title: Development of a modular virus clearance package for anion exchange chromatography operated in weak partitioning mode.
abstract: Anion exchange chromatography (AEX) operated under weak partitioning mode has been proven to be a powerful polishing step as well as a robust viral clearance step in Pfizer's monoclonal antibody (mAb) platform purification process. A multivariate design of experiment (DoE) study was conducted to understand the impact of operating parameters and feedstream impurity levels on viral clearance by weak partitioning mode AEX. Bacteriophage was used initially as a surrogate for neutral and acidic isoelectric point mammalian viruses (e.g., retrovirus and parvovirus). Five different mAbs were used in the evaluation of process parameters such as load challenge (both product and impurities), load pH, load conductivity, and contact time (bed height and flow-rate). The operating ranges obtained from phage clearance studies and Pfizer's historical data were used to define an appropriate operating range for a subsequent clearance study with model retrovirus and parvovirus. Both phage and virus clearance evaluations included feedstreams containing different levels of impurities such as high molecular mass species (HMMS), host cell proteins (HCPs), and host cell DNA. For all the conditions tested, over 5 log10 of clearance for both retrovirus and parvovirus was achieved. The results demonstrated that weak partitioning mode AEX chromatography is a robust step for viral clearance and has the potential to be included as part of the modular viral clearance approach.
authors.affiliation: Pfizer Inc., Purification Process Development, Andover, MA, 01810.
authors.forename: Timothy
authors.initials: T
authors.lastname: Iskra
authors.affiliation: Pfizer Inc., Purification Process Development, Andover, MA, 01810.
authors.forename: Ashley
authors.initials: A
authors.lastname: Sacramo
authors.affiliation: Pfizer Inc., Purification Process Development, Andover, MA, 01810.
authors.forename: Chris
authors.initials: C
authors.lastname: Gallo
authors.affiliation: Pfizer Inc., Purification Process Development, Andover, MA, 01810.
authors.forename: Ranga
authors.initials: R
authors.lastname: Godavarti
authors.affiliation: Pfizer Inc., Purification Process Development, Chesterfield, MO, 63017.
authors.forename: Shuang
authors.initials: S
authors.lastname: Chen
authors.affiliation: Office of Biotechnology Products, CDER/FDA, 10903 New Hampshire Ave., Silver Spring, MD, 20903.
authors.forename: Scott
authors.initials: S
authors.lastname: Lute
authors.affiliation: Office of Biotechnology Products, CDER/FDA, 10903 New Hampshire Ave., Silver Spring, MD, 20903.
authors.forename: Kurt
authors.initials: K
authors.lastname: Brorson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001435:Bacteriophages / Q000302:isolation & purification; D016466:CHO Cells; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003412:Cricetulus; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D017992:Parvovirus / Q000302:isolation & purification*; D012190:Retroviridae / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal
keywords: anion-exchange chromatography; monoclonal antibody; purification; viral clearance