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
滕小锘;易小萍;孙祥明;张元兴;
摘要(Abstract):
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。
关键词(KeyWords): CHO细胞;肝炎表面抗原,乙型;培养基,无血清;生物反应器
基金项目(Foundation): 国家科技重大专项(2009ZX09503-013);; 上海市重点学科建设项目资助(B505)
作者(Authors): 滕小锘;易小萍;孙祥明;张元兴;
DOI: 10.13200/j.cjb.2010.10.51.tengxn.008
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