Effect of temperature on the aggregation of an Fc-fusion protein under agitation

Wang, Zekun; Gambe-Gilbuena, Arni; Unzai, Satoru; Uchiyama, Susumu; Torisu, Tetsuo

doi:https://doi.org/10.1016/j.ejps.2025.107320

Effect of temperature on the aggregation of an Fc-fusion protein under agitation

Wang, Zekun; Gambe-Gilbuena, Arni; Unzai, Satoru et al.

European Journal of Pharmaceutical Sciences, 2025, 215, 107320

アクセス数:17件（2026-02-15 21:10 集計）

固定URL: https://hdl.handle.net/11094/103669

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タイトル	Effect of temperature on the aggregation of an Fc-fusion protein under agitation
著者	著者名 Wang, Zekun
	著者名 Gambe-Gilbuena, Arni
	NRID 1000060336592 1000060336592 著者名 Unzai, Satoru
	NRID 1000090335381 1000090335381 ORCID 0000-0002-5181-179X 0000-0002-5181-179X researchmap permalink read0090387 read0090387 Scopus Author ID 35429790100 35429790100 著者名 Uchiyama, Susumu
	NRID 1000010730492 1000010730492 ORCID 0000-0002-8269-7803 0000-0002-8269-7803 researchmap permalink torisutetsuo torisutetsuo Scopus Author ID 57361287600 57361287600 著者名 Torisu, Tetsuo
キーワード等	Agitation stress
	Fc-fusion protein
	Protein aggregation
	Protein stability
	Thermal stress
内容	Wang Z., Gambe-Gilbuena A., Unzai S., et al. Effect of temperature on the aggregation of an Fc-fusion protein under agitation. European Journal of Pharmaceutical Sciences 215, 107320 (2025); https://doi.org/10.1016/j.ejps.2025.107320.
抄録	Mitigating protein aggregation remains a challenge in the development of biopharmaceuticals, and agitation is well known as a stress that can induce protein aggregation. However, the temperature dependence of agitation-induced aggregation is not well understood. In this study, the aggregation of an Fc-fusion protein under agitation stress was investigated at 5, 25, and 40 °C. Soluble and insoluble aggregates were quantified by size-exclusion liquid chromatography and flow imaging microscopy, respectively. Both the aggregation level and the aggregate clusters were temperature dependent. The threshold for the orbital shaking that induced protein aggregation was temperature independent. Although thermal stress at 40 °C increased the number of oligomers, it did not lead to a higher monomer loss in a subsequent agitation at 25 °C. The aggregation induced by agitation stress was suppressed by adding a surfactant or removing the vial headspace, indicating that the aggregation occurred via an interface-mediated pathway. Thus, the observed temperature dependence was attributed to the protein adsorption to the interface and the following interfacial unfolding and aggregation was affected by the temperature. The results emphasized the importance of temperature control during shipping to ensure the quality of drug products. Agitation stability studies at a controlled temperature also provide a deep understanding of the protein aggregation mechanism, which is important for formulation development.
出版者	Elsevier B.V.
収録物名	European Journal of Pharmaceutical Sciences
巻(号)	215
ページ	107320
刊行年月	2025-10-15
言語	英語
ハンドルURL	https://hdl.handle.net/11094/103669
PISSN	09280987
eISSN	18790720
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1016/j.ejps.2025.107320
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	European Journal of Pharmaceutical Sciences. 2025, 215, p. 107320
DC.title	Effect of temperature on the aggregation of an Fc-fusion protein under agitation
DC.creator	Wang, Zekun
	Gambe-Gilbuena, Arni
	Unzai, Satoru
	Uchiyama, Susumu
	Torisu, Tetsuo
DC.publisher	Elsevier B.V.
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2025-10-15
DC.identifier	https://doi.org/10.1016/j.ejps.2025.107320
DC.subject	Agitation stress
	Fc-fusion protein
	Protein aggregation
	Protein stability
	Thermal stress
DC.description	Wang Z., Gambe-Gilbuena A., Unzai S., et al. Effect of temperature on the aggregation of an Fc-fusion protein under agitation. European Journal of Pharmaceutical Sciences 215, 107320 (2025); https://doi.org/10.1016/j.ejps.2025.107320.
DCTERMS.abstract	Mitigating protein aggregation remains a challenge in the development of biopharmaceuticals, and agitation is well known as a stress that can induce protein aggregation. However, the temperature dependence of agitation-induced aggregation is not well understood. In this study, the aggregation of an Fc-fusion protein under agitation stress was investigated at 5, 25, and 40 °C. Soluble and insoluble aggregates were quantified by size-exclusion liquid chromatography and flow imaging microscopy, respectively. Both the aggregation level and the aggregate clusters were temperature dependent. The threshold for the orbital shaking that induced protein aggregation was temperature independent. Although thermal stress at 40 °C increased the number of oligomers, it did not lead to a higher monomer loss in a subsequent agitation at 25 °C. The aggregation induced by agitation stress was suppressed by adding a surfactant or removing the vial headspace, indicating that the aggregation occurred via an interface-mediated pathway. Thus, the observed temperature dependence was attributed to the protein adsorption to the interface and the following interfacial unfolding and aggregation was affected by the temperature. The results emphasized the importance of temperature control during shipping to ensure the quality of drug products. Agitation stability studies at a controlled temperature also provide a deep understanding of the protein aggregation mechanism, which is important for formulation development.
DC.rights	This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	103669
citation_title	Effect of temperature on the aggregation of an Fc-fusion protein under agitation
citation_author	Wang, Zekun
	Gambe-Gilbuena, Arni
	Unzai, Satoru
	Uchiyama, Susumu
	Torisu, Tetsuo
citation_publisher	Elsevier B.V.
citation_language	英語
citation_date	2025-10-15
citation_journal_title	European Journal of Pharmaceutical Sciences
citation_volume	215
citation_firstpage	107320
citation_doi	https://doi.org/10.1016/j.ejps.2025.107320
citation_public_url	https://hdl.handle.net/11094/103669

Effect of temperature on the aggregation of an Fc-fusion protein under agitation

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