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Growth of recombinant Drosophila melanogaster Schneider 2 cells producing rabies virus glycoprotein in bioreactor employing serum-free medium

机译:使用无血清培养基在生物反应器中生长产果蝇狂犬病毒糖蛋白的重组果蝇施耐德2细胞

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摘要

Drosophila melanogaster Schneider 2 (S2) cells have been increasingly used as a suitable expression system for the production of different recombinant proteins, and the employment of bioreactors for large-scale culture is an important tool for this purpose. In this work, Drosophila S2 cells producing the rabies virus glycoprotein RVGP were cultivated in bioreactor, employing a serum-free medium, aiming an improvement in cell growth and in glycoprotein production. To overcome cell growth limitation commonly observed in stirred flasks, different experiments in bioreactor were performed, in which some system modifications were carried out to attain the desired goal. The study showed that this cell line is considerably sensitive to hydrodynamic forces, and a high cell density (about 16.0 × 106 cells mL−1) was only obtained when Pluronic F68® percentage was increased to 0.6% (w/v). Despite ammonium concentration affected RVGP production, and also cell growth, an elevated amount of the target protein was obtained, attaining 563 ng 10−7 cells.
机译:果蝇Schneider 2(S2)细胞已越来越多地用作生产不同重组蛋白的合适表达系统,为此目的,采用生物反应器进行大规模培养是重要的工具。在这项工作中,使用无血清培养基在生物反应器中培养产生狂犬病病毒糖蛋白RVGP的果蝇S2细胞,旨在改善细胞生长和糖蛋白的产生。为了克服通常在搅拌烧瓶中观察到的细胞生长限制,在生物反应器中进行了不同的实验,其中进行了一些系统修改以达到所需的目标。研究表明,该细胞系对流体动力非常敏感,仅当Pluronic F68获得高细胞密度(约16.0×10 6 个细胞mL -1 )。 ®百分比增加到0.6%(w / v)。尽管铵浓度影响了RVGP的产生以及细胞的生长,但仍获得了高水平的靶蛋白,达到563ng 10 -7 细胞。

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