| Project/Area Number |
14550771
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Kobe University |
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Principal Investigator |
YAMAJI Hideki Kobe Univ., Faculty of Engineering, Research Associate, 工学部, 助手 (40283874)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUDA Hideki Kobe Univ., Graduate School of Science and Technology, Professor, 自然科学研究科, 教授 (30263396)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | Cell culture / Animal cells / Insect cells / Serum-free culture / Recombinant protein production / Proteome analysis / CHO cells / CHO / Sf9 / 浮遊化 |
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| Research Abstract |
Serum-free culture of animal cells is essential for large-scale production of clinically and diagnostically important proteins. With the aim of developing efficient serum-free culture technology, we have undertaken a proteomic analysis of Chinese hamster ovary (CHO) cells, which are widely used as host cells in the production of complex recombinant proteins. CHO cells capable of growing in a serum-free medium were established by stepwise adaptation of cells maintained in a medium supplemented with 10% fetal bovine serum (FBS) to a medium containing lower concentrations of serum and finally to a serum-free medium. Specific growth rate and specific recombinant protein production rate of the cells adapted to growth in serum-free culture were measured and found to he comparable to those of CHO cells maintained in a FBS-supplemented medium. The serum-independent cells were then studied by two-dimensional gel electrophoresis of intracellular proteins with immobilized pH gradient (pH 3-10) in the first dimension and a linear acrylamide gradient (8-18%T) in the second dimension. The resulting protein expression patterns were compared with those in serum-dependent cells. Among over 700 protein spots analyzed, several spots showed marked increase or decrease, indicating that these proteins may play an important role in growth in serum-free culture.
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