Membrane affinity chromatography is an innovative technique that is receiving increasing attention by the biotechnology industry. Membrane chromatography is not limited by diffusion as the process based on resin beads, since the main transport mechanism is due to convection through the porous structure; therefore it is particularly suitable for purifying large bio-molecules such as IgG. In the present work the purification of immunoglobulin G using novel affinity membranes endowed with improved performance is discussed. The membrane adsorbers studied derive from the functionalization of epoxy membranes with natural and synthetic affinity ligands that show high specificity towards IgG. The resulting affinity membranes are fully characterized in complete adsorption, washing and elution affinity cycles. The separation performance of each affinity support has been determined by feeding both pure IgG solutions and a cell culture supernatant. The relevant process parameters, like maximum adsorption capacity, affinity equilibrium constant and selectivity, are compared for the different affinity membranes tested, as well as for available commercial membrane adsorbers. Exam of the impact of the improved affinity materials on industrial scale applications is also addressed. The experimental data collected have been used also for the validation of a simulation model proposed. The model developed is based on species mass balance equation over the membrane column, coupled with a suitable kinetic equation which represents the interaction between the IgG target molecule and the ligand immobilized on the porous support. Model simulations are in good agreement with the experimental affinity cycles, demonstrating the accuracy of the model to describe the transport phenomena in the column and the adsorption binding mechanism. On the basis of parameter values obtained for pure IgG solutions the model is able to predict the behaviour observed with a cell culture supernatant.
S. Dimartino, C. Boi, G.C. Sarti (2010). Experimental evaluation and theoretical analysis of affinity membrane adsorbers. s.l : s.n.
Experimental evaluation and theoretical analysis of affinity membrane adsorbers
DIMARTINO, SIMONE;BOI, CRISTIANA;SARTI, GIULIO CESARE
2010
Abstract
Membrane affinity chromatography is an innovative technique that is receiving increasing attention by the biotechnology industry. Membrane chromatography is not limited by diffusion as the process based on resin beads, since the main transport mechanism is due to convection through the porous structure; therefore it is particularly suitable for purifying large bio-molecules such as IgG. In the present work the purification of immunoglobulin G using novel affinity membranes endowed with improved performance is discussed. The membrane adsorbers studied derive from the functionalization of epoxy membranes with natural and synthetic affinity ligands that show high specificity towards IgG. The resulting affinity membranes are fully characterized in complete adsorption, washing and elution affinity cycles. The separation performance of each affinity support has been determined by feeding both pure IgG solutions and a cell culture supernatant. The relevant process parameters, like maximum adsorption capacity, affinity equilibrium constant and selectivity, are compared for the different affinity membranes tested, as well as for available commercial membrane adsorbers. Exam of the impact of the improved affinity materials on industrial scale applications is also addressed. The experimental data collected have been used also for the validation of a simulation model proposed. The model developed is based on species mass balance equation over the membrane column, coupled with a suitable kinetic equation which represents the interaction between the IgG target molecule and the ligand immobilized on the porous support. Model simulations are in good agreement with the experimental affinity cycles, demonstrating the accuracy of the model to describe the transport phenomena in the column and the adsorption binding mechanism. On the basis of parameter values obtained for pure IgG solutions the model is able to predict the behaviour observed with a cell culture supernatant.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
