Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1

Hitomi, Natsuki; Hoshi, Satowa; Kaneko, Mika K.; Kato, Ryuichi; Iwasaki, Kenji; Takagi, Junichi; Kato, Yukinari; Harada-Hikita, Ayaka; Arimori, Takao

doi:https://doi.org/10.1016/j.jmb.2025.169574

Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1

Hitomi, Natsuki; Hoshi, Satowa; Kaneko, Mika K. et al.

Journal of Molecular Biology, 2026, 438(2), 169574

アクセス数:45件（2026-02-01 06:34 集計）

固定URL: https://hdl.handle.net/11094/103579

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タイトル	Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1
著者	著者名 Hitomi, Natsuki
	著者名 Hoshi, Satowa
	NRID 1000000323163 1000000323163 著者名 Kaneko, Mika K.
	NRID 1000050240833 1000050240833 著者名 Kato, Ryuichi
	NRID 1000020342751 1000020342751 著者名 Iwasaki, Kenji
	NRID 1000090212000 1000090212000 researchmap permalink read0105692 read0105692 Scopus Author ID 7102173538 7102173538 著者名 Takagi, Junichi
	NRID 1000000571811 1000000571811 著者名 Kato, Yukinari
	NRID 1000000800476 1000000800476 著者名 Harada-Hikita, Ayaka
	NRID 1000080582064 1000080582064 ORCID 0000-0002-6063-5572 0000-0002-6063-5572 researchmap permalink t_arimori t_arimori Scopus Author ID 35336011300 35336011300 著者名 Arimori, Takao
キーワード等	antibody
	isothermal titration calorimetry
	polyhistidine tag
	surface plasmon resonance
	X-ray crystallography
内容	Hitomi N., Hoshi S., Kaneko M.K., et al. Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1. Journal of Molecular Biology 438, 169574 (2026); https://doi.org/10.1016/j.jmb.2025.169574.
抄録	The polyhistidine tag (His-tag) is one of the most widely used peptide tags for the purification of recombinant proteins, owing to its compatibility with immobilized metal affinity chromatography. While numerous anti-His-tag antibodies are commercially available, their quantitative affinity data and structural insights are limited. Here, we present a detailed physicochemical and structural characterization of a novel anti-His-tag antibody, HisMab-1. Isothermal titration calorimetry showed that the Fab fragment of HisMab-1 binds to a hexahistidine peptide in an enthalpy-driven manner, with a dissociation constant (KD) of ∼30 nM at a neutral pH. The crystal structure of the HisMab-1–hexahistidine peptide complex at 2.39-Å resolution revealed that HisMab-1 primarily recognizes the first, second, fourth, and fifth histidine residues of the peptide through multiple interactions, including hydrogen bonding and π–π stacking, which collectively contribute to the high specificity of the antibody. Notably, HisMab-1 also binds to a His-tag embedded within a conformationally constrained β-hairpin loop without reducing affinity, highlighting its structural adaptability. These findings establish HisMab-1 as a high-affinity, high-specificity, structurally validated anti-His-tag antibody with broad potential in diverse protein engineering and structural biology applications.
出版者	Academic Press
収録物名	Journal of Molecular Biology
巻(号)	438(2)
ページ	169574
刊行年月	2026-01-15
言語	英語
ハンドルURL	https://hdl.handle.net/11094/103579
PISSN	00222836
eISSN	10898638
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.1016/j.jmb.2025.169574
アクセス権	open access
権利情報	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 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	Journal of Molecular Biology. 2026, 438(2), p. 169574
DC.title	Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1
DC.creator	Hitomi, Natsuki
	Hoshi, Satowa
	Kaneko, Mika K.
	Kato, Ryuichi
	Iwasaki, Kenji
	Takagi, Junichi
	Kato, Yukinari
	Harada-Hikita, Ayaka
	Arimori, Takao
DC.publisher	Academic Press
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2026-01-15
DC.identifier	https://doi.org/10.1016/j.jmb.2025.169574
DC.subject	antibody
	isothermal titration calorimetry
	polyhistidine tag
	surface plasmon resonance
	X-ray crystallography
DC.description	Hitomi N., Hoshi S., Kaneko M.K., et al. Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1. Journal of Molecular Biology 438, 169574 (2026); https://doi.org/10.1016/j.jmb.2025.169574.
DCTERMS.abstract	The polyhistidine tag (His-tag) is one of the most widely used peptide tags for the purification of recombinant proteins, owing to its compatibility with immobilized metal affinity chromatography. While numerous anti-His-tag antibodies are commercially available, their quantitative affinity data and structural insights are limited. Here, we present a detailed physicochemical and structural characterization of a novel anti-His-tag antibody, HisMab-1. Isothermal titration calorimetry showed that the Fab fragment of HisMab-1 binds to a hexahistidine peptide in an enthalpy-driven manner, with a dissociation constant (KD) of ∼30 nM at a neutral pH. The crystal structure of the HisMab-1–hexahistidine peptide complex at 2.39-Å resolution revealed that HisMab-1 primarily recognizes the first, second, fourth, and fifth histidine residues of the peptide through multiple interactions, including hydrogen bonding and π–π stacking, which collectively contribute to the high specificity of the antibody. Notably, HisMab-1 also binds to a His-tag embedded within a conformationally constrained β-hairpin loop without reducing affinity, highlighting its structural adaptability. These findings establish HisMab-1 as a high-affinity, high-specificity, structurally validated anti-His-tag antibody with broad potential in diverse protein engineering and structural biology applications.
DC.rights	This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
DC.format	103579
citation_title	Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1
citation_author	Hitomi, Natsuki
	Hoshi, Satowa
	Kaneko, Mika K.
	Kato, Ryuichi
	Iwasaki, Kenji
	Takagi, Junichi
	Kato, Yukinari
	Harada-Hikita, Ayaka
	Arimori, Takao
citation_publisher	Academic Press
citation_language	英語
citation_date	2026-01-15
citation_journal_title	Journal of Molecular Biology
citation_volume	438
citation_issue	2
citation_firstpage	169574
citation_doi	https://doi.org/10.1016/j.jmb.2025.169574
citation_public_url	https://hdl.handle.net/11094/103579

Functional and Structural Characterization of a Novel Anti-His-tag Antibody, HisMab-1

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