Bioresour Bioprocess
作者: 发布时间:2016-03-25 14:03:00
Bioresources and Bioprocessing 2016, 3:5
http://bioresourcesbioprocessing.springeropen.com/articles/10.1186/s40643-015-0072-6
进料培养基中不同谷氨酰胺:天冬酰胺的比率对CHO-DHFR细胞反应的细胞生长、抗体产量、代谢废物、谷氨酸盐及能量代谢的影响
Li-xiang Zhang, Wei-yan Zhang, Chen Wang, Jin-tao Liu, Xian-cun Deng, Xu-ping Liu, Li Fan and Wen-song Tan
East China University of Science and Technology, Peoples’ Republic of China
【摘要】背景:优化化学成分明确的培养基一直以来是生产单克隆抗体的重要途径。一般加入谷氨酰胺:天冬酰胺的比例为2:1。研究表明,天冬酰胺对抗体生产阶段很重要。在补料培养基中提高天冬酰胺:谷氨酰胺的比率可能是在CHO-DHFR的细胞中提高抗体生产量的有效方法。结果:我们优化了补料培养基中两种重要氨基酸的总量和比例以提高抗体产量。提高天冬酰胺:谷氨酰胺(FB-H)的比率可以在到达静止期后提高细胞密度、改善细胞的维持、增加抗体产量。代谢通量分析表明,谷氨酰胺:天冬酰胺的比例对糖酵解的影响不大。高天冬酰胺:谷氨酰胺(FB-H)比的TCA循环提高20%,能量代谢提高22.6%,生成更少的副产物乳酸盐。结论:在不影响细胞生长的前提下,应适当提高补料培养基中的天冬酰胺:谷氨酰胺的比例,以实现单抗的高产目标。
Metabolic flux distribution. a The calculated values of flux were normalized with respect to that of the low proportion between asparagine and glutamine. The line size was increased with the value of flux. The figure indicated that TCA cycle was strengthened in high proportion condition. b Metabolism of high proportion between glutamine and asparagine in CHO-DHFR cell lines. Glutamate and aspartate tended to flow into TCA cycle for immediate metabolites and energy. c Metabolism of low proportion between glutamine and asparagine in CHO-DHFR cell lines. Glutamate was inclined to produce lactate via a truncate TCA cycle