Metal-Induced Fatty Acid Particle Formation Resulting from Hydrolytic Polysorbate Degradation

Gregoritza, Kathrin; Cai, Si Kai; Siketanc, Matej; Woehr, Anne; Lebouc, Vanessa; Kishore, Ravuri S. K.; Nicoulin, Victor; Bleher, Stefan; Allmendinger, Andrea

doi:10.1016/j.xphs.2021.09.044

Cited by 9 publications

(8 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The presence of aluminum ions was reported to induce fatty acid particles in the case free fatty acids are generated e.g. by enzymatic degradation [ 71 ], and an impact on oxidation can be discussed. It was also reported to have a propensity to be involved in the oxidation of oils [ 72 ].…”

Section: Discussionmentioning

confidence: 99%

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Kozuch,

Weber,

Buske

et al. 2023

Pharmaceutics

View full text Add to dashboard Cite

The surfactants polysorbate 20 (PS20) and polysorbate 80 (PS80) are utilized to stabilize protein drugs. However, concerns have been raised regarding the degradation of PSs in biologics and the potential impact on product quality. Oxidation has been identified as a prevalent degradation mechanism under pharmaceutically relevant conditions. So far, a systematic stability comparison of both PSs under pharmaceutically relevant conditions has not been conducted and little is known about the dependence of oxidation on PS concentration. Here, we conducted a comparative stability study to investigate (i) the different oxidative degradation propensities between PS20 and PS80 and (ii) the impact of PS concentration on oxidative degradation. PS20 and PS80 in concentrations ranging from 0.1 mg⋅mL−1 to raw material were stored at 5, 25, and 40 °C for 48 weeks in acetate buffer pH 5.5 and water, respectively. We observed a temperature-dependent oxidative degradation of the PSs with strong (40 °C), moderate (25 °C), and weak/no degradation (5 °C). Especially at elevated temperatures such as 40 °C, fast oxidative PS degradation processes were detected. In this case study, a stronger degradation and earlier onset of oxidation was observed for PS80 in comparison to PS20, detected via the fluorescence micelle assay. Additionally, degradation was found to be strongly dependent on PS concentration, with significantly less oxidative processes at higher PS concentrations. Iron impurities, oxygen in the vial headspaces, and the pH values of the formulations were identified as the main contributing factors to accelerate PS oxidation.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Kozuch,

Weber,

Buske

et al. 2023

Pharmaceutics

View full text Add to dashboard Cite

show abstract

“…However, they do support a positive trend between the rise in SVP counts and the onset of VPs for mAb X DP. Given the complexity of the particle nucleation phenomenon (Allmendinger et al 2021;Gregoritza et al 2022;Roy et al 2021;Saggu et al 2021), and the relatively early onset of particles in the mAb X formulations, the overall trends between SVPs and VPs observed here may not be generalizable.…”

Section: Correlations Between Visible and Subvisible Particlesmentioning

confidence: 86%

“…Based on the findings here and considerations elsewhere on the application of SVPs in the 2-100-µm size range (Carpenter et al 2009;Corvari et al 2015), the data from the smallest SVP size category (≥ 2 µm) were further analyzed for the impact of formulation parameters. A low SVP count threshold (< 350 particles/mL) was applied based on in-house experience to approximate the onset of the rise in SVP counts above background noise (Gregoritza et al 2022). Lower [mAb] and higher initial [PS20] extended the storage duration with low SVP counts (Fig.…”

Section: Subvisible Particles (Svps)mentioning

confidence: 99%

“…In other in-house studies, VPs that formed earliest were comprised primarily of SA and PA, whereas VPs that formed later included MA with SA and PA in its composition (Saggu et al 2021). Another limitation of the FFA solubility model is its lack of coverage for nucleation factors, which have been shown to trigger the onset of VPs below predicted FFA solubility limits (Allmendinger et al 2021;Gregoritza et al 2022). In spite of these limitations, the FFA solubility model increased our understanding of the observed impact of the formulation parameters on VP outcomes in the ranges tested in this longitudinal DoE study.…”

Section: Comparison Of Model Predictions With Experimental Observationsmentioning

confidence: 99%

“…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).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Yuk¹,

Koulis²,

Doshi³

et al. 2022

AAPS Open

Self Cite

View full text Add to dashboard Cite

Hydrolytic degradation of the polysorbate 20 (PS20) surfactant in protein-based liquid formulations releases free fatty acids (FFAs), which can accumulate to form particles in drug products during real-time (long-term) storage. To identify formulation conditions that mitigate the risk of particle formation, we conducted a longitudinal study using purified recombinant monoclonal antibody (mAb) formulated in 24 conditions. In this real-time stability study at 5 °C, three key formulation parameters—mAb concentration, initial PS20 concentration, and pH—were varied across representative ranges in a full-factorial design. A longitudinal regression analysis was used to evaluate the effects of these parameters and their interactions on PS20 degradation (via measurements of PS20, FFAs, and PS20 ester distribution) and on particle formation (via visible particle observations and subvisible particle counts). The time-dependent onset of visible particles trended with the rise in subvisible particle counts and FFA levels and fall in PS20 concentration. In the ranges studied here, lower mAb concentration and higher initial PS20 concentration delayed the onset of particles, whereas pH had a negligible effect. These observations were consistent with the general trends predicted by our previously published FFA solubility model. Taken together, these findings highlight the complex relationships between formulation parameters, PS20 degradation, and particle formation.

show abstract

Stress-mediated polysorbate 20 degradation and its potential impact on therapeutic proteins

Aryal,

Lehtimaki,

Rao

2024

Pharm Res

View full text Add to dashboard Cite

Purpose Polysorbates are the most commonly used surfactants in formulations to stabilize therapeutic proteins against interfacial stresses. Polysorbates can undergo oxidative or enzyme-mediated hydrolytic degradation to produce free fatty acids (FFAs) and subvisible particles in formulations. To determine which product related variables contribute to PS20 degradation, we investigated the effects of storage temperature, formulation, pH, presence of hydrolytic enzymes, and specific fatty acid composition on different grades of PS20 in relation to their PS20 degradation profile and consequently the quality of protein drug products. Methods Bevacizumab and T-DM1 were reformulated in the freshly prepared therapeutic protein formulations containing either compendial PS20 or non-compendial PS20 with high % lauric acid and spiked with exogenous esterase or lipase. The release of FFAs and formation of particles were monitored at 4°C and 37°C. Protein quality was assessed for secondary structures, purity, and biological activity. Results Hydrolytic release of FFAs and formation of subvisible particles were found to be dependent on grades of PS20, types of enzymes used, incubation temperature, and pH. Esterase- or lipase-mediated degradation of PS20 and formation of subvisible particles in drug formulation showed no significant impact on the biological activity and stability of therapeutic proteins against degradation or aggregation. Conclusions Our study suggests that degradation of PS20 and formation of FFA particles depend on the fatty acid composition of PS20, types of hydrolytic enzymes, pH, and temperature. The presence of FFA subvisible particles showed no significant impact on the purity and biological activity of the therapeutic proteins under the tested conditions.

show abstract

Metal-Induced Fatty Acid Particle Formation Resulting from Hydrolytic Polysorbate Degradation

Cited by 9 publications

References 36 publications

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

Comparative Stability Study of Polysorbate 20 and Polysorbate 80 Related to Oxidative Degradation

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

Stress-mediated polysorbate 20 degradation and its potential impact on therapeutic proteins

Contact Info

Product

Resources

About