Abstract
Cell culture feeds optimization is a critical step in process development of pharmaceutical recombinant protein production. Amino acids are the basic supplements of mammalian cell culture feeds with known effect on their growth promotion and productivity. In this study, we reported the implementation of the Plackett-Burman (PB) multifactorial design to screen the effects of amino acids on the growth promotion and productivity of a Chinese hamster ovary DG-44 (CHO-DG44) cell line producing bevacizumab. After this screening, the amino acid combinations were optimized by the response surface methodology (RSM) to determine the most effective concentration in feeds. Through this strategy, the final monoclonal antibody (mAb) titre was enhanced by 70%, compared to the control group. For this particular cell line, aspartic acid, glutamic acid, arginine and glycine had the highest positive effects on the final mAb titre. Simultaneously, the impact of the designed amino acid feed on some critical quality attributes of bevacizumab was examined in the group with highest productivity. The product was analysed for N-glycan profiles, charge variant distribution, and low molecular weight forms. The results showed that the target product quality has been improved using this feeding strategy. It was shown how this strategy could significantly diminish the time and number of experiments in identifying the most effective amino acids and related concentrations in target product enhancement. This model could be successfully applied to other components of culture media and feeds.
Highlights
Recent progress in cell culture technology for recombinant CHO cells has led to substantial enhancements in target protein’s production [1, 2]
We reported the implementation of the Plackett-Burman (PB) multifactorial design to screen the effects of amino acids on the growth promotion and productivity of a Chinese hamster ovary
No generic procedure exists for cell culture medium and feed optimization, and reports on the optimization of culture media or feeding strategies for CHO cell cultures are limited in number
Summary
Recent progress in cell culture technology for recombinant CHO cells has led to substantial enhancements in target protein’s production [1, 2] This advancement in the production yields is mostly due to the extension of stable high producers through vector design and host cell engineering methods, as well as medium optimization and process development [3,4,5]. A valuable start to optimizing the medium or feed is to focus on basic groups of ingredients composing mammalian cell culture media Traditional optimization methods, such as the titration of single components, are reliable, but can be labour intensive and time consuming [7,8,9]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
