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温度和氧气对昆虫细胞培养中细胞生长及重组蛋白生产的影响

Effect of temperature and oxygen on cell growth and recombinant protein production in insect cell cultures.

作者信息

Reuveny S, Kim Y J, Kemp C W, Shiloach J

机构信息

Biotechnology Unit, LCDB, NIDDK, NIH, Bethesda, MD 20892.

出版信息

Appl Microbiol Biotechnol. 1993 Feb;38(5):619-23. doi: 10.1007/BF00182800.

Abstract

The effect of temperature and O2 saturation on the production of recombinant proteins beta-galactosidase and human glucocerebrosidase by Spodoptera frugiperda cells (Sf9) infected with recombinant Autographa californica nuclear polyhedrosis virus was investigated. The rates of cell growth, glucose consumption, O2 consumption and product expression were measured at temperatures between 22 degrees C and 35 degrees C. The results indicated that possible O2 limitation may be alleviated without compromising the maximum cell yield by lowering the incubation temperature from 27 degrees C to 25 degrees C. The expression level of the recombinant proteins at 27 degrees C was similar to that obtained at 22 degrees C and 25 degrees C; lower protein yields were obtained at 30 degrees C. An increase in temperature from 22 degrees C to 27 degrees C led to earlier production of the proteins and to an increase in the proportion of the product released outside the cells.

摘要

研究了温度和氧气饱和度对感染重组苜蓿银纹夜蛾核型多角体病毒的草地贪夜蛾细胞(Sf9)产生重组蛋白β-半乳糖苷酶和人葡糖脑苷脂酶的影响。在22℃至35℃之间的温度下测量细胞生长速率、葡萄糖消耗、氧气消耗和产物表达。结果表明,通过将培养温度从27℃降至25℃,可以缓解可能的氧气限制,而不会影响最大细胞产量。27℃时重组蛋白的表达水平与22℃和25℃时相似;30℃时蛋白质产量较低。温度从22℃升高到27℃导致蛋白质更早产生,并且细胞外释放的产物比例增加。

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