Capillary electrophoretic methods to detect new biologically active compounds in complex biological material

Abstract

This invention relates to a competitive-binding, capillary electrophoretic method of detecting new therapeutic regulatory (modulating) and diagnostic compounds in natural samples and other complex biological materials. The present method generally comprises mixing a preselected, detectable target with a sample of complex biological material to produce a first, sample/target mixture capillary electrophoresis apparatus. Subsequently, the first mixture is mixed with a pre-selected, tight-binding competitive ligand (TBCL), prior to produce a second, sample/target/TBCL mixture, for a predetermined optional incubation period sufficient to allow the TBCL to bind a pre-selected percentage of the available target in the absence of any other ligand. An aliquot of the second mixture is subsequently subjected to pre-optimized capillary electrophoresis, during which the migration of the target is monitored. The presence of a potential new compound is indicated by the increase in the peak area of the unbound target peak and/or decrease in the peak area of the TBCL/target complex peak. A capillary electrophoretic profile of the second mixture is produced, which may be compared to a reference standard. The reference standard typically comprises a capillary electrophoretic profile or migration pattern of the target when mixed with a TBCL in an absence of any other competing ligand under similar, pre-selected capillary electrophoretic conditions.