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EP0345776A2
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Assays utilizing sensitizer-induced production of detectable signals
Assignee: Nichols Institute Diagnostics
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Filing: 1989-06-08
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Granted: —
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Expires: 2009-06-08
Abstract
Abstract Specific binding assays are disclosed which utilize a sensitizer as a label. Such sensitizers include any moiety which, when stimulated by "excitation" with radiation of one or more wavelengths or other chemical or physical stimulus (e.g., electron transfer, electrolysis, electroluminescence or energy transfer), will achieve an excited state which (a) upon interaction with molecular oxygen will produce singlet molecular oxygen, or (b) upon interaction with a leucodye will assume a reduced form which can then be returned to its original unexcited state by interaction with molecular oxygen resulting in the production of hydrogen peroxide. Either interaction with the excited sensitizer will, with the addition of other reagents, produce a detectable signal.
Inventors
- 👤 Frank Mccapra
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 02:54
CPC Classifications
C; C07; C07H; C07H21/00; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N33/583; Y; Y10; Y10S; Y10S436/00; Y10S436/905; Y10T; Y10T436/00; Y10T436/14; Y10T436/142222; Y10T436/143333
Claims Analysis
Total Claims: 39
Claim 1
Independent Claim
185 words
In a specific binding assay for the presence of an analyte in a sample wherein (i) said assay utilizes a sensitizer conjugate comprising a sensitizer attached to a specific binding material and (ii) the presence of analyte in said sample is proportional to the formation of one or more specific binding reaction products containing said sensitizer conjugate, said assay comprising: allowing under suitable conditions substantial formation of said one or more specific binding reaction products containing said sensitizer conjugate; allowing under suitable conditions (A) excitation of (i) sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, and (B) reaction of (i) excited sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) excited sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, with molecular oxygen to produce singlet molecular oxygen which singlet molecular oxygen suitable for reaction to produce a detectable signal; and measuring said detectable signal.
Claim 2
Dependent Claim
185 words
In a specific binding assay for the presence of an analyte in a sample wherein (i) said assay utilizes a sensitizer conjugate comprising a sensitizer attached to a specific binding material and (ii) the presence of analyte in said sample is proportional to the formation of one or more specific binding reaction products containing said sensitizer conjugate, said assay comprising: allowing under suitable conditions substantial formation of said one or more specific binding reaction products containing said sensitizer conjugate; allowing under suitable conditions (A) excitation of (i) sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, and (B) reaction of (i) excited sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) excited sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, with molecular oxygen to produce singlet molecular oxygen which singlet molecular oxygen suitable for reaction to produce a detectable signal; and measuring said detectable signal.
Claim 3
Dependent Claim
34 words
The specific binding assay of claim 1 wherein said specific binding material is capable of specifically binding with the analyte and said one or more specific binding reaction products is an analyte-sensitizer conjugate complex.
Claim 4
Dependent Claim
57 words
The specific binding assay of claim 1 wherein said assay further utilizes a reactant which is capable of specifically binding (i) with said analyte to form an analyte-reactant complex and (ii) with said specific binding material to form a sensitizer conjugate-reactant complex, and wherein said one or more specific binding reaction products is said sensitizer conjugate-reactant complex.
Claim 5
Dependent Claim
55 words
The specific binding assay of claim 1 wherein said specific binding material is capable of specifically binding with said analyte and said assay further utilizes a reactant capable of specifically binding with said analyte to form a reactant-analyte-sensitizer conjugate complex, and wherein said one or more specific binding reaction products is said reactant-analyte-sensitizer conjugate complex.
Claim 6
Dependent Claim
38 words
The specific binding assay of any of the claims 1 to 4 wherein said sensitizer is selected from the group consisting of a dye, a porphyrin, a metalloporphyrin, an aromatic hydrocarbon, a heterocyclic compound and a flavin derivative.
Claim 7
Dependent Claim
15 words
The specific binding assay of claim 5 wherein said sensitizer is pyrene or methylene blue.
Claim 8
Dependent Claim
27 words
The specific binding assay of claim 5 wherein said porphyrin is protoporphyrin dimethyl ester, protoporphyrin disodium salt, methyl pyrroporphine ethyl ester, methyl pyrroporphine, tetraphenylporphine, co-porphyrin or hematoporphyrin.
Claim 9
Dependent Claim
15 words
The specific binding assay of claim 5 wherein said sensitizer is phthalocyanine, hemin or rhodamine.
Claim 10
Dependent Claim
36 words
The specific binding assay of any of the claims 1 to 8, wherein said singlet molecular oxygen reacts with an olefin to produce a moiety selected from the group consisting of a dioxetan and a peroxide.
Claim 11
Dependent Claim
21 words
The specific binding assay of claim 9 wherein said moiety decays upon heating to produce said detectable signal as a photon.
Claim 12
Dependent Claim
27 words
The specific binding assay of claim 9 wherein said peroxide oxidizes a chromogen to produce said detectable signal as a color change or fluorescence by said chromogen.
Claim 13
Dependent Claim
14 words
The specific binding assay of claim 9 wherein said olefin is a substituted olefin.
Claim 14
Dependent Claim
22 words
The specific binding assay of claim 12 wherein said substituted olefin has at least one substitution which is an electron donating group.
Claim 15
Dependent Claim
28 words
The specific binding assay of claim 12 wherein at least two of the substitutions of said substituted olefin are joined to form a ringed moiety which is fluorescent.
Claim 16
Dependent Claim
23 words
The specific binding assay of claim 14 wherein said substituted olefin has the formula: and said ringed moiety is an N-methyl acridan moiety.
