Amplified signal in binding assays
First Claim
1. A method for determining the presence or concentration of an analyte in a medium, said method comprising:
- providing a reaction mixture comprising in combination;
a medium suspected of containing an analyte;
a first specific binding pair member bound to a water-insoluble solid support;
a second specific binding pair member bound to a sensitizer, said sensitizer capable in its excited state of generating a reactive oxygen species, wherein the proximity of the first specific binding pair member with the second specific binding pair member is modulated by the presence of the analyte; and
digoxigenin-linked biotin linked to the solid support through a reactive oxygen cleavable linker;
incubating the reaction mixture;
exciting the sensitizer, said excitation of the sensitizer causing the formation of reactive oxygen, which cleaves the cleavable linker and releases digoxigenin-linked biotin from the solid support; and
detecting the released digoxigenin-linked biotin, the amount thereof being related to the amount of analyte in said medium.
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Abstract
Methods are disclosed for determining minute quantities of an analyte in a medium suspected of containing the analyte. One method comprises treating a medium suspected of containing an analyte under conditions such that the analyte, if present, causes a substrate having an oxidant cleavable linker and a photosensitizer to come into close proximity. The photosensitizer generates singlet oxygen which oxidatively cleaves the linker to form a product which can be detected in a sandwich detection assay such as LOCI. The amount of product detected is related to the amount of analyte in the medium. Compositions, kits, and compounds are also disclosed.
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Citations
3 Claims
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1. A method for determining the presence or concentration of an analyte in a medium, said method comprising:
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providing a reaction mixture comprising in combination; a medium suspected of containing an analyte; a first specific binding pair member bound to a water-insoluble solid support; a second specific binding pair member bound to a sensitizer, said sensitizer capable in its excited state of generating a reactive oxygen species, wherein the proximity of the first specific binding pair member with the second specific binding pair member is modulated by the presence of the analyte; and digoxigenin-linked biotin linked to the solid support through a reactive oxygen cleavable linker; incubating the reaction mixture; exciting the sensitizer, said excitation of the sensitizer causing the formation of reactive oxygen, which cleaves the cleavable linker and releases digoxigenin-linked biotin from the solid support; and detecting the released digoxigenin-linked biotin, the amount thereof being related to the amount of analyte in said medium. - View Dependent Claims (2, 3)
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Specification