Comparison of modified supports on the base of glycoprotein interaction studies and of adsorption investigations

• Published in: Journal of Chromatography A Vol. 1092; no. 1; pp. 76 - 88
• Main Authors: Rosenfeld, Henning, Aniulyte, Jolita, Helmholz, Heike, Liesiene, Jolanta, Thiesen, Peter, Niemeyer, Bernd, Prange, Andreas
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 21.10.2005; Elsevier
• Subjects: Adsorption; Affinity separation; Analytical chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Cellulose; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography, Affinity; ConA; Electrophoresis, Polyacrylamide Gel; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Glycoprotein purification; Glycoproteins - chemistry; Glycoproteins - metabolism; Kinetics; Lectin; Lectins - chemistry; Ligands; Models, Molecular; Other chromatographic methods; Protein Binding; Proteins; Selective adsorbents; Silica; Supports; Surface modification; WGA; WGA; Surface modification; Lectin; Glycoprotein purification; Supports; Cellulose; Affinity separation; Selective adsorbents; Silica; ConA; Glucose oxidase; Multicomponent mixture; Adsorption isotherm; Immobilization; Affinity adsorbent; Separation method; Binding protein; Binding capacity; Affinity chromatography; Silica gel; Stationary phase; Purification; Enzyme; Glycoprotein; Concanavalin A; Protein; Surface treatment; Adsorption; Agglutinin; Cellulose derivatives; Isolation; Oxidoreductases; Kinetics; Fetuin; Comparative study
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2005.07.027

The features of matrices, suitable for affinity chromatography, have been extensively investigated and got subject for several reviews. But these investigations show, that there is still a lack in adsorbent characterization and a demand of comparative investigations of adsorbents, based on different materials with a range of different surface functionalities. In this work the performance of self-prepared silica and cellulose-based adsorbents were compared with commercially available polymeric supports. A model system was chosen comprising the lectins concanavalin A (ConA) and wheat germ agglutinin (WGA), which were covalently attached to the support matrices, combining the selectivity of the lectin–sugar interaction with the chemical and mechanical properties of the support that influence the efficacy of the prepared adsorbent. The verification of the different supports provides information about tayloring carbohydrate specific lectin adsorbents. The characterization outlines the main features of the different adsorbents and takes into account the properties of the pure supports. It encompasses immobilization kinetics and isotherms as well as the description of the binding capacity of the adsorbents by depicting adsorption isotherms. The separation performances were also investigated in terms of glycoprotein purification factors and recoveries. Further, detailed information about binding of GOD to immobilized ConA are obtained.