Patent US5340716A successfully refreshed from Google Patents.
US5340716A
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Assay method utilizing photoactivated chemiluminescent label
Assignee: Siemens Healthcare Diagnostics Products GmbH
•
Filing: 1991-06-20
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Granted: 1994-08-23
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Expires: 2011-06-20
Abstract
Abstract Methods are disclosed for determining an analyte in a medium suspected of containing the analyte. One method comprises providing (1) a medium suspected of containing the analyte, (2) a label reagent comprising a first specific binding pair (sbp) member associated with a photochemically activatable chemiluminescent compound wherein the first sbp member is capable of binding to the analyte or to a second sbp member to form a complex related to the presence of the analyte. The method further comprises photochemically activating the chemiluminescent compound. The amount of luminescence generated by the chemiluminescent compound is detected. The amount thereof is related to the amount of analyte in the medium. Compositions and kits are also disclosed.
Inventors
- 👤 Edwin F. Ullman
- 👤 Hrair Kirakossian
- 👤 John S. Pease
- 👤 Yuri Daniloff
- 👤 Daniel B. Wagner
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 12:17
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/582; G01N33/53; G01N33/569; G01N33/56966; G01N33/56977; G01N33/585
Claims Analysis
Total Claims: 93
Claim 1
Independent Claim
96 words
A method for determining an analyte, said method comprising: providing in combination (1) a medium suspected of containing said analyte, (2) a label reagent comprising a first specific binding pair (sbp) member associated with a photochemically activatable chemiluminescent compound (PACC) wherein said first sbp member is capable of binding to said analyte or to a second sbp member to form a complex related to the presence of said analyte, photochemically activating said PACC, and detecting the amount of luminescence generated by said PACC, the amount thereof being related to the amount of analyte in said medium.
Claim 2
Independent Claim
96 words
A method for determining an analyte, said method comprising: providing in combination (1) a medium suspected of containing said analyte, (2) a label reagent comprising a first specific binding pair (sbp) member associated with a photochemically activatable chemiluminescent compound (PACC) wherein said first sbp member is capable of binding to said analyte or to a second sbp member to form a complex related to the presence of said analyte, photochemically activating said PACC, and detecting the amount of luminescence generated by said PACC, the amount thereof being related to the amount of analyte in said medium.
Claim 3
Dependent Claim
22 words
The method of claim 1 which is a homogenous method and wherein said photochemical activation comprises irradiation of said medium with light.
Claim 4
Dependent Claim
22 words
The method of claim 1 which is a heterogenous method and wherein said photochemical activation comprises excitation of said PACC with light.
Claim 5
Dependent Claim
17 words
The method of claim 1 wherein said photochemical activation comprises reaction of said PACC with singlet oxygen.
Claim 6
Dependent Claim
16 words
The method of claim 4 wherein said singlet oxygen is generated by activation of a photosensitizer.
Claim 7
Dependent Claim
11 words
The method of claim 5 wherein said photosensitizer is a dye.
Claim 8
Dependent Claim
13 words
The method of claim 1 wherein said detecting comprises detecting said luminescence directly.
Claim 9
Dependent Claim
14 words
The method of claim 1 wherein said detecting comprises employing a fluorescent energy acceptor.
Claim 10
Dependent Claim
30 words
The method of claim 8 wherein said fluorescent energy acceptor is bound to, or is capable of becoming bound to, an sbp member other than that of said label reagent.
Claim 11
Dependent Claim
29 words
The method of claim 1 wherein a surface is employed and an energy acceptor is employed that is bound to, or is capable of becoming bound to, said surface.
Claim 12
Dependent Claim
26 words
The method of claim 1 wherein said analyte and said first sbp member are each independently selected from the group consisting of ligands, receptors, and polynucleotides.
Claim 13
Dependent Claim
25 words
The method of claim 1 wherein said PACC contains an olefin group and one or more electron donating substituents in conjugation with said olefin group.
