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Novel engineered nanobodies specific for N-terminal region of alpha-synuclein recognize Lewy-body pathology and inhibit in-vitro seeded aggregation and toxicity

Lookup NU author(s): Dr Daniel ErskineORCiD

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This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND).


Abstract

© 2022 The Authors. Nanobodies (Nbs), the single-domain antigen-binding fragments of dromedary heavy-chain antibodies (HCAb), are excellent candidates as therapeutic and diagnostic tools in synucleinopathies because of their small size, solubility and stability. Here, we constructed an immune nanobody library specific to the monomeric form of alpha-synuclein (α-syn). Phage display screening of the library allowed the identification of a nanobody, Nbα-syn01, specific for α-syn. Unlike previously developed nanobodies, Nbα-syn01 recognized the N-terminal region which is critical for in vitro and in vivo aggregation and contains many point mutations involved in early PD cases. The affinity of the monovalent Nbα-syn01 and the engineered bivalent format BivNbα-syn01 measured by isothermal titration calorimetry revealed unexpected results where Nbα-syn01 and its bivalent format recognized preferentially α-syn fibrils compared to the monomeric form. Nbα-syn01 and BivNbα-syn01 were also able to inhibit α-syn-seeded aggregation in vitro and reduced α-syn-seeded aggregation and toxicity in cells showing their potential to reduce α-syn pathology. Moreover, both nanobody formats were able to recognize Lewy-body pathology in human post-mortem brain tissue from PD and DLB cases. Additionally, we present evidence through structural docking that Nbα-syn01 binds the N-terminal region of the α-syn aggregated form. Overall, these results highlight the potential of Nbα-syn01 and BivNbα-syn01 in developing into a diagnostic or a therapeutic tool for PD and related disorders.


Publication metadata

Author(s): Hmila I, Vaikath NN, Majbour NK, Erskine D, Sudhakaran IP, Gupta V, Ghanem SS, Islam Z, Emara MM, Abdesselem HB, Kolatkar PR, Achappa DK, Vinardell T, El-Agnaf OMA

Publication type: Article

Publication status: Published

Journal: FEBS Journal

Year: 2022

Pages: Epub ahead of print

Online publication date: 28/01/2022

Acceptance date: 26/01/2022

Date deposited: 10/03/2022

ISSN (print): 1742-464X

ISSN (electronic): 1742-4658

Publisher: John Wiley and Sons Inc.

URL: https://doi.org/10.1111/febs.16376

DOI: 10.1111/febs.16376

PubMed id: 35090199


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Funding

Funder referenceFunder name
Qatar Biomedical Research Institute

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