Purification by Affinity Chromatography of Red Cell Membrane Acetylcholinesterase

Abstract

Over the past the methods for purifying integral membrane proteins have advanced considerably. In particular the onset of affinity chromatography (Jakoby and Wilchek 1974) has brought new incentives to this particular class of proteins. In order to be deployed successfully the following criteria have to be met: (A) A high affinity ligand covalently linked to an insoluble support via a spacer arm must warrant biospecific adsorption. After removal of all undesired protein, desorption should be carried out using similar biospecificity. (B) The amphiphilic membrane protein must be kept in solution throughout the purification procedure by the use of a suitable detergent. Alternatively membrane proteins may be solubilized by proteolytic enzymes, chaotropic ions or extraction with organic solvents.