Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope

Abstract Antibodies have been widely used for cancer therapy owing to their ability to distinguish cancer cells by recognizing cancer-specific antigens. Epidermal growth factor receptor (EGFR) is a promising target for the cancer therapeutics, against which several antibody clones have been develope...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Koki Makabe, Takeshi Yokoyama, Shiro Uehara, Tomomi Uchikubo-Kamo, Mikako Shirouzu, Kouki Kimura, Kouhei Tsumoto, Ryutaro Asano, Yoshikazu Tanaka, Izumi Kumagai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/051aab10792249469899ed21bbfad1a2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:051aab10792249469899ed21bbfad1a2
record_format	dspace
spelling	oai:doaj.org-article:051aab10792249469899ed21bbfad1a22021-12-02T11:35:52ZAnti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope10.1038/s41598-021-84171-32045-2322https://doaj.org/article/051aab10792249469899ed21bbfad1a22021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-84171-3https://doaj.org/toc/2045-2322Abstract Antibodies have been widely used for cancer therapy owing to their ability to distinguish cancer cells by recognizing cancer-specific antigens. Epidermal growth factor receptor (EGFR) is a promising target for the cancer therapeutics, against which several antibody clones have been developed and brought into therapeutic use. Another antibody clone, 528, is an antagonistic anti-EGFR antibody, which has been the focus of our antibody engineering studies to develop cancer drugs. In this study, we explored the interaction of 528 with the extracellular region of EGFR (sEGFR) via binding analyses and structural studies. Dot blotting experiments with heat treated sEGFR and surface plasmon resonance binding experiments revealed that 528 recognizes the tertiary structure of sEGFR and exhibits competitive binding to sEGFR with EGF and cetuximab. Single particle analysis of the sEGFR–528 Fab complex via electron microscopy clearly showed the binding of 528 to domain III of sEGFR, the domain to which EGF and cetuximab bind, explaining its antagonistic activity. Comparison between the two-dimensional class average and the cetuximab/sEGFR crystal structure revealed that 528 binds to a site that is shifted from, rather than identical to, the cetuximab epitope, and may exclude known drug-resistant EGFR mutations.Koki MakabeTakeshi YokoyamaShiro UeharaTomomi Uchikubo-KamoMikako ShirouzuKouki KimuraKouhei TsumotoRyutaro AsanoYoshikazu TanakaIzumi KumagaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-6 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Koki Makabe Takeshi Yokoyama Shiro Uehara Tomomi Uchikubo-Kamo Mikako Shirouzu Kouki Kimura Kouhei Tsumoto Ryutaro Asano Yoshikazu Tanaka Izumi Kumagai Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
description	Abstract Antibodies have been widely used for cancer therapy owing to their ability to distinguish cancer cells by recognizing cancer-specific antigens. Epidermal growth factor receptor (EGFR) is a promising target for the cancer therapeutics, against which several antibody clones have been developed and brought into therapeutic use. Another antibody clone, 528, is an antagonistic anti-EGFR antibody, which has been the focus of our antibody engineering studies to develop cancer drugs. In this study, we explored the interaction of 528 with the extracellular region of EGFR (sEGFR) via binding analyses and structural studies. Dot blotting experiments with heat treated sEGFR and surface plasmon resonance binding experiments revealed that 528 recognizes the tertiary structure of sEGFR and exhibits competitive binding to sEGFR with EGF and cetuximab. Single particle analysis of the sEGFR–528 Fab complex via electron microscopy clearly showed the binding of 528 to domain III of sEGFR, the domain to which EGF and cetuximab bind, explaining its antagonistic activity. Comparison between the two-dimensional class average and the cetuximab/sEGFR crystal structure revealed that 528 binds to a site that is shifted from, rather than identical to, the cetuximab epitope, and may exclude known drug-resistant EGFR mutations.
format	article
author	Koki Makabe Takeshi Yokoyama Shiro Uehara Tomomi Uchikubo-Kamo Mikako Shirouzu Kouki Kimura Kouhei Tsumoto Ryutaro Asano Yoshikazu Tanaka Izumi Kumagai
author_facet	Koki Makabe Takeshi Yokoyama Shiro Uehara Tomomi Uchikubo-Kamo Mikako Shirouzu Kouki Kimura Kouhei Tsumoto Ryutaro Asano Yoshikazu Tanaka Izumi Kumagai
author_sort	Koki Makabe
title	Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
title_short	Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
title_full	Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
title_fullStr	Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
title_full_unstemmed	Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope
title_sort	anti-egfr antibody 528 binds to domain iii of egfr at a site shifted from the cetuximab epitope
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/051aab10792249469899ed21bbfad1a2
work_keys_str_mv	AT kokimakabe antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT takeshiyokoyama antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT shirouehara antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT tomomiuchikubokamo antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT mikakoshirouzu antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT koukikimura antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT kouheitsumoto antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT ryutaroasano antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT yoshikazutanaka antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope AT izumikumagai antiegfrantibody528bindstodomainiiiofegfratasiteshiftedfromthecetuximabepitope
_version_	1718395800995758080

Anti-EGFR antibody 528 binds to domain III of EGFR at a site shifted from the cetuximab epitope

Ejemplares similares