Claim 17
Dependent Claim
173 words
In a specific binding assay for the presence of an analyte in a sample wherein (i) said assay utilizes a sensitizer conjugate comprising a sensitizer attached to a specific binding material and (ii) the presence of analyte in said sample is proportional to the formation of one or more specific binding reaction products containing said sensitizer conjugate, said assay comprising: allowing under suitable conditions substantial formation of said one or more specific binding reaction products containing said sensitizer conjugate; allowing under suitable conditions (A) excitation of (i) sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, and (B) reaction of (i) excited sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) excited sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, with a leucodye to produce a detectable signal; and measuring said detectable signal.
Claim 18
Dependent Claim
173 words
In a specific binding assay for the presence of an analyte in a sample wherein (i) said assay utilizes a sensitizer conjugate comprising a sensitizer attached to a specific binding material and (ii) the presence of analyte in said sample is proportional to the formation of one or more specific binding reaction products containing said sensitizer conjugate, said assay comprising: allowing under suitable conditions substantial formation of said one or more specific binding reaction products containing said sensitizer conjugate; allowing under suitable conditions (A) excitation of (i) sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, and (B) reaction of (i) excited sensitizer contained in said one or more specific binding reaction products containing said sensitizer conjugate, or (ii) excited sensitizer contained in sensitizer conjugate not contained in said one or more specific binding reaction products, with a leucodye to produce a detectable signal; and measuring said detectable signal.
Claim 19
Dependent Claim
20 words
The specific binding assay of claim 16 further characterized by the features of any of the claims 2 to 8.
Claim 20
Dependent Claim
28 words
The specific binding assay of claim 16 or 17 wherein said leucodye is oxidized to produce said detectable signal as a color change or fluorescence by said leucodye.
Claim 21
Dependent Claim
31 words
The specific binding assay of claim 16 or 17 wherein the excited sensitizer is reduced by said leucodye and is subsequently oxidized by reaction with molecular oxygen to produce hydrogen peroxide.
Claim 22
Dependent Claim
28 words
The specific binding assay of claim 19 wherein said hydrogen peroxide oxidizes a chromogen to produce said detectable signal as a color change or fluorescence by said chromogen.
Claim 23
Dependent Claim
23 words
The specific binding assay of claim 19 wherein said hydrogen peroxide oxidizes a chemiluminescent moiety to produce said detectable signal as a photon.
Claim 24
Dependent Claim
40 words
As an article of manufacture, a specific binding assay kit comprising: a first vial containing a sensitizer conjugate, said sensitizer conjugate comprising a sensitizer attached to a specific binding material; and a second vial containing an olefin or a leucodye.
Claim 25
Dependent Claim
40 words
As an article of manufacture, a specific binding assay kit comprising: a first vial containing a sensitizer conjugate, said sensitizer conjugate comprising a sensitizer attached to a specific binding material; and a second vial containing an olefin or a leucodye.
Claim 26
Dependent Claim
30 words
The article of manufacture of claim 22 wherein said sensitizer is selected from the group consisting of a dye, a porphyrin, a metalloporphyrin, an aromatic hydrocarbon and a flavin derivative.
Claim 27
Dependent Claim
16 words
The article of manufacture of claim 22 or 23 wherein said second vial contains an olefin.
Claim 28
Dependent Claim
29 words
The article of manufacture of claim 24 wherein said olefin is a substituted olefin, preferably a substituted olefin having at least one substitution which is an electron donating group.
Claim 29
Dependent Claim
28 words
The article of manufacture of claim 25 wherein at least two of the substitions of said substituted olefin are joined to form a ringed moiety which is fluorescent.
Claim 30
Dependent Claim
23 words
The article of manufacture of claim 26 wherein said substituted olefin has the formula: and said ringed moiety is an N-methyl acridan moiety.
Claim 31
Dependent Claim
14 words
The article of manufacture of claim 22 wherein said second vial contains a leucodye.
Claim 32
Dependent Claim
12 words
A sensitizer conjugate comprising a sensitizer attached to a specific binding material.
Claim 33
Dependent Claim
12 words
A sensitizer conjugate comprising a sensitizer attached to a specific binding material.
Claim 34
Dependent Claim
21 words
The sensitizer conjugate of claim 26 wherein the sensitizer is a compound according to any of the claims 5 to 8.
Claim 35
Dependent Claim
22 words
The sensitizer conjugate of claim 29 or 30 wherein the specific binding material binds specifically to either A, T, C or G.
Claim 36
Dependent Claim
19 words
The sensitizer conjugate of claim 29 or 30 wherein the specific binding material binds specifically to a polynucleotide segment.
Claim 37
Dependent Claim
21 words
The sensitizer conjugate of claim 29 or 30, wherein the specific binding material binds specifically to protein or a protein conjugate.
Claim 38
Dependent Claim
20 words
The sensitizer conjugate of claim 29 or 30 wherein the specific material is an antibody or comprises an antibody fragment.
Claim 39
Dependent Claim
94 words
The specific binding of assay of claims 1 or 16 wherein said sensitizer conjugate comprises a sensitizer attached to a first single-stranded polynucleotide segment and said assay further utilizes a chemiluminescent moiety conjugate comprising a chemiluminescent moiety attached to a second single-stranded polynucleotide segment, said first and second polynucleotide segments being complementary to mutually exclusive portions of a target single-stranded polynucleotide such that upon hybridization with a target single-stranded polynucleotide the chemiluminescent moiety and the sensitizer are close enough to each other to permit energy transfer from the chemiluminescent moiety to excite the sensitizer.
References Cited (Backward Citations)
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