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

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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.

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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.

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Authors and Affiliations

  1. Institute of Neurology, Medical University of Vienna, AKH 4 J, Währinger Gürtel 18-20, 1097, Vienna, Austria

    Gabor G. Kovacs, Irene Leisser & Herbert Budka

  2. AJ Roboscreen GmbH, Delitzscher Strasse 135, 04129, Leipzig, Germany

    Uta Wagner, Awad A. Osman & Ingolf Lachmann

  3. Neurobiology Laboratory, Department of Biochemistry, Centre de Biologie et Pathologie Est, Hospices Civils de Lyon, Lyon, France

    Benoit Dumont & Armand Perret-Liaudet

  4. Centre de Recherche en Neurosciences de Lyon, Equipe BioRan Pr Luc Zimmer, CNRS, INSERM, Claude Bernard Lyon1 University, Lyon, France

    Benoit Dumont & Armand Perret-Liaudet

  5. Institute of Clinical Medicine, Neurology, University of Eastern Finland, Kuopio, Finland

    Maria Pikkarainen & Irina Alafuzoff

  6. Pathology and Biochemistry, Groupement Hospitalier Est, Hospices Civils de Lyon/Claude Bernard University, Lyon, France

    Nathalie Streichenberger

  7. Agence Nationale de Sécurité Sanitaire (Anses), Unité Maladies Neuro-dégénératives, Lyon, France

    Jérémy Verchère & Thierry Baron

  8. Department of Immunology, Genetics and Pathology, Uppsala University and Uppsala University Hospital, Uppsala, Sweden

    Irina Alafuzoff

Authors
  1. Gabor G. Kovacs
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  2. Uta Wagner
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  3. Benoit Dumont
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  7. Irene Leisser
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  8. Jérémy Verchère
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  9. Thierry Baron
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  10. Irina Alafuzoff
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  11. Herbert Budka
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  12. Armand Perret-Liaudet
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  13. Ingolf Lachmann
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Corresponding author

Correspondence to Gabor G. Kovacs.

Additional information

The authors G. G. Kovacs and I. Lachmann contributed equally.

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Kovacs, G.G., Wagner, U., Dumont, B. et al. An antibody with high reactivity for disease-associated α-synuclein reveals extensive brain pathology. Acta Neuropathol 124, 37–50 (2012). https://doi.org/10.1007/s00401-012-0964-x

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  • DOI: https://doi.org/10.1007/s00401-012-0964-x

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