Continuous culture of CHO-K1 cells producing thrombomodulin and estimation of culture conditions |
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| Institution: | 1. Production Technology Research Laboratories, Daiichi Pharmaceutical Co. Ltd., 16-13, Kitakasai 1-chome, Edogawa-ku, Tokyo 134, Japan;2. Molecular Biology Research Laboratory, Daiichi Pharmaceutical Co. Ltd., 16-13, Kitakasai 1-chome, Edogawa-ku, Tokyo 134, Japan;3. Toyobo Pharmaceuticals Research Center, Toyobo Co. Ltd., 2-1-1, Katata, Otsu, Shiga 520-02, Japan;4. Chemical Engineering Department, Kyoto University, Yoshida Hon-machi, Sakyo-ku, Kyoto 606, Japan;1. Centre for Bioprocess Engineering Research, Department of Chemical Engineering, University of Cape Town, Rondebosch 7701, South Africa;2. Royal School of Mines, Department of Earth Sciences and Engineering, Imperial College London, SW7 2AZ, UK;1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;2. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China;3. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China;4. Jiangsu Provincial Research Center for Bioactive Product Processing Technology, Jiangnan University, Wuxi 214122, China;5. China Agricultural Veterinary Biological Science and Technology Co., Ltd, Lanzhou 730046, China;1. Multiscale Bioengineering, Technical Faculty, Bielefeld University, Bielefeld, Germany;2. Center for Biotechnology (CeBiTec), Bielefeld University, Bielefeld, Germany;3. Microsystems in Bioprocess Engineering, Institute of Process Engineering in Life Sciences, Karlsruhe Institute of Technology, Karlsruhe, Germany |
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| Abstract: | Recombinant Chinese hamster ovary (CHO-K1) cells expressing human soluble thrombomodulin (rsTM) were cultured in a continuous culture system with a fluidized-bed reactor. Cells were grown in a medium containing 1% serum for 10 d, and then cultured in a serum-free medium. The protein production rate increased remarkably in the serum-free culture, with a decrease in the lactate production rate. This suggests that CHO-K1 cells exhibit different physiological characteristics in response to serum removal from the medium, which resulted in a higher rsTM concentration (about 60 mg/l). A procedure for estimating protein productivity was developed using experimental glucose and lactate measurements. In this procedure, cell density was estimated from the glucose consumption rate, and the specific protein (rsTM) production rate was obtained from the ratio of lactate production/glucose consumption (ΔL/ΔG). Since the cell density and protein productivity in repeated batch culture were well estimated, the procedure was applied to continuous culture in a fluidized-bed bioreactor culture. The estimation procedure was also found to be effective in this continuous culture using the models derived from the repeated batch culture. |
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