Claim 14
Dependent Claim
23 words
The method of claim 1 wherein said PACC is selected from the group consisting of 9-alkylidene acridans, enol ethers, 9-alkylidene xanthanes and enamines.
Claim 15
Dependent Claim
11 words
The method of claim 1 wherein said analyte is a polynucleotide.
Claim 16
Dependent Claim
13 words
The method of claim 1 wherein said first sbp member is an antibody.
Claim 17
Dependent Claim
15 words
The method of claim 1 wherein said second sbp member is analogous to said analyte.
Claim 18
Dependent Claim
24 words
The method of claim 1 wherein said second sbp member is analogous to or complementary to said analyte or to a third sbp member.
Claim 19
Dependent Claim
15 words
The method of claim 17 wherein said third sbp member is bound to a support.
Claim 20
Dependent Claim
13 words
The method of claim 1 wherein said complex becomes bound to a support.
Claim 21
Dependent Claim
15 words
The method of claim 1 wherein said second sbp member is bound to a support.
Claim 22
Independent Claim
101 words
A method for determining an analyte, said method comprising: combining in an assay medium (1) a medium suspected of containing an analyte, and (2) a label reagent comprising a member of a specific binding pair (sbp member) bound to a photochemically activatable chemiluminescent compound (PACC) under conditions wherein an sbp member complex involving said label reagent is formed in relation to the presence of analyte in said medium, irradiating said assay medium with light to activate said PACC, and examining said assay medium for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 23
Independent Claim
101 words
A method for determining an analyte, said method comprising: combining in an assay medium (1) a medium suspected of containing an analyte, and (2) a label reagent comprising a member of a specific binding pair (sbp member) bound to a photochemically activatable chemiluminescent compound (PACC) under conditions wherein an sbp member complex involving said label reagent is formed in relation to the presence of analyte in said medium, irradiating said assay medium with light to activate said PACC, and examining said assay medium for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 24
Dependent Claim
26 words
The method of claim 21 wherein an energy acceptor is included in said medium and energy from said PACC is capable of activating said energy acceptor.
Claim 25
Dependent Claim
21 words
The method of claim 21 wherein said medium is irradiated with light having a wavelength between 500 nm and 950 nm.
Claim 26
Dependent Claim
18 words
The method of claim 21 wherein said assay medium comprises a second sbp member other than said analyte.
Claim 27
Dependent Claim
11 words
The method of claim 21 wherein said energy acceptor is fluorescent.
Claim 28
Dependent Claim
32 words
The method of claim 25 wherein said fluorescent energy acceptor is bound to, or is capable of becoming bound to, a second sbp member or to a surface included in said medium.
Claim 29
Dependent Claim
25 words
The method of claim 21 wherein said analyte and said sbp member are each independently selected from the group consisting of ligands, receptors, and polynucleotides.
Claim 30
Dependent Claim
25 words
The method of claim 21 wherein said PACC contains an olefin group and one or more electron donating substituents in conjugation with said olefin group.
Claim 31
Dependent Claim
23 words
The method of claim 21 wherein said PACC is selected from the group consisting of 9-alkylidene acridans, enol ethers, 9-alkylidene xanthanes and enamines.
Claim 32
Dependent Claim
14 words
The method of claim 21 wherein said sbp member is complementary to said analyte.
Claim 33
Dependent Claim
18 words
The method of claim 21 wherein said assay medium comprises a second sbp member complementary to said analyte.
Claim 34
Dependent Claim
44 words
The method of claim 31 wherein said second sbp member is bound to, or is capable of becoming bound to, a support and said support is combined with said assay medium prior to, simultaneously with, or subsequent to the addition of said label reagent.
Claim 35
Dependent Claim
31 words
The method of claim 21 wherein said sbp member is analogous to said analyte and said assay medium comprises a second sbp member complementary to said analyte and said sbp member.
Claim 36
Dependent Claim
15 words
The method of claim 33 wherein said second sbp member is bound to a support.
Claim 37
Dependent Claim
29 words
The method of claim 21 wherein said sbp member is complementary to a second sbp member complementary to said analyte and said assay medium comprises said second sbp member.
