Antibody Reduction in Bioprocessing: Analytical Detection and Steps for Mitigation
Antibody reduction is the spontaneous degradation of inter-chain disulfide bonds resulting in a build-up of partial related substances of an IgG in bioprocess streams. This can lead to decreased product yield and altered batch quality and efficacy profile. Process factors known to favor antibody reduction are the antibody subclass, high shear force during harvest, low dissolved oxygen during harvest and storage conditions (pH, temperature, time) of harvest material. This webinar will highlight a case study of a new monoclonal antibody undergoing process optimization where antibody reduction was encountered showing partial related substances present at 38 percent by area.
The webinar will focus on the analytical detection and characterization technologies employed and the in-process control data used to understand the potential sources of degradation in antibody reduction. A capillary electrophoresis system was used to first detect the related substances present in the demonstration run drug substance using non-reduced CE-SDS analysis. The results were verified by retesting and by altering sample preparation conditions to eliminate method artifacts as a potential source of the species. Full analytical panel testing was performed on in-process retains to identify key process factors for the reduction and to better understand the nature and impact of the related substances in the product batch. Additional testing included SDS-PAGE, SE-HPLC, LC-MS, ELISA potency, icIEF and peptide map by RP-HPLC.
The process team used the analytical data to identify the potential antibody reduction sources with their respective mitigation strategies. Key mitigation steps were divided into prevention and rescue of reduced species. Process mitigations were focused on the harvest conditions and by adding a redox wash step to the Protein A capture column prior to elution. The process procedure was updated to incorporate the critical changes and a new demonstration run was performed to study the impact on the presence of related substances. The optimized process drug substance showed expected yield and an effective removal of the related substances.
Speaker
Aaron Martin has 15 years of experience in the biotech/pharmaceutical industry. He currently leads the Analytical Development group within Cytovance Biologics R&D Services. In this role, he oversees the transfer, development and phase-appropriate qualification of analytical methods for new API programs, with a focus on ensuring that analytical methods and method qualifications are fit-for-purpose for their intended use.
Aaron is a subject-matter expert in biomolecular binding assays, in particular with SPR and BLI technologies. Prior to working at Cytovance, he held positions at ForteBio and SensiQ Technologies where he gained extensive experience with method development and contract research for scientists in academia and the pharmaceutical industry.
Join Aaron Martin, Associate Director of Analytical Development, for the live webinar on Tuesday, May 23, 2023 | 12pm EDT (US) | 11am CT (US). For more information, or to register for this event, visit Antibody Reduction in Bioprocessing: Analytical Detection and Steps for Mitigation.