Departamento de Engenharia Química, Universidade Federal de São Carlos, Via Washington Luis, Km 235, CEP 13565-905, Sao Carlos, SP, Brazil.
Cytotechnology. 2008 May;57(1):67-72. doi: 10.1007/s10616-008-9134-3. Epub 2008 Feb 24.
Culture conditions that affect product quality are important to the successful operation and optimization of recombinant protein production. The objective of this study was to optimize culture conditions for growth of recombinant Drosophila melanogaster S2 cells (S2AcRVGP) in order to enhance the production of rRVGP. The addition of DMSO and glycerol to the medium and growth at a reduced temperature (22 degrees C) were the culture condition variations selected to be tested. Experimental cultures were first performed in serum-free Sf900 II medium in 250 ml Schott flasks. The most promising conditions identified in these experiments were also tested on a higher scale in a 3l bioreactor. In the Schott flasks experiments, all the changes in culture conditions resulted in an increase of rRVGP production. The protein concentration was 3.6-fold higher with addition of 1% DMSO and 1% glycerol and 9.3-fold higher when the cells were cultured at 22 degrees C instead of the standard 28 degrees C. The maximum concentration of rRVGP reached was 591 mug l(-1). In bioreactor experiments, with control of pH at 6.20 and DO at 50%, the reduced culture temperature (22 degrees C) was the strategy that promoted the highest glycoprotein production, 928 mug l(-1).
影响产物质量的培养条件对于重组蛋白生产的成功操作和优化非常重要。本研究的目的是优化重组黑腹果蝇 S2 细胞（S2AcRVGP）生长的培养条件，以提高 rRVGP 的产量。选择添加 DMSO 和甘油的培养基和在较低温度（22°C）下生长作为要测试的培养条件变化。实验培养首先在无血清 Sf900 II 培养基中在 250ml Schott 摇瓶中进行。在这些实验中确定的最有前途的条件也在 3L 生物反应器中进行了更高规模的测试。在 Schott 摇瓶实验中，所有培养条件的变化均导致 rRVGP 产量增加。添加 1% DMSO 和 1%甘油时，蛋白浓度提高了 3.6 倍，而在 22°C 而非标准 28°C 下培养时，蛋白浓度提高了 9.3 倍。达到的最大 rRVGP 浓度为 591μg l(-1)。在生物反应器实验中，在 pH 控制在 6.20 和 DO 控制在 50%的情况下，降低培养温度（22°C）是促进糖蛋白产量最高的策略，达到 928μg l(-1)。
