Factors Influencing Polysorbate's Sensitivity Against Enzymatic Hydrolysis and Oxidative Degradation

Kranz, Wendelin; Wuchner, Klaus; Corradini, Eleonora; Berger, Michelle; Hawe, Andrea

doi:10.1016/j.xphs.2019.01.006

Cited by 65 publications

(92 citation statements)

References 30 publications

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“…PS80 was reported neither to have an effect on the thermostability of IgGs 37 nor to inhibit oxidation in IgG formulations. 19 Therefore, the inhibition of particle formation seen in formulations containing PS80 suggests interface-mediated aggregation as the predominant factor causing aggregation in dropped vials. Particle formation in formulations exposed to free-fall stress was suppressed in presence of PS80, and even at low PS80 concentrations (below CMC), solely a very small increase of micron-sized particles could be observed.…”

Section: Discussionmentioning

confidence: 99%

“…Functionality of both PS80 grades after long-term storage was not investigated but should be considered given the recently described differences in stability of MC-PS80 and ChP-PS80. 19 Further studies would also be necessary to evaluate if the "harsh" syringe pump stress conditions correlate with current Fill & Finish manufacturing conditions. Our study points out that it is important to monitor PS during drug product formulation development and stability testing of therapeutic proteins.…”

Section: Discussionmentioning

confidence: 99%

“…Characterization and quantification of PS80 and its degradation products was performed by using the method developed by Kranz et al 19 A Dionex Ultimate 3000 HPLC system hyphenated with a Corona Veo CAD (Thermo Scientific) was used. Separation was achieved by using a BEH C18 column, 50 Â 2.1 mm (Waters) at 60 C. A 6-point calibration curve was generated for quantification of PS80 supplemented to the IgG formulations that were investigated in this study.…”

Section: High Performance Liquid Chromatography Coupled With a Chargementioning

confidence: 99%

“…Nonetheless, commercial PS products consist of a mixture of monoesters and polyesters of POE sorbitans alongside substantial amounts of POE, sorbitan POE, and isosorbide POE fatty acid esters, originating from the synthesis. 18 Significant variations in the chemical composition of PS can be present, not only between suppliers 19 but also between different lots of a single supplier. 20 In addition, PS is known to be susceptible to oxidative and hydrolytic (mainly enzymatic under pharmaceutical relevant conditions 21 ) degradation, resulting in the formation of aldehydes, epoxides, free fatty acids (FFAs), and other impurities, which adds on to their heterogeneity.…”

Section: Introductionmentioning

confidence: 99%

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What Makes Polysorbate Functional? Impact of Polysorbate 80 Grade and Quality on IgG Stability During Mechanical Stress

Grabarek

Bozic²,

Rousel

et al. 2020

Journal of Pharmaceutical Sciences

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Polysorbate 80 (PS80) is a commonly used surfactant in therapeutic protein formulations to mitigate adsorption and interface-induced protein aggregation. Several PS80 grades and qualities are available on the market for parenteral application. The role of PS80 grade on protein stability remains debatable, and the impact of (partially) degraded PS on protein aggregation is not yet well understood. In our study, a monoclonal antibody (IgG) was subjected to 3 different mechanical stress conditions in the presence of multicompendial (MC) and Chinese pharmacopeia (ChP) grade PS80. Furthermore, IgG formulations were spiked with (partly) hydrolyzed PS80 to investigate the effect of PS80 degradants on protein stability. PS80 functionality was assessed by measuring the extent of protein aggregation and particle formation induced during mechanical stress by using size-exclusion chromatography, dynamic light scattering, backgrounded membrane imaging, and flow imaging microscopy. No distinguishable differences in PS80 functionality between MC and ChP grade were observed in the 3 stress tests. However, with increasing degree of PS80 hydrolysis, higher counts of subvisible particles were measured after stress. Furthermore, higher levels of PS80 degradants at a constant PS80 concentration may destabilize the IgG. In conclusion, MC and ChP grade PS80 are equally protective, but PS80 degradants compromise IgG stability.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: High Performance Liquid Chromatography Coupled With a Chargementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

