title: Insight into the roles of hypoxanthine and thymidine [corrected] on cultivating antibody-producing CHO cells: cell growth, antibody production and long-term stability.
abstract: The potential of hypoxanthine and thymidine (H&T) to promote growth of CHO cells and production of monoclonal antibody (mAb) was explored in this study. It was demonstrated that H&T stimulated the initial cell growth and enhanced volumetric production of anti-human CD20 mAb by 22%, mainly through the elevated integrated viable cell concentration (IVCC). The moderate alteration in cell cycle distribution might partially account for the increased cell growth. Subsequent long-term stability studies indicated that H&T did not accelerate decay kinetics in mAb productivity. Specifically, cells under both nucleic acids-replete (H&T supplementation) and nucleic acids-hungry (methotrexate treatment) culture conditions showed similar stable mAb production during the first 2 months, followed by a gradual decline in the specific production rate (q(mAb)) with a 40% drop at the fourth month. In addition, the decreased transcript level of intracellular heavy chain (HC) of anti-human CD20 mAb correlated well with the decreased q(mAb). Furthermore, genomic mutation rate regarding the loss-of-function occurrence of the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene was determined, showing that H&T repressed the HPRT spontaneous mutation rate while methotrexate (MTX) provoked the mutation rate. Collectively, our data illustrated that H&T as potential medium additives promoted both initial cell growth and volumetric production of mAb, while not affecting the long-term stability of antibody-producing CHO cells.
authors.affiliation: The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.
authors.forename: Fei
authors.initials: F
authors.lastname: Chen
authors.forename: Li
authors.initials: L
authors.lastname: Fan
authors.forename: Jiaqi
authors.initials: J
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhou
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Ye
authors.forename: Liang
authors.initials: L
authors.lastname: Zhao
authors.forename: Wen-Song
authors.initials: WS
authors.lastname: Tan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D042822:Genomic Instability; D006801:Humans; D019271:Hypoxanthine / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D003470:Culture Media; D011994:Recombinant Proteins; D019271:Hypoxanthine; D013936:Thymidine