Avidin column as a highly efficient and stable alternative for immobilization of ligands for affinity chromatography.
The avidin/biotin system was applied as a general mediator in the adsorption/desorption or immobilization of biologically active macromolecules to solid supports. In this context, model biotinylated proteins (lectins and antibodies) were attached to avidin-coupled Sepharose. As examples for affinity chromatography, peanut agglutinin and anti-transferrin antibody were used to isolate asialofetuin and transferrin, respectively. The capacity and product yields were significantly better than those achieved with conventional affinity chromatography on CNBr-activated Sepharose columns containing the same lectin or antibody. Moreover, the columns were characterized by improved stability properties exhibiting remarkably low levels of leakage.
Bayer
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EA
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Wilchek
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M
D000906:Antibodies; D001360:Avidin; D001710:Biotin; D002846:Chromatography, Affinity / Q000379:methods*; D037102:Lectins; D008024:Ligands; D012685:Sepharose
D000906:Antibodies; D037102:Lectins; D008024:Ligands; D001360:Avidin; D001710:Biotin; D012685:Sepharose