Claim 38
Dependent Claim
15 words
The method of claim 35 wherein said second sbp member is bound to a support.
Claim 39
Dependent Claim
24 words
The method of claim 21 wherein said second sbp member is analogous to or complementary to said analyte or to a third sbp member.
Claim 40
Dependent Claim
37 words
The method of claim 37 wherein said third sbp member is bound to a support and said support is combined with said assay medium prior to, simultaneously with, or subsequent to the addition of said label reagent.
Claim 41
Independent Claim
142 words
A method for determining an analyte, said method comprising: combining in an assay medium either simultaneously or wholly or partially sequentially (1) a medium suspected of containing an analyte, (2) a label reagent comprising a first member of a specific binding pair (sbp member) bound to a compound capable of chemiluminescence upon reaction with singlet oxygen ("said compound"), and (3) an insolubilized reagent comprising a second sbp member under conditions wherein an sbp member complex involving said label reagent and said insolubilized reagent is formed in relation to the presence of analyte in said medium, separating said assay medium and said insolubilized reagent, activating said compound in said medium or on said insolubilized reagent with light, and examining said assay medium or said reagent for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 42
Independent Claim
142 words
A method for determining an analyte, said method comprising: combining in an assay medium either simultaneously or wholly or partially sequentially (1) a medium suspected of containing an analyte, (2) a label reagent comprising a first member of a specific binding pair (sbp member) bound to a compound capable of chemiluminescence upon reaction with singlet oxygen ("said compound"), and (3) an insolubilized reagent comprising a second sbp member under conditions wherein an sbp member complex involving said label reagent and said insolubilized reagent is formed in relation to the presence of analyte in said medium, separating said assay medium and said insolubilized reagent, activating said compound in said medium or on said insolubilized reagent with light, and examining said assay medium or said reagent for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 43
Dependent Claim
24 words
The method of claim 39 wherein said assay medium is irradiated with light of wavelength between 500 and 950 nm to activate said compound.
Claim 44
Dependent Claim
16 words
The method of claim 39 wherein said singlet oxygen is generated by irradiation of a dye.
Claim 45
Dependent Claim
22 words
The method of claim 41 wherein said dye is selected from the group consisting of methylene blue, rose bengal, porphyrins, and phthalocyanines.
Claim 46
Dependent Claim
25 words
The method of claim 39 wherein said analyte and said sbp members are each independently selected from the group consisting of ligands, receptors, and polynucleotides.
Claim 47
Dependent Claim
25 words
The method of claim 39 wherein said compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin group.
Claim 48
Dependent Claim
23 words
The method of claim 39 wherein said compound is selected from the group consisting of 9-alkylidene acridans, enol ethers, 9-alkylidene xanthanes and enamines.
Claim 49
Dependent Claim
15 words
The method of claim 39 wherein said first sbp member is complementary to said analyte.
Claim 50
Dependent Claim
16 words
The method of claim 39 wherein said a second sbp member is complementary to said analyte.
Claim 51
Dependent Claim
30 words
The method of claim 39 wherein said first sbp member is analogous to said analyte and said second sbp member is complementary to said analyte and said first sbp member.
Claim 52
Dependent Claim
24 words
The method of claim 39 wherein said singlet oxygen is generated by irradiating said assay medium with light of wavelength of from 450-950 nm.
