| Project/Area Number |
05555220
|
|---|
| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
生物・生体工学
|
|---|
| Research Institution | Nagoya University |
|---|
Principal Investigator |
KOBAYASHI Takeshi Nagoya University Faculty of Engineer., Dept.of Biotechnol., Prof., 工学部, 教授 (10043324)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
MIZUTANI Satoru Kirin Brewing Co.Ltd., Researcher, 研究員
UOZUMI Nobuyuki Nagoya University Faculty of Engineer., Dept.of Biotechnol., Assist.Prof., 工学部, 助手 (40223515)
本多 裕之 名古屋大学, 工学部, 助教授 (70209328)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Project Status |
Completed (Fiscal Year 1994)
|
| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1994: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1993: ¥2,900,000 (Direct Cost: ¥2,900,000)
|
|---|
| Keywords | Mammalian cell culture / Immobilization / Dissolved oxygen concentration / Nutrients concentrations / Control of culture / Erythropoietin / L-929 cell / vero細胞 / SV40 / 温度感受性 / 代謝制御 |
|---|
| Research Abstract |
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.
|
