Evaluation of membranes for use in on-line cell separation during mammalian cell perfusion processes.
In this study two microporous hollow fibre membranes were evaluated for their use as cell retention device in continuous perfusion systems. A chemically modified permanent hydrophillic PTFE membrane and a hydrophilized PP membrane were tested. To investigate the filtration characteristic under process conditions each membrane was tested during a long term perfusion cultivation of a hybridoma cell line. In both cultivations the conditions influencing membrane filtration (e.g. transmembrane flux) were kept constant. Filtration behaviour was investigated by monitoring transmembrane pressure and protein permeability. Transmembrane pressure was measured on-line with an autoclavable piezo-resistive pressure sensor. Protein permeability was determined by quantitative evaluation of unreduced, Coomassie stained SDS-PAGE. The membrane fouling process influences the filtration characteristic of both membranes in a different way. After fermentation the PP membrane was blocked by a thick gel layer located in the big outer pores of the asymmetric membrane structure. The hydraulic resistance was higher but the protein permeability was slightly better than of the PTFE membrane. For this reason the PP membrane should be preferred. On the other hand, transmembrane pressure decreases slower when the PTFE membrane is used, which favours this membrane for long term cultivations, especially when low molecular weight proteins (< 30 KD) are produced.
Büntemeyer
H
H
Institute for Cell Culture Technology, University of Bielefeld, Germany.
Böhme
C
C
Lehmann
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000302:isolation & purification*; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D006825:Hybridomas / Q000166:cytology*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000302:isolation & purification; D008322:Mammals; D008567:Membranes, Artificial; D051379:Mice; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D011138:Polytetrafluoroethylene; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D014168:Transferrin / Q000096:biosynthesis / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D007147:Immunoglobulin Light Chains; D008567:Membranes, Artificial; D011994:Recombinant Proteins; D014168:Transferrin; D011138:Polytetrafluoroethylene