Methods for conducting specific binding assays
First Claim
1. A method for separating bound label from unbound label within an assay mixture formed within an assay vessel, wherein said assay mixture includes one or more binding components, label bound to at least some of said binding components, and a substantially aqueous solution containing unbound label, said binding components differing in apparent density from said aqueous solution, comprising:
- contacting a primary layer with an assay mixture formed within an assay vessel, said primary layer being immiscible with both said unbound label and said binding components, and of different density than said binding components; and
subjecting said assay mixture to conditions sufficient to cause said binding components and said unbound label to separate, based upon the comparative density of said binding components and the aqueous solution containing said unbound label.
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Abstract
Methods and devices for separating bound label from unbound label within an assay mixture and for predispensing assay reactants in self-contained assay vessels, as well as a method for detecting the presence and/or amount of an analyte within a fluid sample, and a reusable detection vessel for use therein and with specific binding assays in general are disclosed. Significant to the separation of bound label from unbound label is the use of a cushion comprising generally one primary layer and in some cases one or more secondary layers.
90 Citations
16 Claims
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1. A method for separating bound label from unbound label within an assay mixture formed within an assay vessel, wherein said assay mixture includes one or more binding components, label bound to at least some of said binding components, and a substantially aqueous solution containing unbound label, said binding components differing in apparent density from said aqueous solution, comprising:
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contacting a primary layer with an assay mixture formed within an assay vessel, said primary layer being immiscible with both said unbound label and said binding components, and of different density than said binding components; and subjecting said assay mixture to conditions sufficient to cause said binding components and said unbound label to separate, based upon the comparative density of said binding components and the aqueous solution containing said unbound label. - View Dependent Claims (3, 4, 5, 6, 7)
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2. The method of claim I wherein said primary layer is immiscible with water.
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8. A method for separating bound label from unbound label within an assay mixture, wherein said assay mixture includes one or more binding components, non-isotopic label bound to at least some of said binding components, and a substantially aqueous solution containing unbound label, said binding components differing in apparent density from said aqueous solution, comprising:
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contacting a primary layer with an assay mixture, said primary layer being immiscible with both said bound label and said binding components, and of differing density than said binding components; and subjecting said assay mixture to conditions sufficient to cause said binding components and said unbound label to separate, based upon the comparative density of said binding components and the aqueous solution containing said unbound label.
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9. A method for detecting the presence or amount of an analyte within a fluid sample, comprising:
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combining said fluid sample with a reaction mixture to form an assay mixture, said assay mixture formed within an assay vessel, wherein said assay mixture contains one or more binding components and label, at least some of said label and some of said analyte binding, directly or indirectly, to said binding components; contacting said assay mixture with a primary layer, said binding components and unbound label being immiscible with and of different density than said primary layer; subjecting said assay mixture to conditions sufficient to cause said binding components and said unbound label to separate, based upon the comparative density of said binding components and the assay mixture containing said unbound label; and detecting the label bound to said binding components and therefrom determining the presence or amount of said analyte. - View Dependent Claims (10, 11, 12, 13, 14)
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15. A method for detecting the presence or amount of an analyte within a fluid sample, comprising:
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combining said fluid sample with a reaction mixture to form an assay mixture, wherein said assay mixture contains one or more binding components and non-isotopic label, at least some of said label and some of said analyte binding, directly or indirectly, to said binding components; contacting said assay mixture with a primary layer, said binding components and unbound label being immiscible with and of different density than said primary layer; subjecting said assay mixture to conditions sufficient to cause said binding components and said unbound label to separate, based upon the comparative density of said binding components and the assay mixture containing said unbound label; and detecting the label bound to said binding components and therefrom determining the presence or amount of said analyte. - View Dependent Claims (16)
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Specification