效应面法优化CHO细胞培养基中几种主要成分的含量

Response Surface Methodology to Optimize the Content of Several Major Components of the CHO Cell Culture Medium

目的:考察培养基中葡萄糖、谷氨酰胺、血清、碳酸氢钠含量对CHO细胞生长繁殖的影响。方法:在CHO细胞培养基中添加不同成分的葡萄糖、谷氨酰胺、血清、碳酸氢钠,通过单因素实验结果结合Box-Behnken效应面法,根据二次回归模型的分析结果,以细胞表达蛋白体外活性为指标进行实验,考察培养基中葡萄糖、谷氨酰胺、血清、碳酸氢钠含量对细胞生长繁殖的影响。结果:根据回归方程分析结果,作出相应的曲面图和等高线图,优选出培养基中各组分的最佳配比为:葡萄糖2.54 g/L、谷氨酰胺0.59 g/L、血清8.3%,碳酸氢钠2.96 g/L。结论:Box-Behnken实验设计法用于细胞培养过程中考察培养基中各组分的优选是可行的,数学模型的预测值与实验观察值相符。通过对CHO细胞培养基成分的优化,使CHO细胞蛋白表达量高,有利于提高产品质量和降低生产成本。

Objective： To investigate the effect of glucose, glutamine, serum and sodium bicarbonate in the medium on CHO cell growth and multiplication. Methods： Different components of glucose, glutamine, serum and sodium bicarbonate were added into CHO cell culture medium, by combining the single factor experimental result and Box-Behnken response surface method, based on the analytical result of quadratic regression model, and taking cells expression of protein in vitro activity as Indices, experiments were carried out to investigate the effects of the glucose, glutamine, serum and sodium bicarbonate in the medium to the growth and reproduction of cell. Results： According to the analytical result of regression equation, made the corresponding kyrtograph and contour map, and selected out an optimum proportion of medium components： glucose 2.54 g/L, glutamine, 0.59 g/L, serum 8.3 % sodium bicarbonate 2.96 g/L. Conclusion： It is feasible to study preferably each component in the cell culture medium by using Box-Behnken experimental design method, and the values predicted by mathematical model is agree with the experimental observatio, the optimizing components of CHO cell culture medium not only improve the levels of protein expression in CHO cells, but also improve the quality of products and reduce the costs of products.