Polyelectrolytic internal calibration system of a flow-through assay
First Claim
1. A method for detecting an analyte within a test sample comprising:
- contacting conjugated probes with a test sample, the conjugated probes comprising detectable probes conjugated with a specific binding member, the conjugated probes being in fluid communication with a porous membrane and the conjugated probes having a net charge prior to contact of the conjugated probes with the test sample, the porous membrane defining a detection zone that contains a capture reagent immobilized therein, at least a portion of the conjugated probes or complexes thereof binding to the capture reagent to generate a detection signal, the porous membrane defining a calibration zone that contains a first polyelectrolyte substantially non-diffusively immobilized therein, conjugated probes that have passed through the detection zone binding to the first polyelectrolyte to generate a calibration signal, the first polyelectrolyte and the conjugated probes binding via ionic interaction between the two; and
comparing the detection signal to the calibration signal to determine the relative amount of analyte present in the test sample.
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Abstract
A flow-through assay for detecting the quantity of an analyte residing in a test sample is provided. The flow-through assay contains a porous membrane that is in fluid communication with probe conjugates that contain a specific binding member and a detectable probe. The porous membrane also defines a detection zone and a calibration zone. The calibration zone contains a polyelectrolyte substantially non-diffusively immobilized on the porous membrane. The polyelectrolyte is capable of generating a detectable calibration signal that can be readily compared (visually, quantitatively, and the like) to a detection signal to determine the amount of analyte in the test sample.
134 Citations
20 Claims
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1. A method for detecting an analyte within a test sample comprising:
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contacting conjugated probes with a test sample, the conjugated probes comprising detectable probes conjugated with a specific binding member, the conjugated probes being in fluid communication with a porous membrane and the conjugated probes having a net charge prior to contact of the conjugated probes with the test sample, the porous membrane defining a detection zone that contains a capture reagent immobilized therein, at least a portion of the conjugated probes or complexes thereof binding to the capture reagent to generate a detection signal, the porous membrane defining a calibration zone that contains a first polyelectrolyte substantially non-diffusively immobilized therein, conjugated probes that have passed through the detection zone binding to the first polyelectrolyte to generate a calibration signal, the first polyelectrolyte and the conjugated probes binding via ionic interaction between the two; and comparing the detection signal to the calibration signal to determine the relative amount of analyte present in the test sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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Specification