Methods and kits for the amplification of thin film based assays
First Claim
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1. A thin film detection method for detecting an analyte of interest, comprising;
- providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers forming a thin film comprising an adhering attachment layer in contact with said substrate to which is affixed a primary receptive material which specifically binds said analyte of interest,reacting said device with a sample potentially comprising said analyte under conditions in which said analyte binds to said receptive material, andreacting bound analyte with a reagent which specifically binds said analyte and forms a thin film and becomes part of said thin film on the substrate of said device and creates a mass change of said thin film on said substrate,applying polarized light to the surface of said device, anddirectly measuring the change in mass of said thin film on the substrate of said device by reflection of polarized light, as an indication of the presence of said analyte in said sample.
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Abstract
Method for detecting an analyte of interest, comprising the steps of providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers comprising an adhering attachment layer to which is affixed a receptive material which specifically interacts with the analyte of interest; reacting the device with a sample potentially comprising the analyte under conditions in which the analyte binds to the receptive material; and reacting bound analyte with a reagent which creates a mass change on the surface of the device.
271 Citations
26 Claims
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1. A thin film detection method for detecting an analyte of interest, comprising;
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providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers forming a thin film comprising an adhering attachment layer in contact with said substrate to which is affixed a primary receptive material which specifically binds said analyte of interest, reacting said device with a sample potentially comprising said analyte under conditions in which said analyte binds to said receptive material, and reacting bound analyte with a reagent which specifically binds said analyte and forms a thin film and becomes part of said thin film on the substrate of said device and creates a mass change of said thin film on said substrate, applying polarized light to the surface of said device, and directly measuring the change in mass of said thin film on the substrate of said device by reflection of polarized light, as an indication of the presence of said analyte in said sample. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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2. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers forming a thin film comprising, an adhering attachment layer in contact with said upper layer of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a secondary receptive material specific for said analyte of interest conjugated to an enzyme such that said secondary receptive material conjugated to said enzyme specifically binds said analyte; c) contacting said device with said sample comprising said analyte bound to said secondary receptive material complexed with said enzyme such that said primary receptive material binds analyte bound to said secondary receptive material complexed with said enzyme; d) contacting said enzyme bound to said analyte via said secondary receptive material with an enzyme substrate and incubating for a period of time sufficient to cause precipitation of product from the interaction of said enzyme substrate and said enzyme such that said product becomes part of said thin film on said upper surface of said substrate of said device and creates a mass change of said thin film; e) applying polarized light to the surface of said device; and f) directly measuring said mass change of said thin film where said thin film includes said precipitation product relative to said thin film comprising said antibody layer to which analyte has not bound by reflection of polarized light, as an indication of the amount of said analyte in said test sample. - View Dependent Claims (25, 26)
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3. A thin film detection method for detecting an analyte of interest, comprising:
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providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers forming a thin film comprising an adhering attachment layer in contact with said substrate to which is affixed a primary receptive material which specifically binds said analyte of interest; reacting a sample potentially comprising said analyte with a reagent which specifically binds said analyte of interest; reacting said device with said sample under conditions in which said analyte bound to said reagent binds to said receptive material and forms a thin film on said thin film on the substrate of said device and creates a mass change of said thin film on the substrate greater than the change produced by the binding of the analyte alone; applying polarized light to the surface of said device; and directly measuring the change in mass of said thin film on the substrate of said device by reflection of polarized light, as an indication of the presence of said analyte in said sample.
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4. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers comprising, an adhering attachment layer in contact with said upper surface of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a reagent complexed to a secondary receptive material specific for said analyte of interest such that said secondary receptive material complexed to said reagent specifically binds said analyte; c) contacting said device with said sample so as to allow said analyte bound to said secondary receptive material complexed with said reagent to bind to said layer of antibodies on said substrate; d) contacting said reagent bound to analyte bound to said device with a precipitating agent and incubating for a period of time sufficient to cause precipitation of product from the interaction of said precipitating agent and said reagent such that said product becomes part of said thin film on the upper surface of said substrate of said device and creates a mass change of said thin film; e) applying polarized light to the surface of said device; and f) directly measuring said mass change of said thin film where said thin film includes said precipitation product relative to said thin film comprising said antibody layer to which analyte has not bound by reflection of polarized light, as an indication of the amount of said analyte in said test sample.
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5. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers forming a thin film comprising, an adhering attachment layer in contact with said upper surface of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a latex particle complexed to a secondary receptive material specific for said analyte of interest such that said secondary receptive material complexed with said latex particle specifically binds said analyte; c) contacting said device with said sample so as to allow said analyte bound to said secondary receptive material complexed with said latex particle to bind to said layer of antibodies on said substrate such that said latex particle bound to said analyte becomes part of said thin film on the upper surface of said substrate and results in a mass change of said thin film greater than change produced by binding of the analyte alone; d) applying polarized light to the surface of said device; and e) directly measuring mass change of said thin film where said thin film includes said latex particle relative to said thin film comprising said antibody layer to which analyte has not bound by reflection of polarized light, as an indication of the amount of said analyte in said test sample.
