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Abstract

A method of analyzing the concentration of soluble analyte in a sample involves performing a competition assay using a predetermined amount of formed bodies to which are attached at least one analyte, varying known concentrations of an unlabeled ligand that binds to analyte, and a known concentration of ligand labeled with a detectable marker. After analysis in a flow cytometer, the test sample and a plurality of control samples generate data on curves formed by plotting signal vs. concentration of labeled ligand in controls (first), and vs. concentration of total labeled and unlabeled ligand in test samples (second). The concentration of unlabeled ligand that bound to soluble analytes in test samples is determined by evaluating the difference between the concentration that corresponds with the intersection of these two curves and the constant labeled ligand concentration in the test samples. The concentration of said soluble analytes in said sample is the product of that difference and the binding stoichiometry of the ligand to soluble analytes.

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First Claim