Reduction of the hook effect in membrane-based assay devices
First Claim
1. A flow-through assay device for detecting the presence or quantity of an analyte residing in a test sample, said flow-through assay device comprising a porous membrane, said porous membrane being in communication with conjugated detection probes capable of generating a detection signal, said porous membrane defining:
- a chromatographic zone within which a plurality of microporous particles are immobilized; and
a detection zone located downstream from said chromatographic zone, wherein a capture reagent is immobilized within said detection zone that is configured to bind to said conjugated detection probes, wherein said conjugated detection probes are capable of generating a detection signal while within said detection zone, wherein the amount of the analyte within the test sample is determined from said detection signal.
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Abstract
A membrane-based assay device for detecting the presence or quantity of an analyte residing in a test sample is provided. The device utilizes a chromatographic zone on which is disposed a plurality of microporous particles. The chromatographic zone can effectively reduce the “hook effect” in a simple, efficient, and relatively inexpensive manner. In particular, the plurality of microporous particles allows larger-sized analyte/probe complexes to reach the detection zone before the uncomplexed analyte. Because the uncomplexed analyte is substantially inhibited from competing with the complexes for the binding sites at the detection zone, the incidence of “false negatives” may be limited, even at relatively high analyte concentrations.
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Citations
29 Claims
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1. A flow-through assay device for detecting the presence or quantity of an analyte residing in a test sample, said flow-through assay device comprising a porous membrane, said porous membrane being in communication with conjugated detection probes capable of generating a detection signal, said porous membrane defining:
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a chromatographic zone within which a plurality of microporous particles are immobilized; and a detection zone located downstream from said chromatographic zone, wherein a capture reagent is immobilized within said detection zone that is configured to bind to said conjugated detection probes, wherein said conjugated detection probes are capable of generating a detection signal while within said detection zone, wherein the amount of the analyte within the test sample is determined from said detection signal. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 23, 24, 25, 26, 27, 28, 29)
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14. A flow-through, sandwich assay device for detecting the presence or quantity of an analyte residing in a test sample, said assay device comprising a porous membrane, said porous membrane being in communication with conjugated detection probes capable of generating a detection signal, said conjugated detection probes being configured to combine with the analyte in the test sample when contacted therewith such that analyte/probe complexes and uncomplexed analyte are formed, said porous membrane defining:
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a chromatographic zone within which a plurality of microporous particles are immobilized, said microporous particles being configured so that said uncomplexed analyte flows through said chromatographic zone at a slower rate than said analyte/probe complexes; and a detection zone located downstream from said chromatographic zone, wherein a capture reagent is immobilized within said detection zone that is configured to bind to said analyte/probe complexes so that said complexes generate a detection signal while within said detection zone, wherein the amount of the analyte within the test sample is determined from said detection signal. - View Dependent Claims (15, 16, 17, 18, 19, 20, 21, 22)
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Specification