動物細胞の固定化及び培養環境の制御による有用物質の効率的生産

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1.A photo-crosslinkable resin, BIX12, was selected from among various photo-crosslinkable resins for the immobilization of animal cells. BIX12 had no cytotoxic effect on the growth of hybridoma cells and the production of monoclonal antibody. A high viable cell density of more than 10^7 cells/ml-gel was attained, and the antibody productivity was improved 8.5-fold compared with conventional suspension culture using a spinner flask.2.By introduction of the positive charge to macroporous cellulosic support, anchorage-independent hybridoma cells were immobilized more than 10-fold compared with the unmodified support. Large amount of monoclonal antibody was continuously produced for long term. Recombinant L-929 cells transfected with the human erythropoietin (EPO) gene were also immobilized in the macroporous cellulosic support. A high cell density of more than 2x10^7 cells/ml-support and high EPO productivity were achieved and maintained for 50 d through the use of the inner-loop type air-lift bioreactor.The effects of dissolved oxygen concentration (DO) on anchorage-dependent cell growth, cell metabolism, and erythropoietin (EPO) or tissue plasminogen activator (tPA) production were investigated. The optimal DO for cell growth was 5mg/l, whereas the optimal EPO or tPA production was observed at higher DO such as 15mg/l.4.An on-line system using HPLC was developed for the measurement of glucose, glutamine and lactate in a culture broth. Using the system, the glucose and glutamine concentrations were controlled simultaneously by an adaptive-control algorithm within the ranges 0.2-2.0 and 0.1-0.6g/L,respectively. When the glucose or glutamine concentration was controlled at the low level of 0.2g/L,antibody production increased two-fold compared to the high glucose or glutamine level.