title: pH-conductivity hybrid gradient cation-exchange chromatography for process-scale monoclonal antibody purification.
abstract: The commercial production of recombinant human monoclonal antibody therapeutics demands robust processes. In this article we describe the development of a pH-conductivity hybrid gradient for a cation-exchange chromatography step to obtain high binding capacity and consistent purification resolution in scale process. Operational parameters and their ranges were characterized with DOE statistical method. Aggregate, DNA and leached protein A removal were examined during development. The advantages and disadvantages of hybrid gradient elution compared to sodium chloride gradient elution were explored. As this step was designed as a good fit for the compatibility of the feed and operating pH/conductivity conditions for next step, the effects of elution by either changing sodium chloride concentration or changing pH of elution buffers on overall separation efficiency were compared. The operation condition was further confirmed in six 2000 L scale runs. The thorough evaluation demonstrated process reliability of hybrid gradient cation-exchange chromatography with high step purity and yield.
authors.affiliation: Department of Purification Process Development, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA. joez@amgen.com
authors.forename: Joe X
authors.initials: JX
authors.lastname: Zhou
authors.forename: Shinta
authors.initials: S
authors.lastname: Dermawan
authors.forename: Felix
authors.initials: F
authors.lastname: Solamo
authors.forename: Greg
authors.initials: G
authors.lastname: Flynn
authors.forename: Rosalind
authors.initials: R
authors.lastname: Stenson
authors.forename: Tim
authors.initials: T
authors.lastname: Tressel
authors.forename: Sam
authors.initials: S
authors.lastname: Guhan
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002021:Buffers; D016466:CHO Cells; D002412:Cations; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000302:isolation & purification; D006801:Humans; D006863:Hydrogen-Ion Concentration; D011108:Polymers / Q000737:chemistry; D012117:Resins, Synthetic / Q000737:chemistry; D019346:Sodium Acetate / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000302:isolation & purification; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D002021:Buffers; D002412:Cations; D011108:Polymers; D012117:Resins, Synthetic; D013205:Staphylococcal Protein A; D012965:Sodium Chloride; D019346:Sodium Acetate; D004247:DNA