title: An antibody with high reactivity for disease-associated α-synuclein reveals extensive brain pathology.
abstract: α-Synuclein is the major protein associated with Lewy body dementia, Parkinson's disease and multiple system atrophy. Since α-synuclein is present in the brain in physiological conditions as a presynaptic protein, it is crucial to characterize disease-associated modifications to develop an in vivo biomarker. With the aim to develop antibodies showing high specificity and sensitivity for disease-associated α-synuclein, synthetic peptides containing different amino acid sequences were used for immunization of mice. After generation of α-synuclein aggregates, ELISA and immunoblotting were used to test the specificity of antibodies. Tissue microarray sections originating from different human α-synucleinopathies were used to compare immunostaining with other, commercially available antibodies. Immunization of mice with the peptide TKEGVVHGVATVAE (amino acid 44-57 of α-synuclein) resulted in the generation of a monoclonal antibody (5G4), which was able to bind aggregated α-synuclein preparation in sandwich ELISA or coated on magnetic beads. 5G4 proved to be superior to other antibodies in comparative immunohistochemical studies by revealing more widespread and distinct α-synuclein pathology. Immunoblotting of human brain tissue revealed an additional band seen in dementia with Lewy bodies, whereas the band representing monomeric α-synuclein was very weak or lacking. In summary, the 5G4 antibody is most promising for re-evaluation of archival material and may offer new perspective for the development of in vivo diagnostic assays for detecting disease-associated α-synuclein in body fluids.
authors.affiliation: Institute of Neurology, Medical University of Vienna, AKH 4 J, Währinger Gürtel 18-20, 1097, Vienna, Austria. gabor.kovacs@meduniwien.ac.at
authors.forename: Gabor G
authors.initials: GG
authors.lastname: Kovacs
authors.forename: Uta
authors.initials: U
authors.lastname: Wagner
authors.forename: Benoit
authors.initials: B
authors.lastname: Dumont
authors.forename: Maria
authors.initials: M
authors.lastname: Pikkarainen
authors.forename: Awad A
authors.initials: AA
authors.lastname: Osman
authors.forename: Nathalie
authors.initials: N
authors.lastname: Streichenberger
authors.forename: Irene
authors.initials: I
authors.lastname: Leisser
authors.forename: Jérémy
authors.initials: J
authors.lastname: Verchère
authors.forename: Thierry
authors.initials: T
authors.lastname: Baron
authors.forename: Irina
authors.initials: I
authors.lastname: Alafuzoff
authors.forename: Herbert
authors.initials: H
authors.lastname: Budka
authors.forename: Armand
authors.initials: A
authors.lastname: Perret-Liaudet
authors.forename: Ingolf
authors.initials: I
authors.lastname: Lachmann
mesh_terms: D000906:Antibodies / Q000378:metabolism*; D001921:Brain / Q000378:metabolism* / Q000473:pathology*; D001927:Brain Diseases / Q000473:pathology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D057174:Frontotemporal Lobar Degeneration / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D019578:Multiple System Atrophy / Q000473:pathology; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D010300:Parkinson Disease / Q000473:pathology; D051844:alpha-Synuclein / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D051844:alpha-Synuclein