title: Protein-free fed-batch culture of non-GS NS0 cell lines for production of recombinant antibodies.
abstract: Presented is a novel antibody production platform based on the fed-batch culture of recombinant, NS0-derived cell lines. A standardized fed-batch cell culture process was developed for five non-GS NS0 cell lines using enriched and optimized protein-free, cholesterol-free, and chemically defined basal and feed media. The process performed reproducibly and scaled faithfully from the 2-L to the 100-L bioreactor scale achieving a volumetric productivity of > 120 mg/L per day. Fed-batch cultures for all five cell lines exhibited significant lactate consumption when the cells entered the stationary or death phase. Peak and final lactate concentrations were low relative to a previously developed fed-batch process (FBP). Such low lactate production and high lactate consumption rates were unanticipated considering the fed-batch culture basal medium has an unconventionally high initial glucose concentration of 15 g/L, and an overall glucose consumption in excess of 17 g/L. The potential of this process platform was further demonstrated through additional media optimization, which has resulted in a final antibody concentration of 2.64 +/- 0.19 g/L and volumetric productivity of > 200 mg/L per day in a 13-day FBP for one of the five production cell lines. Use of this standardized protein-free, cholesterol-free NS0 FBP platform enables consistency in development time and cost effectiveness for manufacturing of therapeutic antibodies.
authors.affiliation: Process Sciences and Engineering, PDL BioPharma, Inc., 34700 Campus Drive, Fremont, California 94555, USA. john.burky@pdl.com
authors.forename: John E
authors.initials: JE
authors.lastname: Burky
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wesson
authors.forename: Amy
authors.initials: A
authors.lastname: Young
authors.forename: Sharyn
authors.initials: S
authors.lastname: Farnsworth
authors.forename: Ben
authors.initials: B
authors.lastname: Dionne
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhu
authors.forename: Taymar E
authors.initials: TE
authors.lastname: Hartman
authors.forename: Limin
authors.initials: L
authors.lastname: Qu
authors.forename: Weichang
authors.initials: W
authors.lastname: Zhou
authors.forename: Paul W
authors.initials: PW
authors.lastname: Sauer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D002784:Cholesterol / Q000494:pharmacology; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D002784:Cholesterol