Claim 53
Independent Claim
152 words
A method for determining an analyte, said method comprising: combining in an assay medium either simultaneously or wholly or partially sequentially (1) a medium suspected of containing an analyte, (2) a label reagent comprising a first member of a specific binding pair (sbp member) bound to a compound capable of chemiluminescence upon reaction with singlet oxygen ("said compound"), (3) a singlet oxygen generator, and (4) a reagent comprising a fluorescent energy acceptor bound to, or capable of becoming bound to, a second sbp member, under conditions wherein an sbp member complex involving said label reagent and said energy acceptor is formed in relation to the presence of analyte in said medium and energy from said compound is capable of activating said fluorescent energy acceptor, activating said singlet oxygen generator, and examining said assay medium for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 54
Independent Claim
152 words
A method for determining an analyte, said method comprising: combining in an assay medium either simultaneously or wholly or partially sequentially (1) a medium suspected of containing an analyte, (2) a label reagent comprising a first member of a specific binding pair (sbp member) bound to a compound capable of chemiluminescence upon reaction with singlet oxygen ("said compound"), (3) a singlet oxygen generator, and (4) a reagent comprising a fluorescent energy acceptor bound to, or capable of becoming bound to, a second sbp member, under conditions wherein an sbp member complex involving said label reagent and said energy acceptor is formed in relation to the presence of analyte in said medium and energy from said compound is capable of activating said fluorescent energy acceptor, activating said singlet oxygen generator, and examining said assay medium for a signal, the presence or intensity thereof being related to the amount of analyte in said medium.
Claim 55
Dependent Claim
13 words
The method of claim 50 wherein said singlet oxygen generator is a dye.
Claim 56
Dependent Claim
21 words
The-method of claim 51 wherein said dye is selected from the group consisting of methylene blue, rose bengal, porphyrins, and phthalocyanines.
Claim 57
Dependent Claim
25 words
The method of claim 50 wherein said analyte and said sbp members are each independently selected from the group consisting of ligands, receptors, and polynucleotides.
Claim 58
Dependent Claim
26 words
The method of claim 50 wherein said chemiluminescent compound contains an olefin group and one or more electron donating substituents in conjugation with said olefin group.
Claim 59
Dependent Claim
23 words
The method of claim 50 wherein said compound is selected from the group consisting of 9-alkylidene acridans, enol ethers, 9-alkylidene xanthanes and enamines.
Claim 60
Dependent Claim
15 words
The method of claim 50 wherein said first sbp member is complementary to said analyte.
Claim 61
Dependent Claim
16 words
The method of claim 50 wherein said a second sbp member is complementary to said analyte.
Claim 62
Dependent Claim
29 words
The method of claim 50 wherein said first sbp is analogous to said analyte and said second sbp member is complementary to said analyte and said first sbp member.
Claim 63
Dependent Claim
25 words
The method of claim 50 wherein said singlet oxygen generator is activated by irradiating said assay medium with light of wavelength of from 450-950 nm.
Claim 64
Independent Claim
97 words
A method for determining a polynucleotide analyte, said method comprising: combining in an assay medium, (1) a sample suspected of containing a polynucleotide analyte and (2) a label reagent comprising a photochemically activatable chemiluminescent compound (PACC) bound to a polynucleotide, at least a portion of which is capable of hybridizing with said polynucleotide analyte, under conditions wherein said label reagent hybridizes with said polynucleotide analyte if present, irradiating said assay medium to activate said PACC, and measuring the fluorescent energy emitted from said medium, the amount thereof being related to the amount of analyte in said sample.
Claim 65
Independent Claim
97 words
A method for determining a polynucleotide analyte, said method comprising: combining in an assay medium, (1) a sample suspected of containing a polynucleotide analyte and (2) a label reagent comprising a photochemically activatable chemiluminescent compound (PACC) bound to a polynucleotide, at least a portion of which is capable of hybridizing with said polynucleotide analyte, under conditions wherein said label reagent hybridizes with said polynucleotide analyte if present, irradiating said assay medium to activate said PACC, and measuring the fluorescent energy emitted from said medium, the amount thereof being related to the amount of analyte in said sample.
Claim 66
Dependent Claim
20 words
The method of claim 60 wherein said assay medium also includes a fluorescent compound capable of activation by said PACC.
Claim 67
Dependent Claim
16 words
The method of claim 61 wherein said fluorescent compound is capable of binding to a polynucleotide.
Claim 68
Dependent Claim
40 words
The method of claim 61 wherein said fluorescent compound is bound to a polynucleotide, at least a portion of which is capable of hybridizing with said polynucleotide analyte at a region other than that with which said label reagent hybridizes.
