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Designed amino acid feed in improvement of production and quality targets of a therapeutic monoclonal antibody

Torkashvand, F ; Sharif University of Technology | 2015

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  1. Type of Document: Article
  2. DOI: 10.1371/journal.pone.0140597
  3. Publisher: Public Library of Science , 2015
  4. Abstract:
  5. 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. © 2015 Torkashvand et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
  6. Keywords:
  7. Amino acid ; Arginine ; Aspartic acid ; bevacizumab ; Glutamic acid ; Glycine ; Monoclonal antibody ; Amino acid analysis ; Antibody titer ; Cell growth ; CHO cell line ; Concentration (parameters) ; Controlled study ; Culture optimization ; Drug synthesis ; Factorial design ; Molecular weight ; Process optimization ; Productivity ; Quality control ; Response surface method
  8. Source: PLoS ONE ; Volume 10, Issue 10 , October , 2015 ; 19326203 (ISSN)
  9. URL: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0140597