不同通气搅拌条件下体外培养CHO细胞高表达西妥昔单抗的研究

Study on High Level Expression of Cetuximab in Vitro Culture by Different Ventilationand Stirring Conditions in CHO Cells

摘要:

以稳定表达西妥昔单抗的CHO细胞为研究对象，采用无血清悬浮培养方式，建立4种不同的通气及搅拌方式，通过对细胞密度、细胞活率以及蛋白浓度进行跟踪检测，考察其对细胞生长和蛋白表达水平的影响。实验结果表明新型通气搅拌方式的细胞密度和蛋白表达量优于或相当于传统的微泡通气方式，同时还改善了细胞培养的微环境，且在清洗验证上具有很大优势，为大泡通气在动物细胞培养的应用方面奠定了一定的基础，也为优化中试规模制备西妥昔的CHO细胞培养工艺奠定了基础。

Abstract:

Four different Ventilation and stirring conditions were established , Taking CHO cells expressed Cetuximab as research object and adopting the serum-free mediumand suspension culture, the effect of different Ventilation conditions on cell density、cell viability and protein concentration were contrasted. The results show that compared with traditional Ventilation like micro bubble and no bubble, cell density and protein concentration of big bubble Ventilation was almost the same even better in some respects. The new aeration not only changed the microenvironment of the CHO cells, but also had a great advantage in the cleaning validation. This Study laid a foundation for the application of big bubble ventilation on Large-scale Mammalian Cell Culture, and also laid a foundation for the optimization of CHO culture technology in pilot scale.