Evaluating a Modified High Purity Polysorbate 20 Designed to Reduce the Risk of Free Fatty Acid Particle Formation

Formulation mitigations for particle formation induced by enzymatic hydrolysis of polysorbate 20 in protein-based drug products: insights from a full-factorial longitudinal study

“…Recombinant mAb X was selected for this formulation study because its liquid DP underwent rapid PS20 hydrolysis and generated VPs relatively quickly (i.e., within weeks/months) in real-time stability studies at 5 °C, as shown by our earlier investigations into mAb B (Doshi et al 2021), which is referred to as mAb X herein. The CHO-derived mAb X material was produced using upstream and downstream processes that were representative of typical large-scale mAb manufacturing.…”

“…The CHO-derived mAb X material was produced using upstream and downstream processes that were representative of typical large-scale mAb manufacturing. The VPs isolated by gold filter were previously characterized as FFAs by LC-MS and FTIR microscopy for mAb B (Doshi et al 2021), which is the mAb X studied here. There were no signs of oxidative PS20 degradation by GC-MS (data not shown); furthermore, the mAb X DP formulations studied here included 10 mM l-methionine, which is known to mitigate oxidative degradation of PS20 (Doshi et al 2021).…”

“…The VPs isolated by gold filter were previously characterized as FFAs by LC-MS and FTIR microscopy for mAb B (Doshi et al 2021), which is the mAb X studied here. There were no signs of oxidative PS20 degradation by GC-MS (data not shown); furthermore, the mAb X DP formulations studied here included 10 mM l-methionine, which is known to mitigate oxidative degradation of PS20 (Doshi et al 2021). Therefore, mAb X served as a relevant model for studying the impact of formulation parameters on PS20 hydrolysis and FFA particle formation under representative long-term DP storage conditions (2-8 °C).…”

“…The values for the initial [PS20] and pH were also designed to fall within the ranges previously established for our FFA solubility model (Doshi et al 2015(Doshi et al , 2020. The [mAb] and time points for analyses were selected based on prior VP observations in which the mAb X studied here was referred to as mAb B in the earlier studies (Doshi et al 2021). The outputs selected for this formulation study were generated using stateof-the-art analytical methods that are orthogonal/complementary in nature for studying particle formation and PS20 degradation (Fig.…”

“…In recent years, the biopharmaceutical industry has increasingly reported incidences of and investigations into the root cause of polysorbate degradation and associated particle formation in protein formulations, particularly for recombinant monoclonal antibody (mAb) products (Allmendinger et al 2021;Doshi et al 2021;Graf et al 2021;Gregoritza et al 2022;Roy et al 2021;Zhang et al 2022aZhang et al , 2022bZhang et al , 2021Zhang et al , 2020. We hypothesize that this increasing prevalence of particle observations is the unintended consequence of multiple end-to-end aspects of current industry trends: (1) The reliance on intensified upstream processes to increase cell mass and productivity, which increases the HCP load and product recovery burden for downstream processing (Strube et al 2018;Valente et al 2018).…”