Membrane chromatography is a purification technique that is receiving increasing attention by the biotechnology industry. However, the industrial application of this process has been limited by the relative low binding capacity of membranes with respect to chromatography beads. In this work the purification of IgG using novel affinity membranes endowed with improved performances will be discussed. The membranes have been prepared using a new family of pre-activated epoxy membranes functionalized with natural and synthetic affinity ligands that show high specificity towards the Fc portion of antibodies. The resulting affinity membranes have been fully characterized in batch and dynamic experiments with both pure IgG solutions and a cell culture supernatant containing monoclonal IgG1. The influence of membrane structure parameters and ligand density on separation performance has been investigated to draw indications for material improvements. The relevant process parameters like the maximum binding capacity, the affinity equilibrium constant and selectivity have been obtained in complete adsorption, washing and elution chromatographic cycles; the results for the different affinity membranes tested were critically analyzed and compared with data of commercially available membrane adsorbers. Considerations about the application of the improved affinity materials in industrial scale will be addressed.

Novel affinity membranes for IgG purification

BOI, CRISTIANA;DIMARTINO, SIMONE;SARTI, GIULIO CESARE
2009

Abstract

Membrane chromatography is a purification technique that is receiving increasing attention by the biotechnology industry. However, the industrial application of this process has been limited by the relative low binding capacity of membranes with respect to chromatography beads. In this work the purification of IgG using novel affinity membranes endowed with improved performances will be discussed. The membranes have been prepared using a new family of pre-activated epoxy membranes functionalized with natural and synthetic affinity ligands that show high specificity towards the Fc portion of antibodies. The resulting affinity membranes have been fully characterized in batch and dynamic experiments with both pure IgG solutions and a cell culture supernatant containing monoclonal IgG1. The influence of membrane structure parameters and ligand density on separation performance has been investigated to draw indications for material improvements. The relevant process parameters like the maximum binding capacity, the affinity equilibrium constant and selectivity have been obtained in complete adsorption, washing and elution chromatographic cycles; the results for the different affinity membranes tested were critically analyzed and compared with data of commercially available membrane adsorbers. Considerations about the application of the improved affinity materials in industrial scale will be addressed.
2009
2009 AIChE Annual Meeting and Fall Showcase Conference Proceedings on CD
1
1
C. Boi; S. Dimartino; G. Sarti
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/86477
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