Apparatus for determining an analyte and method therefor
First Claim
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1. An apparatus for determining at least one analyte in a fluid sample, comprising:
- (a) a first porous zone comprising an immobilized, non diffusible reagent capable of specifically binding to at least one diffusible labeled reactant comprising a label conjugated to either said analyte, an analogue of said analyte or a specific binding partner for said analyte, and(b) a second porous zone comprising (i) a diffusible interactive reagent capable of reacting with the label portion of said diffusible labeled reactant to produce a diffusible detectable moiety and (ii) a means for separating said diffusible interactive reagent for said diffusible detectable moiety.
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Abstract
An apparatus for determining an analyte in a sample, as well as a method for carrying this out, are disclosed. The apparatus utilizes a first zone containing a labeled substance, and a second zone which permits separation of a mobile detectable moiety from unreacted reaction component. Detectable moiety is the product of reaction between reaction component and labeled substance.
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Citations
45 Claims
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1. An apparatus for determining at least one analyte in a fluid sample, comprising:
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(a) a first porous zone comprising an immobilized, non diffusible reagent capable of specifically binding to at least one diffusible labeled reactant comprising a label conjugated to either said analyte, an analogue of said analyte or a specific binding partner for said analyte, and (b) a second porous zone comprising (i) a diffusible interactive reagent capable of reacting with the label portion of said diffusible labeled reactant to produce a diffusible detectable moiety and (ii) a means for separating said diffusible interactive reagent for said diffusible detectable moiety. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. An apparatus for determining at least one analyte in a fluid sample, comprising:
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(a) a first zone comprising an immobilized, non-diffusible reagent capable of specifically binding to at least one diffusible reactant comprising analyte, labeled analyte, labeled analyte analogue or labeled analyte reaction partner, (b) a second zone comprising (i) a diffusible interactive reagent capable of reacting with the label of said diffusible detectable moiety and (ii) a means for separating said diffusible interactive reagent from said diffusible detectable moiety, and (c) at least one flow regulating means which regulates movement of a fluid sample introduced to said apparatus, wherein said flow regulating means has a flow rate different from the flow rate of at least a part of one of said first and second zones. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41)
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42. Method for determining at least one analyte in a fluid sample, comprising:
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(a) contacting said sample with a labeled binding partner which specifically binds to said analyte to form a mixture comprising (i) complexes of analyte and labeled binding partner and (ii) uncomplexed labeled binding partner, (b) contacting said mixture to an immobilized reagent which binds to uncomplexed labeled binding partner but not complexes of analyte and labeled binding partner, so as to remove uncomplexed labeled binding partner from said mixture, (c) contacting said mixture to an apparatus comprising a diffusible, interactive reagent capable of reacting with the label portion of said labeled binding partner to produce a detectable moiety in said mixture, (d) contacting said mixture with a means for separating detectable moiety from any unreacted diffusible interactive reagent, and (e) measuring one of unreacted diffusible interactive reagent, removed uncomplexed labeled binding partner or detectable moiety as a measure of analyte in said sample.
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43. Method for determining at least one analyte in a fluid sample, comprising:
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(a) contacting said sample with a labeled reactant comprising a label conjugated to either said analyte or an analogue of said analyte, to form a mixture; (b) contacting said mixture with an immobilized binding partner capable of binding to said analyte and to said labeled reagent to form complexes of (i) immobilized binding partner and analyte, and of (ii) immobilized binding partner and labeled reagent, (c) contacting said complex containing mixture with an interactive reagent capable of reacting with the label portion of said labeled reactant to form a detectable moiety, (d) contacting said complex containing mixture with means for separating unreacted interactive reagent and detectable moiety from each other, and (e) measuring one of unreacted interactive reagent, immobilized labeled reagent or detectable moiety as a measure of analyte in said sample.
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44. Method for determining at least one analyte in a fluid sample, comprising:
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(a) contacting said sample with a labeled reactant comprising a label conjugated to either said analyte or an analogue of said analyte, to form a mixture; (b) contacting said mixture with an immobilized binding partner capable of binding to said analyte and to said labeled reagent, to form complexes of immobilized binding partner and analyte, and of immobilized binding partner and labeled reagent; (c) separating said complexes from said mixture; (d) contacting complex free mixture with an interactive reagent capable of reacting with the label portion of said labeled reactant to produce a detectable moiety; (e) contacting said complex free mixture with a means for separating unreacted interactive reagent from said detectable moiety, and (f) measuring one of label in said separated complex detectable moiety or unreacted interactive reagent as a measure of analyte in said sample.
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45. Method for determining at least one analyte in a fluid sample, comprising:
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(a) contacting said sample with a complex of labeled reactant capable of specifically binding to said analyte and an immobilized binding partner for said labeled reactant, so as to form a mixture of displaced labeled reactant complexed to said analyte and free labeled reactant; (b) contacting said mixture to an immobilized reagent capable of specifically binding to free labeled reactant but not to complexes of labeled reactant and analyte, so as to remove free labeled reactant therefrom; (c) contacting said mixture to an interactive reagent which reacts with the label of said labeled reactant to form a detectable moiety; (d) contacting said mixture with a separation means for separating interactive reagent from detectable moiety; and (e) measuring immobilized labeled reactant, unreacted interactive reagent or detectable moiety as a measure of analyte in said sample.
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Specification