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Acta Neuropathologica

, Volume 124, Issue 1, pp 37–50 | Cite as

An antibody with high reactivity for disease-associated α-synuclein reveals extensive brain pathology

  • Gabor G. KovacsEmail author
  • 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
Original Paper

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.

Keywords

α-Synuclein Biomarker Parkinson’s disease Lewy body dementia Multiple system atrophy 

Notes

Acknowledgments

This study was partly performed in the frame of the EU FP6 Project Neuroscreen LSHB-CZ-2006-037719 contract no. 037719. Patent pending for 5G4 antibody.

Supplementary material

401_2012_964_MOESM1_ESM.pdf (7.1 mb)
Supplementary material 1 (PDF 7237 kb)

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Gabor G. Kovacs
    • 1
    Email author
  • Uta Wagner
    • 2
  • Benoit Dumont
    • 3
    • 4
  • Maria Pikkarainen
    • 5
  • Awad A. Osman
    • 2
  • Nathalie Streichenberger
    • 6
  • Irene Leisser
    • 1
  • Jérémy Verchère
    • 7
  • Thierry Baron
    • 7
  • Irina Alafuzoff
    • 5
    • 8
  • Herbert Budka
    • 1
  • Armand Perret-Liaudet
    • 3
    • 4
  • Ingolf Lachmann
    • 2
  1. 1.Institute of Neurology, Medical University of ViennaViennaAustria
  2. 2.AJ Roboscreen GmbHLeipzigGermany
  3. 3.Neurobiology Laboratory, Department of BiochemistryCentre de Biologie et Pathologie Est, Hospices Civils de LyonLyonFrance
  4. 4.Centre de Recherche en Neurosciences de Lyon, Equipe BioRan Pr Luc ZimmerCNRS, INSERM, Claude Bernard Lyon1 UniversityLyonFrance
  5. 5.Institute of Clinical Medicine, NeurologyUniversity of Eastern FinlandKuopioFinland
  6. 6.Pathology and BiochemistryGroupement Hospitalier Est, Hospices Civils de Lyon/Claude Bernard UniversityLyonFrance
  7. 7.Agence Nationale de Sécurité Sanitaire (Anses)Unité Maladies Neuro-dégénérativesLyonFrance
  8. 8.Department of Immunology, Genetics and PathologyUppsala University and Uppsala University HospitalUppsalaSweden

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