Spearman Maureen, Lodewyks Carly, Richmond Meika, Butler Michael
Dept. of Microbiology, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada.
Biotechnol Prog. 2014 May-Jun;30(3):584-93. doi: 10.1002/btpr.1930. Epub 2014 May 28.
Peptide hydrolysate supplements in mammalian cell cultures provide enhanced growth and productivity. The objective of this study was to compare the bioactivity of ten different commercially available hydrolysates from plant, microbial, and animal sources. The peptide hydrolysates were tested as supplements to cultures of Chinese hamster ovary (CHO) cells that produce human beta interferon (β-IFN). A soy hydrolysate was shown to support high cell growth but not protein productivity compared to an animal component hydrolysate (Primatone RL). On the other hand, a yeast hydrolysate showed lower cell growth, but comparable productivity of the recombinant protein. Glycosylation analysis showed that the glycan profile of β-IFN produced in yeast hydrolysate supplemented cultures was equivalent to that from Primatone RL-supplemented cultures. Fractionation of the yeast hydrolysate and Primatone RL produced a similar protein-assayed pattern except for one extra peak at around 1 kDa in the Primatone RL profile. A fraction taken at a molecular weight range of 1.5-1.7 kDa showed the highest growth promoting activity in both samples. However, four other fractions in yeast hydrolysate and two in Primatone RL at lower molecular weights showed some growth promoting activity. In conclusion, the yeast hydrolysates provided a good alternative to the animal sourced Primatone RL for high productivity of β-IFN from CHO cells with equivalent glycosylation.
哺乳动物细胞培养中的肽水解产物补充剂可提高细胞生长和生产力。本研究的目的是比较十种不同的市售植物、微生物和动物来源水解产物的生物活性。这些肽水解产物作为补充剂用于培养生产人β干扰素（β-IFN）的中国仓鼠卵巢（CHO）细胞。与动物成分水解产物（Primatone RL）相比，一种大豆水解产物可支持细胞的高生长，但不能提高蛋白质生产力。另一方面，一种酵母水解产物显示细胞生长较低，但重组蛋白的生产力相当。糖基化分析表明，在添加酵母水解产物的培养物中产生的β-IFN的聚糖谱与添加Primatone RL的培养物中的聚糖谱相当。酵母水解产物和Primatone RL的分级分离产生了相似的蛋白质检测模式，只是Primatone RL图谱中在约1 kDa处有一个额外的峰。在分子量范围为1.5-1.7 kDa处获取的一个级分在两个样品中均显示出最高的促生长活性。然而，酵母水解产物中其他四个较低分子量的级分和Primatone RL中的两个级分也显示出一定的促生长活性。总之，酵母水解产物为动物来源的Primatone RL提供了一个很好的替代品，可用于从CHO细胞中高效生产具有同等糖基化的β-IFN。
