Patent ID: 6358692
Filing Date: 2002-03-19
Classification: G01N

Abstract:
A method of analyzing the relative binding affinities of at least a first ligand and a second ligand to an immobilized target using a tandem column chromatography system, the method comprising the steps of:(a) providing a solution comprising a plurality of mixed ligands; (b) introducing an aliquot of the solution to a first column, the first column comprising a matrix having immobilized thereon a target which binds at least a first ligand and a second ligand of the plurality of mixed ligands; (c) introducing a number of column volumes of a solvent free of ligands which bind to the target into the first column, wherein the number of column volumes of solvent exit the first column; (d) eluting, after step (c), the first column directly into a second column to determine the presence or absence of the first ligand and the second ligand; (e) introducing another aliquot of the solution to the first column; (f) introducing another number of column volumes of a solvent free of ligands which bind to the target into the first column, wherein the other number of column volumes of solvent exit the first column; (g) eluting, after step (f), the first column directly into the second column to determine the presence or absence of the first ligand and the second ligand; and (h) determining the relative binding affinities of at least the first ligand and the second ligand for the target based on the results of steps (d) and (g), where the presence of one of the first ligand and the second ligand after an increased number of column volumes of solvent are introduced into the first column correlates to a higher relative binding affinity for that ligand.