Investigations on protein adsorption to agarose-dextran composite media.
A new stationary phase for protein purification was investigated with regard to its performance during capture of selected model proteins. The commercially available matrix consists of a porous agarose backbone, to which dextran is covalently attached. The dextran carries ion-exchange ligands, thus providing a binding space of high ligand density. Breakthrough of various proteins during frontal application to packed beds was measured and the experiments were analyzed in terms of equilibrium and breakthrough capacity. A significant increase of static capacity, as compared with conventional porous matrices, was found. Good dynamic properties allowed utilization of a high percentage of the equilibrium capacity at 10% breakthrough. For all proteins, a decreasing ratio of breakthrough to equilibrium capacity was detected with increasing feed concentration. This observation suggested a significant contribution of solid diffusion to the transport of proteins into the adsorbent particles. The specific architecture of the stationary phase, where the agarose base structure is derivatized with ion-exchange ligand-bearing dextran, may lead to this behavior.
Thömmes
J
J
Institut für Enzymtechnologie der Heinrich-Heine Universität Düsseldorf im Forschungszentrum Jülich, Stetternicher Forst, 52426 Jülich, Germany.
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange; D003911:Dextrans; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007782:Lactoglobulins / Q000302:isolation & purification; D009113:Muramidase / Q000302:isolation & purification; D010047:Ovalbumin / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification*; D012685:Sepharose; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D003911:Dextrans; D007074:Immunoglobulin G; D007782:Lactoglobulins; D011506:Proteins; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D012685:Sepharose; D009113:Muramidase