EP0324616B1
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Template-directed photoligation
Assignee: BP Corp North America Inc
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Filing: 1989-01-12
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Granted: 1995-03-29
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Expires: 2009-01-12
Abstract
Abstract Methods, apparatus and compositions are presented for ligating ligands together which bind to a common target. One embodiment includes polynucleotide probes having photoreactive functional groups. The probes are capable of assuming substantially contiguous reactive positions on a target polynucleotide placing the photoreactive groups in juxtaposition. Activation of the photoreactive functional groups with radiant energy form a probe reaction product in which the probes are bound to each other.
Inventors
- 👤 Garfield Paul Royer
- 👤 Kenneth Alexander Cruickshank
- 👤 Larry Edward Morrison
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-31 00:51
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C07; C07D; C07D493/00; C07D493/02; C07D493/04; C07D311/00; C07D311/02; C07D311/04; C07D311/06; C07D311/08; C07D311/14; C07D311/16; C07D311/18; C12Q1/6813; C12Q1/6816; C12Q1/6827
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
127 words
A method for ligating nucleotide ligands which are capable of binding to substantially contiguous positions of a target molecule comprising the steps of: a) contacting the target with a first polynucleotide ligand and a second polynucleotide ligand wherein said first ligand includes a photoreactive functional group and said second ligand includes a photoreactive functional group wherein said photoreactive functional groups are capable of interacting to form a covalent bond between said first and second ligands upon activation, and said first and second ligand are capable of simultaneously binding to the target to form a target-first ligand-second ligand complex; and b) activating the reactive functional groups to form a covalent bond between the first and second ligands while the first and second ligands are bound to the target.
Claim 2
Independent Claim
148 words
A method for assaying a sample for target polynucleotide molecules comprising: a) contacting the sample under binding conditions with a first polynucleotide probe and a second polynucleotide probe wherein the first probe includes a photoreactive functional group and the second probe includes a photoreactive functional group, wherein said photoreactive functional groups are capable of interacting to form a covalent bond between the first and second probes upon activation when the first probe and second probe are placed in a reactive position to form a probe reaction product, and wherein the first probe and the second probe are capable of binding to the target molecule to form a target-first ligand-second ligand complex; b) subjecting the sample in the presence of the first probe and the second probe to radiant energy to form the probe reaction complex; and c) monitoring the sample for the presence of the probe reaction product.
Claim 3
Independent Claim
30 words
The method of either of Claims 1 or 2 wherein said photoreactive functional group has an extinction coefficient greater than 1,000 mole ⁻¹·1·cm⁻¹ to permit excitation to reactive energy levels.
Claim 4
Independent Claim
52 words
The method of either of Claims 1 or 2 wherein said photoreactive functional group is selected from the group of photoreactive functional groups consisting of coumarins, psoralens, anthracenes, pyrenes, carotenes, tropones, chromones, quinones, maleic anhydride, alkyl maleimide, olefins, ketones, azides, polyolefins characterized by conjugated double bonds and ketone functionality and derivatives thereof.
Claim 5
Dependent Claim
14 words
The method of Claim 4 wherein said photoreactive functional group is a coumarin derivative.
Claim 6
Dependent Claim
76 words
The method of Claim 5 wherein said coumarin is characterized by the following features: wherein, R groups 1 through 6 are selected from the group consisting of -H, -CH₃, -CH₂Cl, -CH₂Br, -CH₂I, -CH₂F, -CHCl₂, -CHBr₂, -CHI₂, -CHF₂, -CH₂OH, -CH₂OCH₃, -CH₂NH₂, -N₃, -COOH, -COOCH₃, -COOCH₂CH₃, -NH₂, -NO₂, -CBr₃, -CI₃, -CF₃, -CCl₃, -CH(CH₃)₂, -C(CH₃)₃, -Cl, -Br, -I, -F, and -O(CH₂) n CH₃(n=0 to 10) wherein one of the R groups includes a functionality for linking to a polynucleotide.
Claim 7
Dependent Claim
77 words
The method of claim 4 wherein said psoralen is characterized by the following features: wherein, R groups 1 through to 6 are selected from the group consisting of -H, -CH₃, -CH₂Cl, -CH₂Br, -CH₂I, -CH₂F, -CHCl₂, -CHBr₂, -CHI₂, -CHF₂, -CH₂OH, -CH₂OCH₃, -CH₂NH₂, -N₃, -COOH, -COOCH₃, -COOCH₂CH₃, -NH₂, -NO₂, -CBr₃, -CI₃, -CF₃, -CCl₃, -CH(CH₃)₂, -C(CH₃)₃, -Cl, -Br, -I, -F, and -O(CH₂) n CH₃(n=0 to 10) wherein one of the R groups includes a functionality for linking to a polynucleotide.
Claim 8
Dependent Claim
42 words
The method of Claim 2 wherein said first probe and said second probe are polynucleotide strands and said photoreactive functional group of said first probe and said photoreactive molecule of said second probe are located at the 3' and 5' termini respectively.
Claim 9
Dependent Claim
29 words
The method of Claim 2 wherein said photoreactive functional group undergoes photochemical reactions upon excitation with photons having a wavelength of approximately between 300 nm and 380 nm inclusive.
Claim 10
Dependent Claim
39 words
The method of Claim 2 wherein said photoreactive functional group is selected from the group consisting of coumarin-7-oxybutyric acid N-hydroxysuccinimide ester, 4-methylcoumarin-7-oxybutyric acid N-hyroxysuccinimide ester, 7-methoxycoumarin-6-oxybutyric acid N-hydroxysuccinimide ester, coumarin-5-oxybutyric acid N-hydroxysuccinimide ester, 6-methoxycoumarin-7-oxybutyric acid N-hydroxysuccinimide ester, and coumarin-6-isothiocyanate.
Claim 11
Dependent Claim
142 words
A kit for performing photoligation of substantially contiguous ligands comprising a reagent contacting a sample potentially containing a target molecule of interest, wherein the reagent includes a first probe and a second probe and the first probe and the second probe are polynucleotide strands capable of binding to the target molecule and wherein the first probe and the second probe have at least one photoreactive functional group capable of covalently binding the first probe and the second probe to form a probe reaction product in the presence of radiant energy when the first probe and second probe are placed in a reactive position, and the first probe and second probe are capable of binding to the target molecule in a reactive position and the reaction product has detectable features which are distinguishable from either the first probe or the second probe individually.
References Cited (Backward Citations)
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