title: Quality by design approach for viral clearance by protein a chromatography.
abstract: Protein A chromatography is widely used as a capture step in monoclonal antibody (mAb) purification processes. Antibodies and Fc fusion proteins can be efficiently purified from the majority of other complex components in harvested cell culture fluid (HCCF). Protein A chromatography is also capable of removing modest levels of viruses and is often validated for viral clearance. Historical data mining of Genentech and FDA/CDER databases systematically evaluated the removal of model viruses by Protein A chromatography. First, we found that for each model virus, removal by Protein A chromatography varies significantly across mAbs, while remains consistent within a specific mAb product, even across the acceptable ranges of the process parameters. In addition, our analysis revealed a correlation between retrovirus and parvovirus removal, with retrovirus data generally possessing a greater clearance factor. Finally, we describe a multivariate approach used to evaluate process parameter impacts on viral clearance, based on the levels of retrovirus-like particles (RVLP) present among process characterization study samples. It was shown that RVLP removal by Protein A is robust, that is, parameter effects were not observed across the ranges tested. Robustness of RVLP removal by Protein A also correlates with that for other model viruses such as X-MuLV, MMV, and SV40. The data supports that evaluating RVLP removal using process characterization study samples can establish multivariate acceptable ranges for virus removal by the protein A step for QbD. By measuring RVLP instead of a model retrovirus, it may alleviate some of the technical and economic challenges associated with performing large, design-of-experiment (DoE)-type virus spiking studies. This approach could also serve to provide useful insight when designing strategies to ensure viral safety in the manufacturing of a biopharmaceutical product.
authors.affiliation: Process Virology, Purification Development, MS 10, Genentech, Inc., 1 DNA Way, South San Francisco, California, 04080.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: George R
authors.initials: GR
authors.lastname: Miesegaes
authors.forename: Michael
authors.initials: M
authors.lastname: Lee
authors.forename: Daniel
authors.initials: D
authors.lastname: Coleman
authors.forename: Bin
authors.initials: B
authors.lastname: Yang
authors.forename: Melody
authors.initials: M
authors.lastname: Trexler-Schmidt
authors.forename: Lenore
authors.initials: L
authors.lastname: Norling
authors.forename: Philip
authors.initials: P
authors.lastname: Lester
authors.forename: Kurt A
authors.initials: KA
authors.lastname: Brorson
authors.forename: Qi
authors.initials: Q
authors.lastname: Chen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001688:Biological Products / Q000302:isolation & purification / Q000592:standards; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D015999:Multivariate Analysis; D015203:Reproducibility of Results; D012190:Retroviridae / Q000302:isolation & purification*; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D001688:Biological Products; D013205:Staphylococcal Protein A
keywords: MMV; QbD; SV40; X-MuLV; protein A chromatography; retrovirus-like particles; risk ranking; virus removal