Patent ID: 6794148
Filing Date: 2004-09-21
Classification: G01N

Abstract:
A method of analyzing the relative binding affinities of at least a first ligand and a second ligand to a target using a tandem column chromatography system, the method comprising the steps of:(a) providing a solution comprising a plurality of mixed ligands and a target, wherein the plurality of mixed ligands comprises at least a first ligand and a second ligand; (b) introducing a sample of the solution to a first column, the first column comprising a size exclusion medium; (c) passing a number of column volumes of a solvent free of the target and ligands which bind to the target through the first column at a linear velocity; (d) introducing an early portion of the eluant from 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 sample of the solution to the first column; (f) passing a number of column volumes of a solvent free of the target and ligands which bind to the target through the first column at another linear velocity; (g) introducing an early portion of the eluant from 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 at a decreased linear fluid velocity correlates to a higher relative binding affinity for that ligand to the target.