Toroidal Coil Countercurrent Chromatography in the Affinity Partitioning of Nicotinic Cholinergic Receptor Enriched Membranes

Abstract

A variation on the aqueous polymer phase partition method, affinity partitioning, has proved suitable for the preparative scale purification of binding site enriched membrane fragments. The full resolving potential of the affinity partitioning technique often requires the utilization of multiple extraction procedures such as countercurrent distribution. In this report, we evaluate the combination of a newly developed countercurrent purification technique, toroidal coil chromatography, with affinity partitioning. This approach provides an efficient method for purification and characterization of membrane bound nicotinic cholinergic receptors. The relative merits of the toroidal coil chromatography technology and the more conventional thin-layer countercurrent distribution techniques are compared.