Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor

Abstract Ligand-activated signaling through the type 1 insulin-like growth factor receptor (IGF1R) is implicated in many physiological processes ranging from normal human growth to cancer proliferation and metastasis. IGF1R has also emerged as a target for receptor-mediated transcytosis, a transport...

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Autores principales:	Joey Sheff, Ping Wang, Ping Xu, Melanie Arbour, Luke Masson, Henk van Faassen, Greg Hussack, Kristin Kemmerich, Eric Brunette, Danica Stanimirovic, Jennifer J. Hill, John Kelly, Feng Ni
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/abc8b420f9a841a1ad3480689cc4f109
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spelling	oai:doaj.org-article:abc8b420f9a841a1ad3480689cc4f1092021-12-02T12:11:28ZDefining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor10.1038/s41598-021-83198-w2045-2322https://doaj.org/article/abc8b420f9a841a1ad3480689cc4f1092021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-83198-whttps://doaj.org/toc/2045-2322Abstract Ligand-activated signaling through the type 1 insulin-like growth factor receptor (IGF1R) is implicated in many physiological processes ranging from normal human growth to cancer proliferation and metastasis. IGF1R has also emerged as a target for receptor-mediated transcytosis, a transport phenomenon that can be exploited to shuttle biotherapeutics across the blood–brain barrier (BBB). We employed differential hydrogen–deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) to characterize the interactions of the IGF1R ectodomain with a recently discovered BBB-crossing single-domain antibody (sdAb), VHH-IR5, in comparison with IGF-1 binding. HDX-MS confirmed that IGF-1 induced global conformational shifts in the L1/FnIII-1/-2 domains and α-CT helix of IGF1R. In contrast, the VHH-IR5 sdAb-mediated changes in conformational dynamics were limited to the α-CT helix and its immediate vicinity (L1 domain). High-resolution NMR spectroscopy titration data and linear peptide scanning demonstrated that VHH-IR5 has high-affinity binding interactions with a peptide sequence around the C-terminal region of the α-CT helix. Taken together, these results define a core linear epitope for VHH-IR5 within the α-CT helix, overlapping the IGF-1 binding site, and suggest a potential role for the α-CT helix in sdAb-mediated transcytosis.Joey SheffPing WangPing XuMelanie ArbourLuke MassonHenk van FaassenGreg HussackKristin KemmerichEric BrunetteDanica StanimirovicJennifer J. HillJohn KellyFeng NiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Joey Sheff Ping Wang Ping Xu Melanie Arbour Luke Masson Henk van Faassen Greg Hussack Kristin Kemmerich Eric Brunette Danica Stanimirovic Jennifer J. Hill John Kelly Feng Ni Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
description	Abstract Ligand-activated signaling through the type 1 insulin-like growth factor receptor (IGF1R) is implicated in many physiological processes ranging from normal human growth to cancer proliferation and metastasis. IGF1R has also emerged as a target for receptor-mediated transcytosis, a transport phenomenon that can be exploited to shuttle biotherapeutics across the blood–brain barrier (BBB). We employed differential hydrogen–deuterium exchange mass spectrometry (HDX-MS) and nuclear magnetic resonance (NMR) to characterize the interactions of the IGF1R ectodomain with a recently discovered BBB-crossing single-domain antibody (sdAb), VHH-IR5, in comparison with IGF-1 binding. HDX-MS confirmed that IGF-1 induced global conformational shifts in the L1/FnIII-1/-2 domains and α-CT helix of IGF1R. In contrast, the VHH-IR5 sdAb-mediated changes in conformational dynamics were limited to the α-CT helix and its immediate vicinity (L1 domain). High-resolution NMR spectroscopy titration data and linear peptide scanning demonstrated that VHH-IR5 has high-affinity binding interactions with a peptide sequence around the C-terminal region of the α-CT helix. Taken together, these results define a core linear epitope for VHH-IR5 within the α-CT helix, overlapping the IGF-1 binding site, and suggest a potential role for the α-CT helix in sdAb-mediated transcytosis.
format	article
author	Joey Sheff Ping Wang Ping Xu Melanie Arbour Luke Masson Henk van Faassen Greg Hussack Kristin Kemmerich Eric Brunette Danica Stanimirovic Jennifer J. Hill John Kelly Feng Ni
author_facet	Joey Sheff Ping Wang Ping Xu Melanie Arbour Luke Masson Henk van Faassen Greg Hussack Kristin Kemmerich Eric Brunette Danica Stanimirovic Jennifer J. Hill John Kelly Feng Ni
author_sort	Joey Sheff
title	Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
title_short	Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
title_full	Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
title_fullStr	Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
title_full_unstemmed	Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
title_sort	defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/abc8b420f9a841a1ad3480689cc4f109
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Defining the epitope of a blood–brain barrier crossing single domain antibody specific for the type 1 insulin-like growth factor receptor

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