表达乙型肝炎表面抗原的重组CHO细胞无血清培养基的优化及生物反应器培养  

Optimization of Serum-free Medium for Recombinant CHO Cells Expressing HBsAg and Culture in Bioreactor

作　　者：滕小锘[1] 易小萍[1] 孙祥明[1] 张元兴[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《中国生物制品学杂志》2010年第10期1080-1083,1086,共5页Chinese Journal of Biologicals

基　　金：国家科技重大专项(2009ZX09503-013);上海市重点学科建设项目资助(B505)

摘　　要：目的优化表达乙型肝炎表面抗原(HBsAg)的重组CHO细胞的无血清培养基,并进行生物反应器高密度培养。方法通过对现有重组CHO细胞无血清培养基SFMB的氨基酸、蛋白类激素、蛋白水解物等的优化,建立针对表达HBsAg的重组CHO细胞无血清低蛋白(<10mg/L)培养基SFMC。并利用此培养基在生物反应器中分批、流加、灌注悬浮培养重组CHO细胞,通过最大细胞密度和HBsAg产率评价不同的培养模式。结果 SFMC与原培养基SFMB相比,使重组CHO细胞的最大细胞密度提高了20%,HBsAg表达量提高了25%。在生物反应器培养过程中,灌注培养的重组CHO细胞表达的HBsAg产率为0.70mg/(L·d),较分批和流加培养的0.30mg/(L·d)提高了约133%。结论通过无血清培养基优化及生物反应器高密度培养,可显著提高重组CHO细胞表达HBsAg的效率。Objective To optimize a serum-free medium for high density culture of CHO cells expressing HBsAg in bioreac-tor. Methods SFMC, a low-protein（ 10 mg / L）medium for CHO cells expressing HBsAg, was developed by optimizing the compoone, then used for batch, fedbatch and perfused suspension cultures of recombinant CHO cells expressing HBsAg in bioreactor. The culture modes were evaluated by the maximum cell density and HBsAg yield. Results Compared with those culture in SFMB medium, the maximum density of recombinant CHO cells cultured in SFMC medium increased by 20%, while the HBsAg expression level increased by 25%. During culture in bioreactor, the HBsAg yield in CHO cells in perfused culture was 0. 70 mg /（L·d）, which increased by about 133% as compared with those in batch and fedbatch cultures[both 0. 30 mg /（L·d）]. Conclusion Optimization of serum-free medium and high density culture in bioreactor increased the expression efficacy of HBsAg in recombinant CHO cells.

关 键 词：CHO细胞 肝炎表面抗原 乙型 培养基 无血清 生物反应器 

分 类 号：Q813.11[生物学—生物工程] R392-33[医药卫生—免疫学]