Proteome analysis of animal cells for development of efficient serum-free culture technology
动物细胞蛋白质组分析,用于开发高效的无血清培养技术
基本信息
- 批准号:14550771
- 负责人:
- 金额:$ 2.3万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2002
- 资助国家:日本
- 起止时间:2002 至 2003
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
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.
动物细胞的无血清培养对于临床和诊断上重要的蛋白质的大规模生产是必不可少的。为了开发高效的无血清培养技术,我们对中国仓鼠卵巢(CHO)细胞进行了蛋白质组学分析,CHO细胞被广泛用作生产复杂重组蛋白的宿主细胞。通过将维持在补充有10%胎牛血清(FBS)的培养基中的细胞逐步适应于含有较低浓度血清的培养基并最终适应于无血清培养基,建立能够在无血清培养基中生长的CHO细胞。测定了适应于无血清培养生长的细胞的比生长速率和比重组蛋白生产速率,发现其与维持在FBS补充培养基中的CHO细胞的比生长速率和比重组蛋白生产速率相当。然后通过细胞内蛋白质的二维凝胶电泳来研究血清非依赖性细胞,其中在第一维中具有固定的pH梯度(pH 3-10),在第二维中具有线性丙烯酰胺梯度(8-18%T)。将所得蛋白质表达模式与血清依赖性细胞中的蛋白质表达模式进行比较。在分析的700多个蛋白质点中,有几个蛋白质点显示出明显的增加或减少,表明这些蛋白质可能在无血清培养中的生长中起重要作用。
项目成果
期刊论文数量(9)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Hideki, Yamaji, Daigo, Hirakawa, Shin-ichi, Tagai, Hideki, Eukuda: "Production of protein kinase C-δ by the baculovirus-insect cell system in serum-supplemented and serum-free media"Journal of Bioscience and Bioengineering. 95-2. 185-187 (2003)
Hideki、Yamaji、Daigo、Hirakawa、Shin-ichi、Tagai、Hideki、Eukuda:“杆状病毒-昆虫细胞系统在添加血清和无血清的培养基中生产蛋白激酶 C-δ”《生物科学与生物工程杂志》95。 -2。185-187(2003)
- DOI:
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- 影响因子:0
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- 通讯作者:
Chisa, Hayashi, Kentaro, Sakai, Hideki, Yamaji, Hideki, Fukuda: "Comparative study on delivery of phosphatidic acid to serum-free culture of Chinese hamster ovary cells"Journal of Bioscience and Bioengineering. 96-2. 196-198 (2003)
Chisa、Hayashi、Kentaro、Sakai、Hideki、Yamaji、Hideki、Fukuda:“向中国仓鼠卵巢细胞无血清培养物输送磷脂酸的比较研究”生物科学与生物工程杂志。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Hideki Yamaji: "Production of protein kinase C-δ by the baculovirus-insect cell system in serum-supplemented and serum-free media"Journal of Bioscience and Bioengineering. 95. 185-187 (2003)
Hideki Yamaji:“杆状病毒昆虫细胞系统在补充血清和无血清培养基中生产蛋白激酶 C-δ”《生物科学与生物工程杂志》95. 185-187 (2003)。
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- 发表时间:
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- 影响因子:0
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- 通讯作者:
Norikatsu Nishikawa: "Recombinant protein production by the baculovirus-insect cell system in basal media without serum supplementation"Cytotechnology. (未定). (2004)
Norikatsu Nishikawa:“在不添加血清的基础培养基中通过杆状病毒-昆虫细胞系统生产重组蛋白”(待定)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Chisa Hayashi: "Comparative study on delivery of phosphatidic acid to serum-free culture of Chinese hamster ovary cells"Journal of Bioscience and Bioengineering. 96. 196-198 (2003)
Chisa Hayashi:“向中国仓鼠卵巢细胞无血清培养物输送磷脂酸的比较研究”生物科学与生物工程杂志。
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YAMAJI Hideki其他文献
YAMAJI Hideki的其他文献
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{{ truncateString('YAMAJI Hideki', 18)}}的其他基金
Secretory production of a functional membrane protein
功能性膜蛋白的分泌产生
- 批准号:
24656505 - 财政年份:2012
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Efficient production of virus-like particles using insect cells
利用昆虫细胞高效生产病毒样颗粒
- 批准号:
22360345 - 财政年份:2010
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Continuous production of virus-like particles by recombinant insect cells
重组昆虫细胞连续生产病毒样颗粒
- 批准号:
19560777 - 财政年份:2007
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Display of functional antibody Fab fragment on the baculovirus surface
杆状病毒表面功能性抗体Fab片段的展示
- 批准号:
17560690 - 财政年份:2005
- 资助金额:
$ 2.3万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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