Re-use of spent cell culture medium in pilot scale and rapid preparative purification with membrane chromatography.
Based on experiments in bench scale, a recycling of spent cell culture medium was performed in a 100-1 pilot scale bioreactor. The cell cultivation has been done as a repeated batch procedure after the initial batch in the following four repeated batches spent medium from the previous batch was partially re-used. After microfiltration and ultrafiltration a part of the filtrate was mixed with a concentrate of amino acids and glucose, sterile filtered and subsequently filled back into the bioreactor. Up to 65% of the harvested cell- and product-free spent medium was re-used in each repeated batch. This procedure results in a saving of pure and waste water volume and saving of supplemented proteins as transferrin, insulin and lipoproteins and, therefore, also in a reduction of the production costs. A strongly acidic membrane ion exchanger was evaluated for the ability to purify the monoclonal antibodies from the pilot scale cultivation. Within minutes, gram quantities of product could be purified in a high flux system, especially developed for this purpose, achieving purities of 80%. The capacity of the acidic membrane ion exchanger was found in former investigations to be 1 mg cm-2 with recoveries up to 96%. Final purification was carried out by gel column filtration.
Riese
U
U
Institute for Cell Culture Technology, University of Bielefeld, Germany.
Lütkemeyer
D
D
Heidemann
R
R
Büntemeyer
H
H
Lehmann
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification; D001709:Biotechnology / Q000295:instrumentation; D002452:Cell Count; D002455:Cell Division; D002852:Chromatography, Ion Exchange / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media / Q000737:chemistry / Q000302:isolation & purification; D003584:Cytological Techniques / Q000295:instrumentation; D006825:Hybridomas / Q000166:cytology / Q000276:immunology; D008567:Membranes, Artificial; D051379:Mice; D010865:Pilot Projects
D000911:Antibodies, Monoclonal; D003470:Culture Media; D008567:Membranes, Artificial