Lateral flow assay devices and methods for conducting assays

A method of detecting the quantity of an analyte present in a test solution, the method comprising:

• (a) providing a membrane with a test solution containing analyte;
• (b) providing a plurality of probes and probe conjugates upon the membrane;
• (c) binding the probe conjugates with the analyte to form probe analyte conjugates;
• (d) wherein the membrane comprises a first end and a second end, the membrane having a first capture reagent immobilized upon a detection zone, the membrane being configured for mobilizing a test solution containing probe-analyte conjugates from the first end to the second end of the membrane;
• (e) capturing within a detection zone probe-analyte conjugates, thereby forming immobilized signal generating sandwich complexes within the detection zone;
• (f) providing a calibration zone upon the membrane, the calibration zone comprising at least first and second control lines having predetermined amounts of a second capture reagent immobilized upon said control lines;
• (g) capturing, with the second capture reagent, probes upon the first and second control lines by forming control probe complexes upon said control lines;
• (h) generating a first set of control signals from the control probe complexes;
• (i) generating a second set of measured signals from the sandwich complexes in the detection zone;
• and
• (j) comparing the control signals to the measured signals to determine the quantity of analyte present in the test solution.