antibody to Rubella virus.
Claim 6
Independent Claim
27 words
A method of any of the preceeding claims in which said lanthanide metal ion is selected from the group consisting of europium, terbium, gadolinium, samarium and dysprosium.
Claim 7
Dependent Claim
35 words
A method of claim 4 in which said immobilized immunoreactive body is α-fetoprotein, said complementary immunoreactive bodies are monoclonal antibodies specific to said α-fetoprotein, said monoclonal antibodies having directly connected thereto said fluorescent chelate labels.
Claim 8
Dependent Claim
36 words
A method of claim 4 in which said immobilized immunoreactive bodies are cortisol molecules, said complementary immunoreactive bodies are monoclonal antibodies specific to said cortisol, said monoclonal antibodies having directly connected thereto said fluorescent chelate labels.
Claim 9
Dependent Claim
43 words
A method of claim 4 in which said immobilized immunoreactive bodies are cortisol molecules, said complementary immunoreactive bodies are antibodies specific to said cortisol molecules, said antibodies being linked by a linking bifunctional reagent to a protein labelled with said fluorescent chelate labels.
Claim 10
Dependent Claim
21 words
A method of claim 8 in which said protein is bovine serum albumin and said linking component is derived from sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate.
Claim 11
Dependent Claim
46 words
A method of claim 4, wherein said immobilized immunoreactive bodies are monoclonal antibodies specific to cortisol which is bound to said solid phase, said complementary immunoreactive bodies are polyclonal antibodies specific to said monoclonal antibodies, said polyclonal antibodies having directly connected thereto said fluorescent chelate labels.
Claim 12
Dependent Claim
58 words
A method of claim 4, wherein said immobilized immunoreactive bodies are selected from the group consisting of human chorionic gonadotropin and α-fetoprotein, said complementary immunoreactive bodies are antibodies specific to the selected immobilized immunoreactive bodies, said antibodies being biotinylated and having bound thereto avidin or streptavidin wherein avidin or streptavidin has connected directly thereto said fluorescent chelate labels.
Claim 13
Dependent Claim
76 words
A method of claim 1, 2, 3 or 4 in which said complementary immunoreactive bodies comprise a two component system, a first component is immunoreactive with said immobilized immunoreactive body, said first component being biotinylated, any excess of said first component being removed, reacting a second component of avidin or streptavidin with said biotinylated first component, said second component being labelled with with moieties of a ligand forming with lanthanide metal ion a fluorescent chelate label.
Claim 14
Dependent Claim
41 words
A reagent for immunoassay characterized in a residue of an immunoreactive body linked to a residue of a protein or polypeptide, said protein or polypeptide being labelled by substitution with moieties of a ligand forming fluorescent chelates with lanthanide metal ions.
Claim 15
Dependent Claim
41 words
A reagent for immunoassay characterized in a residue of an immunoreactive body linked to a residue of a protein or polypeptide, said protein or polypeptide being labelled by substitution with moieties of a ligand forming fluorescent chelates with lanthanide metal ions.
Claim 16
Dependent Claim
67 words
A reagent according to claim 13 wherein the immunoreactive body is a hapten and said residue of a protein or polypeptide comprises a residue selected from the group consisting of residues of bovine serum albumin, thyroglobulin, polylysines of molecular weight 4,000 to 400,000 and lysine copolymers with one or more of alanine, phenylalanine, serine, tyrosine¸ tryptophan and glutamic acid, hemocyanin, myosin, ferritin, catalase and reduced forms thereof.
Claim 17
Dependent Claim
24 words
A reagent according to claim 13 or 14 wherein the body is linked to said molecule through a divalent residue of a bifunctional reagent.
Claim 18
Dependent Claim
50 words
A reagent according to claim 13 or 14 wherein said immunoreactive body is biotinylated and said protein is avidin or streptavidin which is substituted with a plurality of moieties of a ligand forming a fluorescent chelate with lanthanide metal ion exhibiting a Stokes shift of at least about 200 nm.
Claim 19
Dependent Claim
20 words
A reagent according to any one of the preceeding claims wherein the ligand is a moiety of a 4,7-diphenyl-1,10-phenanthroline-2,9-dicarboxylic acid.
Claim 20
Dependent Claim
17 words
A reagent according to any one of the preceeding claims, wherein said lanthanide metal ion is europium.
References Cited (Backward Citations)
No citation records indexed.
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