Easily regenerable affinity chromatographic zirconia-based support with concanavalin A as a model ligand.
The goal of this work was to develop a generic approach for producing affinity chromatographic columns which can be regenerated. Concanavalin A (Con A) was immobilized adsorptively by an in situ method onto a zirconium dioxide (zirconia) chromatographic support and used to resolve chromophorically labeled monosaccharides. The Con A was then removed from the zirconia by flushing with base. The same column was regenerated by applying a fresh aliquot of Con A. This cycle was repeated several times to demonstrate consistency in the loading capacity and the stability of the underlying zirconia support. Finally we used glutaraldehyde to cross-link the Con A to increase the long-term stability of the column. Hydrolyzing the protein with acid allowed it to be removed under alkaline conditions and the column regenerated simply by adding more Con A followed by glutaraldehyde cross-linking.
Glavanovich
M H
MH
Department of Chemistry, University of Minnesota, Minneapolis 55455-0431.
Carr
P W
PW
D000327:Adsorption; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A / Q000737:chemistry*; D003432:Cross-Linking Reagents; D005976:Glutaral / Q000737:chemistry; D006868:Hydrolysis; D009005:Monosaccharides / Q000302:isolation & purification*; D015040:Zirconium / Q000737:chemistry*
D003432:Cross-Linking Reagents; D009005:Monosaccharides; D003208:Concanavalin A; D015040:Zirconium; C028541:zirconium oxide; D005976:Glutaral