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首页> 外文期刊>Biotechnology Progress >Screening of transformed insect cell lines for recombinant protein production.
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Screening of transformed insect cell lines for recombinant protein production.

机译：筛选用于重组蛋白生产的转化昆虫细胞系。

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Nine insect cell lines were evaluated for their potential as host systems for recombinant protein production using a new expression vector permitting the continuous high-level expression of secreted glycoproteins by transformed insect cells (Farrellet al., 1998). As a means of preliminary screening, all nine insect cell lines were transfected with the green fluorescence protein. Growth in static and suspension culture was then examined as a further method of screening. On the basis of their transfection efficiencies and cell growth characteristics, five insect cell lines, Bm5, High Five, IPLB-LdFB, IZD-MB-0503, and Sf-21, were selected for stable transformation to produce granulocyte-macrophage colony-stimulating factor (GM-CSF). These five celllines were stably transformed using an antibiotic resistance scheme and evaluated as a polyclonal population. Increasing the antibiotic concentration was found to cause not only a decrease in the specific growth rate but also an increase in the specificprotein production rate and final GM-CSF concentration. The transformed High Five cells exhibited by far the greatest specific protein production rate of 5.1X10-6 鎔/(cell.h), resulting in the highest final GM-CSF concentration of 22.8 mg/L when grown instatic culture. One cloned High Five cell line produced a GM-CSF concentration of 46 mg/L in static culture and 27 mg/L in suspension culture.
机译：使用一种新的表达载体，通过转化昆虫细胞连续高水平表达分泌的糖蛋白，评估了九种昆虫细胞系作为重组蛋白生产宿主系统的潜力（Farrellet等，1998）。作为初步筛选的一种方法，所有九种昆虫细胞系均用绿色荧光蛋白转染。然后检查静态和悬浮培养中的生长，作为另一种筛选方法。根据它们的转染效率和细胞生长特性，选择了五种昆虫细胞系Bm5，High Five，IPLB-LdFB，IZD-MB-0503和Sf-21进行稳定转化，以产生粒细胞-巨噬细胞集落因子（GM-CSF）。使用抗生素抗性方案将这五个细胞系稳定转化，并评估为多克隆群体。发现增加抗生素浓度不仅导致比生长速率的降低，而且导致比蛋白生产速率和最终GM-CSF浓度的提高。迄今为止，转化的高五细胞表现出最大的比蛋白产生速率5.1X10-6镕/(cell.h），在静态培养中生长时，最终的GM-CSF最高浓度为22.8 mg / L。一种克隆的High Five细胞系在静态培养中的GM-CSF浓度为46 mg / L，在悬浮培养中的GM-CSF浓度为27 mg / L。

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