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KMID : 1094720000050020130
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Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 2 p.130 ~ p.135
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Continuous stable production of von Willebrand Factor monoclonal antibody in spin filter bioreactor with bleeding technology
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Yun Jeong-Won
Lee Soo-Young
Choi Byung-Wook
Oh Han-Kyu
Kim Se-Ho
Byun Tae-Ho
Park Song-Yong
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Abstract
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The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1×107 cells/mL. When the perfusion rate was increased, the intermittent bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture exhibited higher physiological activity than that of the intermittent bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific vWF McAb productivity and cumulative production than those of the intermittent bleeding culture. Finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15 L bioreactor after one-month operation.
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KEYWORD
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cell culture, hybridoma, bleeding mode, perfusion, spin filter, monoclonal antibody
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