REDUCTION OF THE HOOK EFFECT IN MEMBRANE-BASED ASSAY DEVICES
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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.
99 Citations
40 Claims
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1-22. -22. (canceled)
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23. A method for detecting the presence or quantity of an analyte residing in a test sample, said method comprising:
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i) providing a 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;
ii) contacting a test sample containing the analyte with said conjugated detection probes so that analyte/probe complexes and uncomplexed analyte are formed; and
iii) allowing said analyte/probe complexes and said uncomplexed analyte to reach said chromatographic zone and then said detection zone, wherein said analyte/probe complexes reach said detection zone before said uncomplexed analyte. - View Dependent Claims (24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40)
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Specification