Dry biochemical assay plate and method for making the same
First Claim
1. An assay plate for detecting the presence of a first predetermined type of oligonucleotide in a solution containing an unknown oligonucleotide, the assay plate comprising:
- a fused silica substrate; and
a dried aliquot of a first known oligonucleotide, the dried aliquot covalently bonded to the fused silica substrate, the known oligonucleotide operative to bind said predetermined type of oligonucleotide upon contact with a solution containing such predetermined type of oligonucleotide.
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Abstract
An assay plate for detecting the presence of a mobile reactant that binds to a immobilized reactant and the methods of making and using the same. An assay plate according to the present invention includes a substrate and at least one dried aliquot of the immobilized reactant, the immobilized reactant being bound to the surface of the substrate. The immobilized reactant binds the mobile reactant when a solution containing the mobile reactant is brought into contact with the immobilized reactant. The mobile and immobilized reactants may be any pair of biological compounds that have a specific affinity for one another. For example the reactants may be nucleic acids or antibody-antigen pairs. The preferred embodiment of an assay plate according to the present invention includes a plurality of assay spots, each spot having a different immobilized reactant or concentration thereof. The preferred method for fabricating an assay plate according to the present invention includes the steps of binding the immobilized reactant to the substrate, washing the substrate to remove any immobilized reactant that is not bound to the substrate and then drying the substrate. The dried assay plates are preferably stored in a water-proof container until used. An assay utilizing an assay plate according to the present invention is carried out by bringing a solution containing the mobile reactant into contact with the dried aliquot or aliquots on the assay plate. The assay plate is then washed to removed unbound material and the amount of mobile reactant bound to the washed assay plate determined. In the preferred embodiment of the present invention, the washed assay plate is dried prior to measuring the amount of mobile reactant bound to the washed assay plate.
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Citations
18 Claims
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1. An assay plate for detecting the presence of a first predetermined type of oligonucleotide in a solution containing an unknown oligonucleotide, the assay plate comprising:
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a fused silica substrate; and a dried aliquot of a first known oligonucleotide, the dried aliquot covalently bonded to the fused silica substrate, the known oligonucleotide operative to bind said predetermined type of oligonucleotide upon contact with a solution containing such predetermined type of oligonucleotide. - View Dependent Claims (2, 3, 4, 5)
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6. A method for making an assay plate for detecting the presence of a mobile oligonucleotide that binds to an immobilized known oligonucleotide, said method comprising the steps of:
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covalently binding said known oligonucleotide to a fused silica substrate to immobilize the known oligonucleotide; washing said substrate to remove any of said known oligonucleotide that fails to bind to said substrate; and drying said substrate and said bound immobilized oligonucleotide. - View Dependent Claims (7, 8, 9, 10)
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11. A method for detecting a mobile nucleic acid comprising the steps of:
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providing an assay plate of fused silica having a dried aliquot of an immobilized nucleic acid covalently bound thereon, said immobilized nucleic acid binding said mobile nucleic acid when both said immobilized nucleic acid and said mobile nucleic acid are in a wet state; bringing a solution containing said mobile nucleic acid into contact with said dried aliquot; washing said assay plate; treating with a dye that binds to one of said immobilized nucleic acid or said mobile nucleic acid; and determining the amount of mobile nucleic acid bound to said washed assay plate by measuring the dye. - View Dependent Claims (12, 13, 14, 15)
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16. A method for making an assay plate for detecting the presence of a mobile oligonucleotide that binds to an immobilized known oligonucleotide, said method comprising the steps of:
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covalently binding said known oligonucleotide to a fused silica substrate to immobilize the know oligonucleotide; washing said substrate to remove any of said known oligonucleotide that fails to bind to said substrate; and drying said substrate and said bound immobilized oligonucleotide, wherein the step of covalently binding the immobilized oligonucleotide to a fused silica substrate comprises; coating the substrate with a solution of amino propyl thriethoxy silane; linking the oligonucleotide that is to be immobilized to a linker; depositing the linked oligonucleotide to the coated substrate; and incubating the substrate, and wherein the step of coating the substrate comprises coating a surface of the substrate with a one percent solution of amino propyl triethoxy silane in ninety-five percent ethanol, and incubating at room temperature in a covered enclosure.
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17. A method for making an assay plate for detecting the presence of a mobile oligonucleotide that binds to an immobilized known oligonucleotide said method comprising the steps of:
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covalently binding said known oligonucleotide to a fused silica substrate to immobilize the known oligonucleotide; washing said substrate to remove any of said known oligonucleotide that fails to bind to said substrate; and drying said substrate and said bound immobilized oligonucleotide, wherein the step of covalently binding the immobilized oligonucleotide to a fused silica substrate comprises; coating the substrate with a solution of amino propyl thriethoxy silane; linking the oligonucleotide that is to be immobilized to a linker; depositing the linked oligonucleotide to the coated substrate; and incubating the substrate, and wherein the linker comprises Bis succinimydl suberate-homobifunctional NHS-ester.
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18. A method for making an assay plate for detecting the presence of a mobile oligonucleotide that binds to an immobilized known oligonucleotide, said method comprising the steps of:
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covalently binding said known oligonucleotide to a fused silica substrate to immobilize the know oligonucleotide; washing said substrate to remove any of said known oligonucleotide that fails to bind to said substrate; and drying said substrate and said bound immobilized oligonucleotide, wherein the drying step is carried out in an atmosphere of nitrogen.
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Specification