Production of useful biochemicals by immobilized animal cells

通过固定化动物细胞生产有用的生化物质

• 批准号: 63550723
• 负责人: IIJIMA Shinji
• 金额: $ 1.15万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63550723/
• 关键词: Animal cells; Hybridoma; Immobilization; Cellulose; Monoclonal antibody; Gel entrapment; Alginate; Bioreactor; 胞括固定; モノクローナル抗体

In order to improved the production of useful biochemicals by animal cell culture, a novel immobilization methods have been developed. Two hybridoma cell lines that produced anti- erythropoietin and anti-alpha -amylase monocional antibody were entrapped in alginate gel and then the immobilized animal cell gel was covered with urethane polymer. By this method of immobilization, more than 10^7 cells/ml gel was obtained and the volumetric productivity of the antibody increased almost 10 times comparing to the regular cultures.In order to culture anchorage depended cells, a novel macroporous cellulose fiber(SIC) was used. Transformed human endothelial cells were immobilized on this support and cultured in a fluidized bed reactor. By optimizing medium and oxygen supply, high concentration of cells more than 10 cells/ml. SIC could be obtained. By introducing a charged group on the surface of SIC, hybridoma cell was also immobilized and high concentration of cells near 10^8 cells/ml. SIC was attained.By regular cultivation of animal cells, oxygen supply is the most serious problem. Therefore, we compared several method and found that enough amount of oxygen for the growth of high concentration of cells more than 10^7 cell/ml could be supplied by using perfluorochemicals.

为了提高动物细胞培养有用生化物质的生产，开发了一种新的固定化方法。将产生抗促红细胞生成素和抗α-淀粉酶单克隆抗体的两种杂交瘤细胞系包埋在藻酸盐凝胶中，然后用氨基甲酸乙酯聚合物覆盖固定的动物细胞凝胶。通过这种固定化方法，获得了超过10^7个细胞/ml的凝胶，抗体的体积生产率比常规培养物增加了近10倍。为了培养贴壁依赖性细胞，一种新型的大孔纤维素纤维（SIC）被使用了。将转化的人内皮细胞固定在该支持物上并在流化床反应器中培养。通过优化培养基和氧气供应，细胞浓度高至10个/ml以上。可以获得SIC。通过在SIC表面引入带电基团，杂交瘤细胞也被固定化，细胞浓度接近10^8个/ml。达到SIC。 常规培养动物细胞，氧气供应是最严重的问题。因此，我们比较了几种方法，发现使用全氟化合物可以为超过10^7个细胞/ml的高浓度细胞的生长提供足够的氧气。

项目成果

Hitoshi,Yoshida: Journal of Chemical Engineering of Japan. (1989)

吉田仁：日本化学工程学报。

S.Iijima, T.Mano, M.Taniguchi and T.Kobayashi: "Immobilization of hybridoma cells with alginate and improved monoclonal antibody production" Appl. Microbiol. Biotechnol. 28(6), 572-576, 1988.

S.Iijima、T.Mano、M.Taniguchi 和 T.Kobayashi：“用藻酸盐固定杂交瘤细胞并改善单克隆抗体的产生”Appl。

Shinji,Iijima: Applied Microbiology and Biotechnology. 28(6). 572-576 (1988)

饭岛真司：应用微生物学和生物技术。

T.Mano, T.Kimura, S.Iijima, K.Takahashi, H.Takeuchi and T. Kobayashi: "Comparison oxygen supply methods for animal cell culture" J. Chem. Technol. Biotechnol.

T.Mano、T.Kimura、S.Iijima、K.Takahashi、H.Takeuchi 和 T. Kobayashi：“动物细胞培养供氧方法的比较”J. Chem。

Shinji Iijima: "Immobilization of hybridoma cells with alginate and improved monoclonal antibody production." Applied Microbiology and Biotechnology. 28(6). 572-576 (1988)

Shinji Iijima：“用藻酸盐固定杂交瘤细胞并改善单克隆抗体的生产。”