title: Affinity surfactants as reversibly bound ligands for high-performance affinity chromatography.
abstract: Pyridine was coupled covalently to a nonionic ethoxylated alcohol: octaethylene glycol n-hexadecyl ether. This modified surfactant was found to be a reversible, competitive inhibitor of horse serum cholinesterase. The surfactant bound irreversibly, in aqueous media, to octadecyl-bounded reverse phase silica particles commonly used for high-performance liquid chromatography. The amount of ligand bound was found to be 550 mumol/ml of packing, a concentration that is over 100 times higher than what can be normally bound to agarose affinity chromatography supports. With this packing, a 280-fold purification of cholinesterase from horse serum and a 79-fold purification of human serum cholinesterase were accomplished, with yields greater than 80%, using a 2-cm-long column and a 7-min elution time. The affinity surfactant could be eluted from the column using a 6:4 (v/v) mixture of methanol and isopropanol. This technique should be generally applicable in the development of biospecific supports for high-performance affinity chromatography.
authors.affiliation: Department of Chemical Engineering, North Carolina State University, Raleigh 27695.
authors.forename: J L
authors.initials: JL
authors.lastname: Torres
authors.forename: R
authors.initials: R
authors.lastname: Guzman
authors.forename: R G
authors.initials: RG
authors.lastname: Carbonell
authors.forename: P K
authors.initials: PK
authors.lastname: Klipatrick
mesh_terms: D000818:Animals; D001665:Binding Sites; D002800:Cholinesterase Inhibitors / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D006736:Horses; D006801:Humans; D011726:Pyridinium Compounds; D013501:Surface-Active Agents / Q000494:pharmacology*
chemical_list: D002800:Cholinesterase Inhibitors; D011726:Pyridinium Compounds; D013501:Surface-Active Agents