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US 2014/0273035 A1Published application
Assay with increased dynamic range
AssigneeAbbott Laboratories
InventorsBarry L. DOWELL, Susan GAYDA, Qiaoqiao RUAN, Joseph P. SKINNER, Sergey Y. TETIN
Life of this patent
Estimating the term…
Abstract
Provided herein are assays and kits useful for avoiding “prozone phenomenon” or “hook effect” and which expand the range of accurately measurable analyte concentrations.
Claims41
A kit comprising:
- i) a first analyte-binding molecule comprising a first label;
- ii) a second analyte-binding molecule comprising a second label, wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule, and
- iii) a third analyte-binding molecule attached to a solid support, wherein the third analyte-binding molecule can bind to analyte concurrently with either the first analyte-binding molecule or the second analyte-binding molecule.
A method of expanding the dynamic range of an assay, comprising:
- a) contacting a test sample suspected of comprising an analyte with a first analyte-binding molecule comprising a first label, a second analyte-binding molecule comprising a second label and a third analyte-binding molecule attached to a solid support under conditions that allow binding of:
- (i) the first analyte-binding molecule and the third analyte-binding molecule and
- (ii) the second analyte-binding molecule and the third analyte-binding molecule to the analyte, wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule, wherein the first analyte-binding molecule and the second analyte-binding molecule do not concurrently bind to the analyte;
- b) measuring the signal intensities of the first label of the first analyte-binding molecule bound to the analyte and of the second label of the second analyte-binding molecule bound to the analyte;
- and
- c) determining the concentration of analyte by comparing the signal intensities of the first label and the second label.
3 more elements
A kit comprising:
- i) a first analyte-binding molecule attached to a first solid support;
- ii) a second analyte-binding molecule attached to a second solid support, wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule;
- and
- iii) a third analyte-binding molecule comprising a label, wherein the third analyte-binding molecule can bind to analyte concurrently with either the first analyte-binding molecule or the second analyte-binding molecule.
1 more element
33-41. (canceled)
A method of expanding the dynamic range of an assay, comprising:
- a) contacting a test sample suspected of comprising an analyte with a first analyte-binding molecule attached to a first solid support, a second analyte-binding molecule attached to a second solid support, and a third analyte-binding molecule comprising a label under conditions that allow binding of:
- (i) the third analyte-binding molecule to the first solid support via the analyte bound to the first analyte-binding molecule;
- and
- (ii) the third analyte-binding molecule to the second solid support via the analyte bound to the second analyte-binding molecule;
- and
- wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule, wherein the first analyte-binding molecule and the second analyte-binding molecule do not concurrently bind to the analyte;
- b) measuring the signal intensities from the label of the third analyte-binding molecule bound to the first solid support and to the second solid support;
- and
- c) determining the concentration of analyte by comparing the signal intensities from the label of the third analyte-binding molecule bound to the first solid support and to the second solid support.
6 more elements
44-64. (canceled)
A kit comprising:
- i) a first analyte-binding molecule attached to a first solid support;
- ii) a second analyte-binding molecule attached to a second solid support, wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule;
- and
- iii) a tracer comprising said analyte or fragment thereof attached to a label, wherein said tracer can compete with said analyte for binding to either the first analyte-binding molecule or the second analyte-binding molecule.
1 more element
67-73. (canceled)
A method of expanding the dynamic range of an assay, comprising:
- a) contacting a test sample suspected of comprising an analyte with tracer comprising said analyte or fragment thereof attached to a label, a first analyte-binding molecule attached to a first solid support, a second analyte-binding molecule attached to a second solid support, wherein the binding affinity for the analyte of the first analyte-binding molecule is greater than that of the second analyte-binding molecule, wherein the first analyte-binding molecule and the second analyte-binding molecule do not concurrently bind to the analyte;
- b) measuring the signal intensities from the tracer bound to the first analyte-binding molecule on the first solid support and the second analyte-binding molecule on the second solid support;
- and
- c) establishing a flag value at or near the leveling off value (plateau) of the signal intensity of the tracer bound to the first analyte-binding protein attached to the first solid support.
1 more element
Dependencies are inferred from the claim text.