title: Antibodies against the C-terminus of α-synuclein modulate its fibrillation.
abstract: The 140-residue natively disordered protein α-synuclein (aSN) is a central component in the development of a family of neurodegenerative diseases termed synucleinopathies. This is attributed to its ability to form cytotoxic aggregates such as oligomers and amyloid fibrils. Consequently there have been intense efforts to avoid aggregation or reroute the aggregation pathway using pharmaceutical agents such as small molecules, chaperones and antibodies. aSN's lack of persistent structure in the monomeric state, as well as the multitude of different oligomeric and even different fibrillar states, makes it difficult to raise antibodies that would be efficacious in neutralizing all conformations of aSN. However, the C-terminal 20-40 residues of aSN are a promising epitope for antibody development. It is primarily disordered in both monomeric and aggregated forms, and an anti-C-terminal antibody will therefore be able to bind all forms. Furthermore, it might not interfere with the folding of aSN into membranes, which could be important for its physiological role. Here we report a screen of a series of monoclonal antibodies, which all target the C-terminal of aSN. According to dot blot analyses, different antibodies bound different forms of aSN with different preferences and showed reduced binding to monomeric compared to aggregated (oligomeric and fibrillary) aSN. Consequently they have different effects on aSN's ability to fibrillate and permeabilize membranes. Generally, the antibodies with strongest binding to aggregated aSN in dot blot, also inhibited fibrillation and membrane permeabilization the most, and promoted formation of amorphous aggregates surrounded by small and thin fibers. This suggests that the development of antibodies that targets the C-terminus, exposed in the aggregated forms of aSN, may be beneficial for improved immunotherapy against PD.
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
authors.forename: Cagla
authors.initials: C
authors.lastname: Sahin
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Large Protein Biophysics and Formulation, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark.
authors.forename: Nikolai
authors.initials: N
authors.lastname: Lorenzen
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark.
authors.forename: Lasse
authors.initials: L
authors.lastname: Lemminger
authors.affiliation: Department of Biomedicine, Aarhus University, Wilhelm Meyers Allé 4, DK-8000 Aarhus, Denmark.
authors.forename: Gunna
authors.initials: G
authors.lastname: Christiansen
authors.affiliation: Department of Molecular Biology and Genetics, Forsøgsvej 1, DK-4200 Slagelse, Denmark.
authors.forename: Ian Max
authors.initials: IM
authors.lastname: Møller
authors.affiliation: Division of Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej, DK-2500 Valby, Denmark.
authors.forename: Louise Buur
authors.initials: LB
authors.lastname: Vesterager
authors.affiliation: Division of Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej, DK-2500 Valby, Denmark.
authors.forename: Lars Østergaard
authors.initials: LØ
authors.lastname: Pedersen
authors.affiliation: Division of Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej, DK-2500 Valby, Denmark.
authors.forename: Karina
authors.initials: K
authors.lastname: Fog
authors.affiliation: Division of Neurodegeneration and Biologics, H. Lundbeck A/S, Ottiliavej, DK-2500 Valby, Denmark.
authors.forename: Pekka
authors.initials: P
authors.lastname: Kallunki
authors.affiliation: Interdisciplinary Nanoscience Center (iNANO), Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark; Department of Molecular Biology and Genetics, Aarhus University, Gustav Wieds Vej 14, DK-8000 Aarhus C, Denmark. Electronic address: dao@inano.au.dk.
authors.forename: Daniel E
authors.initials: DE
authors.lastname: Otzen
mesh_terms: D000682:Amyloid / Q000187:drug effects*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D006801:Humans; D051379:Mice; D010300:Parkinson Disease / Q000188:drug therapy; D066329:Protein Aggregates / Q000187:drug effects; D066263:Protein Aggregation, Pathological / Q000188:drug therapy / Q000517:prevention & control*; D051844:alpha-Synuclein / Q000276:immunology*
chemical_list: D000682:Amyloid; D000911:Antibodies, Monoclonal; D066329:Protein Aggregates; D051844:alpha-Synuclein
keywords: Amyloid; Cytotoxic oligomers; Fibrillation inhibition; Immunotherapy; Parkinson's disease