title: Effects of solution environment on mammalian cell fermentation broth properties: enhanced impurity removal and clarification performance.
abstract: The processing of recombinant proteins from high cell density, high product titer cell cultures containing mammalian cells is commonly performed using tangential flow microfiltration (MF). However, the increased cellular debris present in these complex feed streams can prematurely foul the membrane, adversely impacting MF capacity and throughput. In addition, high cell density cell culture streams introduce elevated levels of process-related impurities, which increase the burden on subsequent purification operations to remove these complex media components and impurities. To address this challenge, an evaluation of mammalian cell culture broth buffer properties was examined to determine if enhanced impurity removal and clarification performance could be achieved. A framework is presented here for establishing optimized mammalian cell culture buffer conditions, involving trade-offs between product recovery and purification and improved clarification at manufacturing-scale production. A reduction in cell culture broth pH to 4.7-5.0 induced flocculation and impurity precipitation which increased the average feed particle-size. These conditions led to enhanced impurity removal and improved MF throughput and filter capacity for several mammalian systems. Feed conditions were further optimized by controlling ionic composition along with pH to improve product recovery from high cell density/high product titer cell cultures.
authors.affiliation: Process Biochemistry Technical Development Biogen Idec 5200 Research Place, San Diego, California, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Westoby
authors.forename: James
authors.initials: J
authors.lastname: Chrostowski
authors.forename: Philippe
authors.initials: P
authors.lastname: de Vilmorin
authors.forename: John Paul
authors.initials: JP
authors.lastname: Smelko
authors.forename: Jonathan K
authors.initials: JK
authors.lastname: Romero
mesh_terms: D000818:Animals; D019149:Bioreactors; D002021:Buffers; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification*
chemical_list: D002021:Buffers; D003470:Culture Media; D007074:Immunoglobulin G; D011994:Recombinant Proteins