Claim 69
Dependent Claim
20 words
The method of claim 61 wherein said fluorescent compound is selected from the group consisting of coumarins and xanthene dyes.
Claim 70
Dependent Claim
25 words
The method of claim 60 wherein said PACC contains an olefin group and one or more electron donating substituents in conjugation with said olefin group.
Claim 71
Dependent Claim
23 words
The method of claim 60 wherein said PACC is selected from the group consisting of 9-alkylidene acridans, enol ethers, 9-alkylidene xanthanes and enamines.
Claim 72
Dependent Claim
23 words
The method of claim 60 wherein said medium is irradiated with light having wavelengths between 450 and 950 nm to activate said PACC.
Claim 73
Dependent Claim
23 words
The method of claim 60 wherein said medium comprises a dye capable in the excited state of activating molecular oxygen to singlet oxygen.
Claim 74
Dependent Claim
20 words
The method of claim 68 wherein said medium is irradiated with light of wavelength 450-950 nm to excite said dye.
Claim 75
Dependent Claim
13 words
The method of claim 60 wherein said polynucleotide analyte is DNA or RNA.
Claim 76
Dependent Claim
13 words
The method of claim 61 wherein said polynucleotide analyte is DNA or RNA.
Claim 77
Dependent Claim
18 words
A composition comprising a photochemically activated chemiluminescent compound (PACC) associated with a member of a specific binding pair.
Claim 78
Dependent Claim
18 words
A composition comprising a photochemically activated chemiluminescent compound (PACC) associated with a member of a specific binding pair.
Claim 79
Dependent Claim
12 words
The composition of claim 72 wherein said PACC contains an olefin group.
Claim 80
Dependent Claim
25 words
The composition of claim 72 wherein said PACC contains an olefin group and one or more electron donating substitutents in conjugation with said olefin group.
Claim 81
Dependent Claim
22 words
The composition of claim 72 wherein said PACC is selected from the group consisting of 9-alkyline-N-alkyl acridans, enolethers, enamines, and 9-alkylidene xanthenes.
Claim 82
Dependent Claim
20 words
The composition of claim 72 wherein said sbp member is selected from the group consisting of receptors, ligands, and polynucleotides.
Claim 83
Dependent Claim
35 words
A kit comprising in packaged combination: (1) a composition comprising a photochemically activatable chemiluminescent compound (PACC), having bound thereto a specific binding pair (sbp) member, and (2) a photosensitizer which is not in said composition.
Claim 84
Dependent Claim
35 words
A kit comprising in packaged combination: (1) a composition comprising a photochemically activatable chemiluminescent compound (PACC), having bound thereto a specific binding pair (sbp) member, and (2) a photosensitizer which is not in said composition.
Claim 85
Dependent Claim
11 words
The kit of claim 77 wherein said photosensitizer is a dye.
Claim 86
Dependent Claim
22 words
The kit of claim 78 wherein said dye is selected from the group consisting of methylene blue, rose bengal, porphyrins, and phthalocyanines.
Claim 87
Dependent Claim
20 words
The kit of claim 77 wherein said sbp member is selected from the group consisting of receptors, ligands, and polynucleotides.
Claim 88
Dependent Claim
12 words
The kit of claim 77 wherein said PACC contains an olefin group.
Claim 89
Dependent Claim
25 words
The kit of claim 77 wherein said PACC contains an olefin group and one or more electro donating substitutents in conjugation with said olefin group.
Claim 90
Dependent Claim
22 words
The kit of claim 77 wherein said PACC is selected from the group consisting of 9-alkylidene-N-alkyl acridans, enolethers, 9-alkylidene xanthenes and enamines.
Claim 91
Dependent Claim
10 words
The kit of claim 83 which comprises an energy acceptor.
Claim 92
Dependent Claim
15 words
The kit of claim 84 wherein said energy acceptor is bound to an sbp member.
Claim 93
Dependent Claim
20 words
The kit of claim 85 where said sbp member is selected from the group consisting of receptors, ligands, and polynucleotides.
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