Comparison of Modified Supports on the Base of Glycoprotein Interaction Studies and of Adsorption Investigations
AbstractThe features of matrices, suitable for affinity chromatography, have been extensively investigated and got subject of several reviews. But all this investigations show, that there is still a lack in the methodology of the adsorbents 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 taylored silica based adsorbents of the Helmut-Schmidt University and cellulose-based supports of the Kaunas University of Technology were compared with commercially available 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 reaction with the chemical and mechanical properties of the support that determine the efficacy of the isolation procedure. The verification of the different supports should provide information for tayloring specific lectin adsorbents for carbohydrate recognition. The characterization outlines the main features of different adsorbents, takes into account the properties of the supports and encompasses the immobilization kinetics and isotherms as well as the description of the binding capacity of the adsorbent by depicting adsorption isotherms. The performances of the adsorbents were also investigated in terms of glycoprotein purification factors and recoveries.Further, detailed information about the lectin glycoprotein binding are obtained.