title: Designed Amino Acid Feed in Improvement of Production and Quality Targets of a Therapeutic Monoclonal Antibody.
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.
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Torkashvand
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Behrouz
authors.initials: B
authors.lastname: Vaziri
authors.affiliation: Process Development Department, Aryogen Biopharma Inc., Alborz, Iran.
authors.forename: Shayan
authors.initials: S
authors.lastname: Maleknia
authors.affiliation: Department of Chemical & Petroleum Engineering, Biochemical & Bioenvironmental Research Center Sharif University of Technology, Tehran, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Heydari
authors.affiliation: Department of Chemical & Petroleum Engineering, Biochemical & Bioenvironmental Research Center Sharif University of Technology, Tehran, Iran.
authors.forename: Manouchehr
authors.initials: M
authors.lastname: Vossoughi
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Davami
authors.affiliation: Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Fereidoun
authors.initials: F
authors.lastname: Mahboudi
mesh_terms: D000596:Amino Acids / Q000494:pharmacology*; D000704:Analysis of Variance; D000818:Animals; D000068258:Bevacizumab / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D005818:Genetic Engineering; D007074:Immunoglobulin G / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism
chemical_list: D000596:Amino Acids; D003470:Culture Media; D007074:Immunoglobulin G; D011134:Polysaccharides; D000068258:Bevacizumab