US4857455A
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US
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Method for the determination of peroxidase using fluorogenic substrates
Assignee: Syva Co
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Filing: 1982-11-26
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Granted: 1989-08-15
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Expires: 2002-11-26
Abstract
Abstract Novel fluorogenic peroxidase substrates are provided employing novel water soluble activated capped aromatic fluorescent compounds for use as peroxidase substrates. The novel compounds can be used in a wide variety of ways for determination of peroxides, peroxidase, and in assays for analytes employing peroxidase as a label on a member of a specific binding pair.
Inventors
- 👤 Pyare L. Khanna
- 👤 Chiu C. Chang
- 👤 Edwin F. Ullman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 17:34
CPC Classifications
C; C07; C07H; C07H21/00; C12; C12Q; C12Q1/00; C12Q1/26; C12Q1/28; C12Q2326/00; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/968
Claims Analysis
Total Claims: 14
Claim 1
Independent Claim
178 words
In a method for determining the catalystic activity of a perioxidase by measuring the amount of a product produced from a subtrate of said peroxidase, which product provides a fluorometric signal, the improvement which comprises employing as fluorogenic substrate a diaryl organic compound consisting of an aryl fluorescent compound linked by means of a linking group to a carbocyclic aryl group wherein said linking group has a nitrogen or oxygen bonded to a ring carbon atom of said aryl fluorescent compound and an oxygen bonded to a ring carbon atom of said carbocylic aryl group, which diaryl organic compound is non-fluorescent at the excitation wavelength of said aryl fluorescent compound when said aryl fluorescent compound is linked to said carbocylic aryl group, said carbocylic aryl group having an oxy or amino substituent on a ring carbon atom separated by 2 or 4 ring carbon atoms from the ring carbon atom attached to said linking group, which carbocyclic aryl group undergoes perioxidase catalyzed removal as a result of cleavage of said linking group to produce said aryl fluorescent compound.
Claim 2
Dependent Claim
178 words
In a method for determining the catalystic activity of a perioxidase by measuring the amount of a product produced from a subtrate of said peroxidase, which product provides a fluorometric signal, the improvement which comprises employing as fluorogenic substrate a diaryl organic compound consisting of an aryl fluorescent compound linked by means of a linking group to a carbocyclic aryl group wherein said linking group has a nitrogen or oxygen bonded to a ring carbon atom of said aryl fluorescent compound and an oxygen bonded to a ring carbon atom of said carbocylic aryl group, which diaryl organic compound is non-fluorescent at the excitation wavelength of said aryl fluorescent compound when said aryl fluorescent compound is linked to said carbocylic aryl group, said carbocylic aryl group having an oxy or amino substituent on a ring carbon atom separated by 2 or 4 ring carbon atoms from the ring carbon atom attached to said linking group, which carbocyclic aryl group undergoes perioxidase catalyzed removal as a result of cleavage of said linking group to produce said aryl fluorescent compound.
Claim 3
Dependent Claim
14 words
A method according to claim 1, wherein said carbocyclic aryl group is an hydroxyphenyl.
Claim 4
Dependent Claim
13 words
A method according to claim 1, wherein carbocyclic aryl group is an aminophenyl.
Claim 5
Independent Claim
15 words
A method according to any of claims 2 or 3, wherein said group is para.
Claim 6
Dependent Claim
15 words
A method according to claim 1, wherein said carbocyclic aryl group is an oxyaryl group.
Claim 7
Dependent Claim
15 words
A method according to claim 1, wherein said carbocyclic aryl group is an aminoaryl group.
Claim 8
Dependent Claim
13 words
A method according to claim 1, wherein said carbocyclic aryl group is phenyl.
Claim 9
Dependent Claim
244 words
In a method for determining the presence of an analyte in a sanple employing in said method periodidase as a label, wherein said peroxidase is conjugated to a member of a specific binding pair, said pair consisting of ligand and receptor, and said analyte is a member of said pair, said method comprising combining said conjugate, said sample, a source of peroxide, and a substrate for perioxidase, which substrate results in a product producing a detectable signal; and determining the amount of product produced, which amount is related to the amount of analyte in said sample, the improvement which comprises employing as a fluorogenic substrate a diaryl organic compound consisting of an aryl fluorescent compound linked by means of a linking group to a carbocyclic aryl group wherein said linking group has a nitrogen or oxygen bonded to a ring carbon atom of said aryl fluorescent compound and an oxygen bonded to a ring carbon atom of said carbocyclic aryl group, which diaryl organic compound is non-fluorescent at the excitation wavelength of said aryl fluorescent compound when said aryl fluorescent compound is linked to said carbocylic aryl group, said carbocylic aryl group having an oxy or amino substituent on a ring carbon atom separated by 2 or 4 ring carbon atoms from the ring carbon atom attached to said linking group, which carbocylic aryl group undergoes peroxidase catalyzed removal as a result of cleavage of said linking group to produce said aryl fluorescent compound.
Claim 10
Dependent Claim
244 words
In a method for determining the presence of an analyte in a sanple employing in said method periodidase as a label, wherein said peroxidase is conjugated to a member of a specific binding pair, said pair consisting of ligand and receptor, and said analyte is a member of said pair, said method comprising combining said conjugate, said sample, a source of peroxide, and a substrate for perioxidase, which substrate results in a product producing a detectable signal; and determining the amount of product produced, which amount is related to the amount of analyte in said sample, the improvement which comprises employing as a fluorogenic substrate a diaryl organic compound consisting of an aryl fluorescent compound linked by means of a linking group to a carbocyclic aryl group wherein said linking group has a nitrogen or oxygen bonded to a ring carbon atom of said aryl fluorescent compound and an oxygen bonded to a ring carbon atom of said carbocyclic aryl group, which diaryl organic compound is non-fluorescent at the excitation wavelength of said aryl fluorescent compound when said aryl fluorescent compound is linked to said carbocylic aryl group, said carbocylic aryl group having an oxy or amino substituent on a ring carbon atom separated by 2 or 4 ring carbon atoms from the ring carbon atom attached to said linking group, which carbocylic aryl group undergoes peroxidase catalyzed removal as a result of cleavage of said linking group to produce said aryl fluorescent compound.
Claim 11
Dependent Claim
22 words
A method according to claim 8, wherein said source of peroxide is the combination of an oxidase and substrate for said oxidase.
Claim 12
Dependent Claim
15 words
A method according to claim 8, wherein said carbocyclic aryl group is an oxyaryl group.
Claim 13
Dependent Claim
15 words
A method according to claim 8, wherein said carboxyclic aryl group is an aminoaryl group.
Claim 14
Dependent Claim
13 words
A method according to claim 8, wherein said carbocyclic aryl group is phenyl.
References Cited (Backward Citations)
US4279992A
Cited
US4269938A
Cited
US4307188A
Cited
US4318846A
Cited
US4367238A
Cited
US4318981A
Cited
US4439356A
Cited
EP0060518B1
Cited
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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