Study on Increasing the Protein Expression of Cells by Specific Nutrient Compensation and Metabolism Control Strategy / 中国生物工程杂志

ABSTRACT

To increase the productivity and yield of recombinant protein in continual perfusion processing, Every amino acid consumption rate in continual perfusion culture of engineering CHO cell line which expressed recombinant TNFRp75: Fc fusion protein were analyzed. Then rational amino acids were accordingly added to improve its comprehensive utilizing. At the same time, glucose supply was controlled to make the concentration of glucose below 0.5 g/L for ameliorating the toxicity of lactate accumulation in order to decrease the perfusion rate. The result showed that the productivity of recombinant protein was 3.1 times (388mg/L) and the total yield was 4. 7 times (244. 4g) that of control cultures after nutrient compensation and metabolism control in 30 liter working volume, and the fermentation period was prolonged one week longer. The sialic acid content and bioactivity in vitro of recombinant TNFRp75: Fc were not changed after nutrient compensation and glucose control supply. Nutrient compensating and metabolic control in continual perfusion fermentation could significantly increase the productivity and yield of recombinant TNFRp75:Fc, and thus reduced relative industrialization costs.