Preparation of high-density Concanavalin A adsorbent and its use for rapid, high-yield purification of peroxidase from horseradish roots

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 d...

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Bibliographic Details
Published in:Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2004-04, Vol.803 (2), p.237-241
Main Authors: Franco Fraguas, Laura, Batista-Viera, Francisco, Carlsson, Jan
Format: Article
Language:English
Subjects:
Adsorption
Analysis
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Chromatography, Affinity
Concanavalin A
Concanavalin A - chemistry
Electrophoresis, Polyacrylamide Gel
Fundamental and applied biological sciences. Psychology
General pharmacology
Horseradish Peroxidase - isolation & purification
Medical sciences
Peroxidase
Pharmacology. Drug treatments
Plant Roots - chemistry
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Summary: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
ISSN:1570-0232
1873-376X
DOI:10.1016/j.jchromb.2003.12.023