title: Preparation of high-density Concanavalin A adsorbent and its use for rapid, high-yield purification of peroxidase from horseradish roots.
abstract: Preparation of Concanavalin A-adsorbents by immobilization on Sepharose activated with 1-cyano-4-(dimethylamino)-pyridinium tetrafluoroborate (CDAP-reagent) is reported. High immobilization yields of lectin (above 90%) were attained using an optimized CDAP-activating protocol. The effect of ligand density on the performance of the adsorbent for specific binding of glycoproteins was studied using horseradish peroxidase (HRP) as a model. Adsorption yields of pure HRP exceeding 90% were obtained with Con A-derivatives containing not < 20 mg of immobilized Con A/ml of packed gel. With lectin content of 2 mg/(ml of packed gel), only 20% of HRP was adsorbed. Purification of peroxidase from horseradish roots extract was successfully accomplished on Con A-Sepharose with high Con A content.
authors.affiliation: Cátedra de Bioquímica, Facultad de Química, CC 1157, Gral Flores 2124, Montevideo, Uruguay. lfranco@fq.edu.uy
authors.forename: Laura
authors.initials: L
authors.lastname: Franco Fraguas
authors.forename: Francisco
authors.initials: F
authors.lastname: Batista-Viera
authors.forename: Jan
authors.initials: J
authors.lastname: Carlsson
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006735:Horseradish Peroxidase / Q000302:isolation & purification*; D018517:Plant Roots / Q000737:chemistry*
chemical_list: D003208:Concanavalin A; D006735:Horseradish Peroxidase