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6. A kit for an optical thin film assay for an analyte of interest comprising;
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a test device having an optically active surface comprising one or more layers forming a thin film comprising an adhering attachment layer in contact with said optically active surface to which is affixed a primary receptive material which specifically binds to said analyte, and a reagent which specifically reacts with said analyte bound to said surface so as to form a thin film and become part of said thin film on said optically active layer and result in an alteration of mass of said thin film of said optically active layer. - View Dependent Claims (7)
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8. A thin film detection method for detecting an analyte of interest, comprising;
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providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers forming a thin film comprising an adhering attachment layer in contact with said substrate to which is affixed a primary receptive material which specifically binds said analyte of interest, reacting said device with a sample potentially comprising said analyte under conditions in which said analyte binds to said receptive material, and reacting bound analyte with a reagent which specifically binds said analyte and forms a thin film and becomes part of said thin film on the substrate of said device thus creating a mass change of said thin film on the substrate, applying incident light to the surface of said device, and directly measuring the alteration of said incident light by an interference effect caused by the change in mass of said thin film on the substrate of said device as indication of the presence of said analyte in said sample.
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9. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers forming a thin film comprising, an adhering attachment layer in contact with said upper layer of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a secondary receptive material specific for said analyte of interest conjugated to an enzyme such that said secondary receptive material conjugated to said enzyme specifically binds said analyte; c) contacting said device with said sample comprising said analyte bound to said secondary receptive material complexed with said enzyme such that said primary receptive material binds analyte bound to said secondary receptive material complexed with said enzyme; d) contacting said enzyme bound to said analyte via said secondary receptive material with an enzyme substrate and incubating for a period of time sufficient to cause precipitation of product from the interaction of said enzyme substrate and said enzyme so that said product becomes part of said thin film on said upper surface of said substrate of said device and creates a mass change of said thin film; e) applying incident light to the surface of said device; and f) directly measuring the alteration of said incident light by an interference effect caused by said mass change of said thin film where said thin film includes said precipitation product relative to said thin film comprising said antibody layer to which analyte has not bound, as an indication of the amount of said analyte in said test sample.
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10. A thin film detection method for detecting an analyte of interest, comprising:
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providing a detection device comprising a light reflective or transmissive substrate supporting one or more layers forming a thin film comprising an adhering attachment layer in contact with said substrate to which is affixed a primary receptive material which specifically binds said analyte of interest; reacting a sample potentially comprising said analyte with a reagent which specifically binds said analyte of interest; reacting said device with said sample under conditions in which said analyte bound to said reagent binds to said receptive material and forms a thin film and becomes part of said thin film on the substrate of said device and creates a mass change of said thin film on the substrate greater than the change produced by the binding of the analyte alone; applying incident light to the surface of said device; and directly measuring the alteration of said incident light by an interference effect caused by the change in mass of said thin film on the substrate of said device as indication of the presence of said analyte in said sample.
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11. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers comprising, an adhering attachment layer in contact with said upper layer of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a reagent complexed to a secondary receptive material specific for said analyte of interest such that said secondary receptive material bound to said reagent specifically binds said analyte; c) contacting said device with said sample so as to allow said analyte bound to said secondary receptive material complexed with said reagent to bind to said layer of antibodies on said substrate; d) contacting said reagent bound to analyte bound to said device with a precipitating agent and incubating for a period of time sufficient to cause precipitation of product from the interaction of said precipitating agent and said reagent such that said product becomes part of said thin film on the upper surface of said substrate of said device and creates a mass change of said thin film; e) applying incident light to the surface of said device; and f) directly measuring the alteration of said incident light by an interference effect caused by said mass change of said thin film where said thin film includes said precipitation product relative to said thin film comprising said antibody layer to which analyte has not bound as an indication of the amount of said analyte in said test sample.
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12. A method for detecting an analyte of interest in a sample, comprising the steps of:
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a) providing a thin film optical immunoassay device comprising a substrate, having an upper and a lower surface, and supporting on its upper surface one or more layers forming a thin film comprising, an adhering attachment layer in contact with said upper surface of said substrate, and a primary receptive material comprising a layer of antibodies specific for said analyte of interest bound to said attachment layer; b) contacting said sample with a latex particle complexed to a secondary receptive material specific for said analyte of interest such that said secondary receptive material complexed with said latex particle specifically binds said analyte; c) contacting said device with said sample so as to allow said analyte bound to said secondary receptive material complexed with said latex particle to bind to said layer of antibodies on said substrate such that said latex particle becomes part of said thin film on the upper surface of said substrate and results in a mass change of said thin film greater than change produced by binding of the analyte alone; d) applying incident light to the surface of said device; and e) directly measuring the alteration of said incident light by an interference effect caused by the change in mass of said thin film on the substrate where said thin film includes said latex particle relative to said thin film comprising said antibody layer to which analyte has not bound, as an indication of the amount of said analyte in said test sample.
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Specification