Concanavalin a attached poly(p-chloromethylstyrene) beads for glycoenzyme separation

摘要

Crosslinked poly(p-chloromethylstyrene) (PCMS) beads were produced by suspension polymerization. The beads had a nonporous but reasonably rough surface structure. Because of this property, a relatively high external surface area (i.e.,14.1 m(2)/g) was obtained with the proposed carrier. A biospecific ligand commonly used in the affinity chromatography of various glycoenzymes, concanavalin A (Con A), was covalently attached onto the bead surface by a direct chemical reaction. PCMS/Con A beads were then used for the separation of a model glycoenzyme, invertase, from its crude solution. The appropriate invertase adsorption and desorption conditions of the affinity system were investigated. The desorption of invertase from the carrier was achieved by the use of methyl-alpha-D-mannopyranoside (MMP) as the counterligand. The effects of the MMP and salt concentrations and the temperature on the desorption behavior of invertase were investigated. The MMP concentration and temperature were found to be the dominant parameters controlling the desorption. Iterative experiments involving five reversible adsorption-desorption cycles were performed with the same particles to monitor the changes that occurred in the invertase adsorption-desorption capacities of Con A immobilized beads. The reversible adsorption-desorption stability was significantly improved by the crosslinking of Con A immobilized on the beads. (C) 2004 Wiley Periodicals, Inc. [References: 61]