Continuous production of large amounts of monoclonal immunoglobulins in hollow fibers using protein-free medium |
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Authors: | F. Dhainaut N. Bihoreau J. L. Meterreau J. Lirochon R. Vincentelli G. Mignot |
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Affiliation: | (1) Direction de la Recherche et du Développement, Innovation Therapeutique, CNTS, 3 Av. des Tropiques, BP 100, F-91943 Les Ulis, France |
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Abstract: | The performance of a protein-free medium was compared in culture flasks with a serum-supplemented medium and with a serum free medium in terms of cell growth and monoclonal antibody production by a murine hybridoma. We present results of continuous production in hollow fiber culture systems using serum-free medium and protein-free medium. In protein-free medium, it has been possible to produce large quantities of monoclonal antibody with a productivity similar to that obtained in serum-free medium. After a two steps purification process, monoclonal antibodies were characterized by SDS-PAGE, High Performance Size Exclusion Chromatography and Free Solution Capillary Electrophoresis. SDS-PAGE and high performance chromatography analysis have showed that purified monoclonal antibodies produced in serum-free medium or protein-free medium were similar. Furthermore, Capillary Electrophoresis characterization revealed that both MAbs were constituted by three isoforms with equivalent electrophoretic mobilities.Abbreviations CHES 2-(N-Cyclohexylamino)ethane-sulfonic acid - ECS Extracapillary Space - FSCE Free Solution Capillary Electrophoresis - HPSEC High Performance Size Exclusion Chromatography - ICS Intracapillary Space - MAb Monoclonal Antibody - PFM Protein-Free Medium - SFM Serum-Free Medium - SSM Serum-Supplemented Medium |
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Keywords: | continuous culture free solution capillary electrophoresis hollow fibers monoclonal antibody characterization protein-free medium |
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