Analysis of viral clearance unit operations for monoclonal antibodies

Abstract

Demonstration of viral clearance is a critical step in assuring the safety of biotechnology products. We generated a viral clearance database that contains product information, unit operation process parameters, and viral clearance data from monoclonal antibody and antibody-related regulatory submissions to FDA. Here we present a broad overview of the database and resulting analyses. We report that the diversity of model viruses tested expands as products transition to late-phase. We also present averages and ranges of viral clearance results by Protein A and ion exchange chromatography steps, low pH chemical inactivation, and virus filtration, focusing on retro- and parvoviruses. For most unit operations, an average log reduction value (LRV, a measure of clearance power) for retrovirus of >4 log(10) were measured. Cases where clearance data fell outside of the anticipated range (i.e., outliers) were rationally explained. Lastly, a historical analysis did not find evidence of any improvement trend in viral clearance over time. The data collectively suggest that many unit operations in general can reliably clear viruses.

Development of adsorptive hybrid filters to enable two-step purification of biologics

“…[37][38][39][40][41] Current regulations require that the manufacturing process demonstrate the ability to clear model viruses to ensure the safety of these cell-linederived products prior to approval, with additional built-in safety for robustness. [42][43][44] AEX chromatography has been frequently applied as a viral clearance step during downstream processing, in addition to impurity clearance of molecular species such as HCP and DNA. 44 This necessitates the claimable viral clearance capability from AHF to eliminate the AEX step.…”

“…[42][43][44] AEX chromatography has been frequently applied as a viral clearance step during downstream processing, in addition to impurity clearance of molecular species such as HCP and DNA. 44 This necessitates the claimable viral clearance capability from AHF to eliminate the AEX step. This strategy is appropriate when the therapeutic protein has a pI higher than the virus particle, as is the case for most IgG mAbs.…”

Viral Clearance by Traditional Operations With Significant Knowledge Gaps (Session II): Cation Exchange Chromatography (CEX) and Detergent Inactivation

“…At the previous viral clearance symposium, insights into the general mechanism of viral clearance by cation exchange chromatography (CEX) were shared by Amgen and Novartis (1). Amgen presented data from multiple monoclonal antibody (mAb) products that consistently revealed Ն4 log 10 murine leukemia virus (MuLV) clearance when run at a pH of 5.0.…”

“…Building on the original data presented two years prior, Amgen had continued its efforts focusing on the strong CEX exchanger Fractogel SO 3 Ϫ and had determined that the dominant mechanism for MuLV clearance was retention of the virus via adsorption instead of inactivation. Though the key findings were presented at the symposium, a more extensive description was eventually published as a peer-reviewed journal article (2).…”

Conference Summary: Gaps, Lessons Learned, and Areas for Improvement

“…A set of action items to address these gaps was drafted, and they were listed in the 2009 conference White Paper (1) and were reiterated in the 2011 meeting invitation letter. Follow-up was left optional and dependent on firms and individuals to take initiative(s).…”