Patent EP0457213A2 successfully refreshed from Google Patents.
EP0457213A2
Expired / Lapsed
EP
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Barbiturate assay, tracers, immunogens, antibodies and kit
Assignee: Abbott Laboratories
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Filing: 1991-05-10
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Granted: —
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Expires: 2011-05-10
Abstract
Abstract The present invention is directed to a fluorescence polarization immunoassay for barbiturates, to the various components needed for preparing and carrying out such an assay, and to methods of making these components. Specifically, tracers, immunogens and antibodies are disclosed, as well as methods for preparing them and a reagent kit containing them. The tracers and the immunogens are made from substituted barbiturate compounds. A fluorescein moiety is included in the tracer, while a poly(amino acid) forms a part of the immunogen. The assay is conducted by measuring the degree of polarization retention of plane-polarized light that has been passed through a sample containing antiserum and tracer.
Inventors
- 👤 Maciej Adamczyk
- 👤 Luis A. Cantarero
- 👤 Robert Edward Dubler
- 👤 Patrick J. Jonas
- 👤 Jonathan Grote
- 👤 Jane Ann Nelson
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 16:18
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/52; G01N33/53; G01N33/536; G01N33/542; G01N33/94; G01N33/948
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
127 words
A tracer of the formula: wherein: (1) W is oxygen or sulfur; (2) R¹ is an alkyl, alkenyl, aryl, or alkynyl group having a total of from 1 to 12 carbon atoms, and including from 0 to 1 halogen atoms, arranged in a straight or branched chain, and including up to one aliphatic or aromatic ring structure; (3) R³ is R⁴-Fl; (4) Fl is fluorescein or a fluorescein derivative; and (5) R⁴ is a linking group which: (a) has a total of from 0 to 15 carbon atoms and heteroatoms, arranged in a straight or branched chain, and includes up to one aliphatic or aromatic ring structure, said heteroatoms being O, N, S, P or F; and (b) has a total of from 0 to 5 heteroatoms.
Claim 2
Dependent Claim
41 words
The tracer of Claim 1 wherein: (1) R¹ is an alkyl having a total of from 2 to 7 carbon atoms; and (2) R⁴ is a linking group which has a total of from 0 to 10 carbon atoms and heteroatoms.
Claim 3
Dependent Claim
9 words
The tracer of Claim 1 wherein the tracer is:
Claim 4
Dependent Claim
9 words
The tracer of Claim 1 wherein the tracer is:
Claim 5
Dependent Claim
258 words
A process for determining the presence or amount of barbiturates in biological fluids, comprising the steps of: (a) contacting a sample with a barbiturate antiserum, said barbiturate antiserum containing monoclonal or polyclonal antibodies which have been raised against an immunogen of the following formula: wherein: (1) W is oxygen or sulfur; (2) R¹ is an alkyl, alkenyl, aryl, or alkynyl group having a total of from 1 to 12 carbon atoms, including from 0 to 1 halogen atoms, and including up to one aliphatic or aromatic ring structure; (3) R² is R-Q; (4) Q is a poly(amino acid), a poly(amino acid) derivative or another immunogenically active carrier; and (5) R is a linking group which: (a) has a -CH₂-, a -CH=, a or a -NH- at that end of said linking group which is linked to Q; (b) has a total of from 0 to 20 carbon atoms and heteroatoms, arranged in a straight or branched chain, and includes up to one aliphatic or aromatic ring structure, said heteroatoms being O, N, S, P or F; and (c) has a total of from 0 to 7 heteroatoms; and with tracer compounds according to Claim 1, said tracer compounds being capable of producing a detectable fluorescence polarization response to the presence of the barbiturate antiserum; (b) passing plane polarized light through the resulting solution from step (a) to obtain a fluorescence polarization response; and (c) detecting the fluorescence polarization response of the solution of step (b) as a measure of the presence or amount of barbiturate in the sample.
Claim 6
Dependent Claim
18 words
The process of Claim 5 wherein the antibodies have been raised against an immunogen having the following structure:
Claim 7
Dependent Claim
9 words
The process of Claim 5 wherein the tracer is:
Claim 8
Dependent Claim
12 words
The process of Claim 6 wherein the tracer has the following structure:
Claim 9
Dependent Claim
182 words
A reagent kit for determining the presence or amount of barbiturates in biological fluids which comprises: (a) a tracer of Claim 1; and (b) a monoclonal or polyclonal antibody which has been raised against an immunogen of the formula: wherein: (1) W is oxygen or sulfur; (2) R¹ is an alkyl, alkenyl, aryl, or alkynyl group having a total of from 1 to 12 carbon atoms, including from 0 to 1 halogen atoms, and including up to one aliphatic or aromatic ring structure; (3) R² is R-Q; (4) Q is a poly(amino acid), a poly(amino acid) derivative or another immunogenically active carrier; and (5) R is a linking group which: (a) has a -CH₂-, a -CH=, a or a -NH- at that end of said linking group which is linked to Q; (b) has a total of from 0 to 20 carbon atoms and heteroatoms arranged in a straight or branched chain, and includes up to one aliphatic or aromatic ring structure, said heteroatoms being O, N, S, P or F; and (c) has a total of from 0 to 7 heteroatoms.
Claim 10
Dependent Claim
24 words
The reagent kit of Claim 9 which further comprises an amount of riboflavin binding protein which is effective to reduce fluorescence interference by riboflavin.
Claim 11
Dependent Claim
21 words
The reagent kit of Claims 9 or 10 wherein the antibodies has been raised against an immunogen having the following structure:
Claim 12
Dependent Claim
15 words
The reagent kit of Claims 9 or 10 wherein the tracer has the following structure:
References Cited (Backward Citations)
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