What Makes Polysorbate Functional? Impact of Polysorbate 80 Grade and Quality on IgG Stability During Mechanical Stress

Grabarek

Bozic²,

Rousel

et al. 2020

Journal of Pharmaceutical Sciences

Self Cite

View full text Add to dashboard Cite

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“…164 In fact, only a few ppb of oxidative metals can be enough to degrade the protein as well as excipients (e.g., polysorbates) in the formulation that can be sensitive to this type of oxidative stress. 165…”

Section: Leachables and Extractablesmentioning

confidence: 99%

Stress Factors in mAb Drug Substance Production Processes: Critical Assessment of Impact on Product Quality and Control Strategy

Das

Narhi

Sreedhara³

et al. 2020

Journal of Pharmaceutical Sciences

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The success of biotherapeutic development heavily relies on establishing robust production platforms. During the manufacturing process, the protein is exposed to multiple stress conditions that can result in physical and chemical modifications. The modified proteins may raise safety and quality concerns depending on the nature of the modification. Therefore, the protein modifications potentially resulting from various process steps need to be characterized and controlled. This commentary brings together expertise and knowledge from biopharmaceutical scientists and discusses the various manufacturing process steps that could adversely impact the quality of drug substance (DS). The major process steps discussed here are commonly used in mAb production using mammalian cells. These include production cell culture, harvest, antibody capture by protein A, virus inactivation, polishing by ion-exchange chromatography, virus filtration, ultrafiltration-diafiltration, compounding followed by release testing, transportation and storage of final DS. Several of these process steps are relevant to protein DS production in general. The authors attempt to critically assess the level of risk in each of the DS processing steps, discuss strategies to control or mitigate protein modification in these steps, and recommend mitigation approaches including guidance on development studies that mimic the stress induced by the unit operations.

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Detailed profiling of polysorbate 80 oxidative degradation products and hydrolysates using liquid chromatography–tandem mass spectrometry analysis

Konya,

Ochiai,

Fujiwara

et al. 2024

Rapid Comm Mass Spectrometry

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RationalePolysorbate 80 (PS80) is an amphipathic, nonionic surfactant that is commonly used to stabilize proteins in biopharmaceutical formulations. PS80 undergoes oxidative and/or enzymatic degradation. However, because PS80 is a complex mixture consisting of many constituents, comprehensive evaluations of its oxidative degradation products are difficult and insufficient.MethodsOur previously reported comprehensive liquid chromatography–tandem mass spectrometry (LC/MS/MS)‐based method for PS80 effectively provides an overall profile of PS80 components under simple LC conditions. In this study, we attempted to shorten the analysis time. Furthermore, PS80 was oxidatively degraded in a solution containing histidine and iron, and the oxidative degradation products were evaluated using a modified LC/MS/MS method. In addition, enzymatically hydrolyzed PS80 samples were analyzed.ResultsWe succeeded in shortening the analysis time from 70 to 20 min while maintaining the resolution of the PS80 components of the same selected reaction monitoring transition. Both the previously reported oxidative degradation products and the newly discovered products were successfully detected, and their composition ratios and changes over time were observed. Changes in the hydrolysates over time are shown in the analysis of the hydrolyzed PS80 samples.ConclusionsThis study clearly showed the presence of changes in PS80 oxidative and/or enzymatic degradation products, including those previously unreported. These results demonstrate that a detailed profiling of PS80 degradation products can be performed using LC/MS/MS, which is less expensive and more generally adopted than high‐resolution MS.

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Factors Influencing Polysorbate's Sensitivity Against Enzymatic Hydrolysis and Oxidative Degradation

Cited by 65 publications

References 30 publications

What Makes Polysorbate Functional? Impact of Polysorbate 80 Grade and Quality on IgG Stability During Mechanical Stress

What Makes Polysorbate Functional? Impact of Polysorbate 80 Grade and Quality on IgG Stability During Mechanical Stress

Stress Factors in mAb Drug Substance Production Processes: Critical Assessment of Impact on Product Quality and Control Strategy

Detailed profiling of polysorbate 80 oxidative degradation products and hydrolysates using liquid chromatography–tandem mass spectrometry analysis

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