| DOI | Trouver le DOI : https://doi.org/10.1093/jb/mvac088 |
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| Auteur | Rechercher : Sheff, Joey1Identifiant ORCID : https://orcid.org/0000-0002-9269-6863; Rechercher : Kelly, John1; Rechercher : Foss, Mary1; Rechercher : Brunette, Eric1; Rechercher : Kemmerich, Kristin1; Rechercher : van Faassen, Henk1; Rechercher : Raphael, Shalini1; Rechercher : Hussack, Greg1; Rechercher : Comamala, Gerard; Rechercher : Rand, Kasper; Rechercher : Stanimirovic, Danica B.1 |
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| Affiliation | - Conseil national de recherches du Canada. Thérapeutique en santé humaine
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| Format | Texte, Article |
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| Sujet | blood–brain barrier; epitope mapping; hydrogen–deuterium exchange mass spectrometry; IGF1R; single-domain antibody |
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| Résumé | Pathologies of the central nervous system impact a significant portion of our population, and the delivery of therapeutics for effective treatment is challenging. The insulin-like growth factor-1 receptor (IGF1R) has emerged as a target for receptor-mediated transcytosis, a process by which antibodies are shuttled across the blood–brain barrier (BBB). Here, we describe the biophysical characterization of VHH-IR4, a BBB-crossing single-domain antibody (sdAb). Binding was confirmed by isothermal titration calorimetry and an epitope was highlighted by surface plasmon resonance that does not overlap with the IGF-1 binding site or other known BBB-crossing sdAbs. The epitope was mapped with a combination of linear peptide scanning and hydrogen–deuterium exchange mass spectrometry (HDX-MS). IGF1R is large and heavily disulphide bonded, and comprehensive HDX analysis was achieved only through the use of online electrochemical reduction coupled with a multiprotease approach, which identified an epitope for VHH-IR4 within the cysteine-rich region (CRR) of IGF1R spanning residues W244-G265. This is the first report of an sdAb binding the CRR. We show that VHH-IR4 inhibits ligand induced auto-phosphorylation of IGF1R and that this effect is mediated by downstream conformational effects. Our results will guide the selection of antibodies with improved trafficking and optimized IGF1R binding characteristics. |
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| Date de publication | 2022-11-08 |
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| Maison d’édition | Oxford Academic |
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| Dans | |
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| Langue | anglais |
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| Publications évaluées par des pairs | Oui |
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| Exporter la notice | Exporter en format RIS |
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| Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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| Identificateur de l’enregistrement | 69c3e110-3e98-49f5-96f6-669c4cfca754 |
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| Enregistrement créé | 2023-09-14 |
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| Enregistrement modifié | 2023-09-14 |
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