Recombinant antibody production by perfusion cultures of rCHO cells in a depth filter perfusion system.
Recombinant Chinese hamster ovary cells, producing recombinant antibody against the human platelet, were cultivated in a depth filter perfusion system (DFPS). When perfusion cultures with working volume of 1 L were operated at perfusion rates of 5/d and 6/d, volumetric antibody productivities reached values 28 and 34 times higher than that of batch suspension culture in Erlenmeyer flasks and 43 and 53 times higher than that of batch culture in a controlled stirred tank reactor, respectively. Perfusion cultures in the DFPS showed stable antibody production over the whole culture period of up to 20 days. In the DFPS, inoculated cells in suspension were entrapped in a few hours within the depth filter matrix by medium circulation and retained there until the void space of the filter matrix was saturated by the cultured cells. After cells in the depth filter matrix reached saturation, overgrown viable cells at a perfusion rate of 5/d or 6/d were continuously collected into waste medium at a density of 2-4 x 10(5) cells/mL, which resulted in stable operation at high perfusion rates, maintaining values of process parameters such as glucose/lactate concentration, pH, and dissolved oxygen concentration. Because the DFPS overcomes most drawbacks observed with conventional perfusion systems, it is preferable to be used as a key culture system to produce monoclonal antibody stably for a long culture period.
Lee
Joon Chul
JC
Department of Bioscience and Biotechnology, Sejong University, 98 Gunja-dong, Gwangjin-gu, Seoul 143-747, Korea.
Chang
Ho Nam
HN
Oh
Duk Jae
DJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D006224:Cricetinae; D003470:Culture Media; D005374:Filtration / Q000295:instrumentation; D010477:Perfusion / Q000295:instrumentation* / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors
D000911:Antibodies, Monoclonal; D003470:Culture Media; D011994:Recombinant Proteins