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: Gabor G Kovacs, Uta Wagner, Benoit Dumont, Maria Pikkarainen, Awad A Osman, Nathalie Streichenberger, Irene Leisser, Jérémy Verchère, Thierry Baron, Irina Alafuzoff, Herbert Budka, Armand Perret-Liaudet, Ingolf Lachmann
• Journal: Acta neuropathologica (Acta Neuropathol) Vol. 124 Issue 1 Pg. 37-50 (Jul 2012) ISSN: 1432-0533 [Electronic] Germany
• PMID: 22370907 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies
• alpha-Synuclein

Topics

• Antibodies (metabolism)
• Brain (metabolism, pathology)
• Brain Diseases (pathology)
• Enzyme-Linked Immunosorbent Assay (methods)
• Frontotemporal Lobar Degeneration (pathology)
• Humans
• Immunohistochemistry
• Multiple System Atrophy (pathology)
• Neurons (metabolism, pathology)
• Parkinson Disease (pathology)
• alpha-Synuclein (immunology, metabolism)