Over the past several years, monoclonal antibodies have become a key area for the generation of new therapeutics in the biopharmaceutical industry. As a result, development efforts have focused on identifying, characterizing, and reducing impurities to increase product quality and ultimately, the speed to patient. During monoclonal antibody production, full or partial reduction of inter-chain disulfide bonds can occur, leading to domain loss and the formation of low molecular weight (LMW) antibody variants. The presence of LMW can directly impact product yield, production time, and product safety; therefore, controlling LMW remains a key focus in monoclonal antibody development. However, it is often difficult to identify the source of LMW formation during large scale manufacturing, and reproducing the conditions necessary for LMW generation in a lab-scale setting is often a challenging endeavor. In order to execute a reproducible scale-down model of LMW formation, careful coordination amongst all aspects of process development (i.e. upstream, downstream, analytics) is essential to not only induce LMW, but also properly identify and analyze LMW species that form at all stages of production. In this study, we use an end-to-end approach from harvest to purification to create a lab-scale model to investigate LMW formation. We show how process variables affected by sample handling, such as oxygen exposure, sample hold time, and storage temperature, impact the ability to detect and characterize LMW species during normal production. We demonstrate inconsistencies in LMW detection during analytical validation, possibly resulting from strong, noncovalent interactions between antibody domains that mask disulfide reduction. Finally, we propose strategies to reduce LMW formation based on a carefully coordinated process development strategy, resulting in overall better product quality.

End-to-end approach to monitoring and reducing LMW Formation during mAb process development

251st National Meeting (2016)

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