Formation of protein sub-visible particles during vacuum degassing of etanercept solutions

Wang, Haibin; Zheng, Hong-Jian; Wang, Zhao; Bai, Hua; Carpenter, John F.; Chen, Shuqing; Fang, Wei-Jie

doi:10.1016/j.ijbiomac.2014.02.001

Cited by 8 publications

(5 citation statements)

References 21 publications

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“…Similar discrepancy has been reported previously in several other cases. 27,28 Traditional techniques such as SE-HPLC and particle-analyzing techniques are detecting somewhat related but different quality attributes, especially for high concentration formulations (i.e., >10 mg/mL for most mAb formulations), where the formation of substantial sub-visible particles may only account for tiny amount of mass. However, for low concentration protein formulations, formation of substantial sub-visible particles could be correlated to monomer loss as determined by traditional methods such as SEC.…”

Section: Discussionmentioning

confidence: 99%

Protein Sub-Visible Particle and Free Radical formation of a Freeze-Dried Monoclonal Antibody Formulation During Dropping

Fang

Liu

Zheng

et al. 2021

Journal of Pharmaceutical Sciences

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Dropping during shipping and handling of liquid biopharmaceutical formulations has long been known to cause protein degradation and aggregation. On the other hand, accidental dropping of freeze-dried protein formulations is generally considered not a major issue for biopharmaceutical quality. Reports of stability and especially the underling degradation mechanism(s) during shipping and handling of freeze-dried protein formulations were rarely seen in literature. In this manuscript, we report an interesting phenomenon in which repeated dropping of freeze-dried monoclonal antibody X (mAb-X) formulation powder resulted in significant protein sub-visible particles (SbVPs) in the reconstituted liquid as determined by the sensitive particle analyzing technique micro-flow imaging (MFI). Free radicals were observed after repeated dropping by electron paramagnetic resonance (EPR). Formation of SbVPs could be partially inhibited by the free radical scavengers methionine and 3-carbamoyl-2,2,5,5tetramethyl-1-pyrrolidin-yloxy free radical (CTPO). The amount of free radicals and SbVPs was correlated to the sample temperature during dropping. Therefore we propose that the high temperature formed during dropping was probably the root cause for protein aggregation and free radical formation, which could further cause protein aggregation. Our observations suggest that similar to liquid protein formulations, dropping of freeze-dried protein formulations should also be avoided or mitigated.

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Section: Discussionmentioning

confidence: 99%

Protein Sub-Visible Particle and Free Radical formation of a Freeze-Dried Monoclonal Antibody Formulation During Dropping

Fang

Liu

Zheng

et al. 2021

Journal of Pharmaceutical Sciences

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“…Degassing can be used to avoid air bubbles, which can occur, for example, upon heating a cooled sample or during reconstitution of lyophilized product. Generally, vacuum degassing is sufficiently gentle to remove air bubbles without destroying the protein particles; however, it has been shown that degassing can induce changes in the aggregation profile of some samples . Alternatively, air bubbles can be avoided by letting the product sit for defined periods of time.…”

Section: Testing Considerationsmentioning

confidence: 99%

“…Generally, vacuum degassing is sufficiently gentle to remove air bubbles without destroying the protein particles; however, it has been shown that degassing can induce changes in the aggregation profile of some samples. 60 Alternatively, air bubbles can be avoided by letting the product sit for defined periods of time. Sample degassing should be carefully assessed on a product-by-product basis.…”

Section: Sample Handlingmentioning

confidence: 99%

Subvisible (2-100 μm) Particle Analysis During Biotherapeutic Drug Product Development: Part 1, Considerations and Strategy

Narhi

Corvari

Ripple³

et al. 2015

Journal of Pharmaceutical Sciences

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“…These particles could be the potentially most immunogenic class of protein aggregates and may act as nuclei ( 25 ) and cause formation of larger particles over time ( 5 , 26 ). In order to detect and assign aggregates/particles to the categories mentioned above we have employed Micro Flow Imaging (MFI) and Dynamic Light Scattering (DLS) ( 27 – 29 ). MFI is used for detecting particles in size ranges from 2 to 300 μm and DLS to detect and characterize soluble aggregates on a length scale of ca.…”

Section: Introductionmentioning

confidence: 99%

Impact of Buffer, Protein Concentration and Sucrose Addition on the Aggregation and Particle Formation during Freezing and Thawing

Hauptmann

Podgoršek²,

Kuzman³

et al. 2018

Pharm Res

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PurposeThis study addresses the effect of freezing and thawing on a therapeutic monoclonal antibody (mAb) solution and the corresponding buffer formulation. Particle formation, crystallization behaviour, morphology changes and cryo-concentration effects were studied after varying the freezing and thawing rates, buffer formulation and protein concentration. The impact of undergoing multiple freeze/thaw (FT)-cycles at controlled and uncontrolled temperature rates on mAb solutions was investigated in terms of particle formation.MethodsPhysicochemical characteristics were analysed by Differential Scanning Calorimetry whereas morphology changes are visualized by cryomicroscopy measurements. Micro Flow Imaging, Archimedes and Dynamic Light Scattering were used to investigate particle formation.ResultsData retrieved in the present study emphasizes the damage caused by multiple FT-cyles and the need for sucrose as a cryoprotectant preventing cold-crystallization specifically at high protein concentrations. Low protein concentrations cause an increase of micron particle formation. Low freezing rates lead to a decreased particle number with increased particle diameter.ConclusionThe overall goal of this research is to gain a better understanding of the freezing and thawing behaviour of mAb solutions with the ultimate aim to optimize this process step by reducing the unwanted particle formation, which also includes protein aggregates.

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Formation of protein sub-visible particles during vacuum degassing of etanercept solutions

Cited by 8 publications

References 21 publications

Protein Sub-Visible Particle and Free Radical formation of a Freeze-Dried Monoclonal Antibody Formulation During Dropping

Protein Sub-Visible Particle and Free Radical formation of a Freeze-Dried Monoclonal Antibody Formulation During Dropping

Subvisible (2-100 μm) Particle Analysis During Biotherapeutic Drug Product Development: Part 1, Considerations and Strategy

Impact of Buffer, Protein Concentration and Sucrose Addition on the Aggregation and Particle Formation during Freezing and Thawing

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