Generation and characterization of novel conformation-specific monoclonal antibodies for α-synuclein pathology

Neurobiol Dis. 2015 Jul;79:81-99. doi: 10.1016/j.nbd.2015.04.009. Epub 2015 Apr 30.

Abstract

α-Synuclein (α-syn), a small protein that has the intrinsic propensity to aggregate, is implicated in several neurodegenerative diseases including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), which are collectively known as synucleinopathies. Genetic, pathological, biochemical, and animal modeling studies provided compelling evidence that α-syn aggregation plays a key role in the pathogenesis of PD and related synucleinopathies. It is therefore of utmost importance to develop reliable tools that can detect the aggregated forms of α-syn. We describe here the generation and characterization of six novel conformation-specific monoclonal antibodies that recognize specifically α-syn aggregates but not the soluble, monomeric form of the protein. The antibodies described herein did not recognize monomers or fibrils generated from other amyloidogenic proteins including β-syn, γ-syn, β-amyloid, tau protein, islet amyloid polypeptide and ABri. Interestingly, the antibodies did not react to overlapping linear peptides spanning the entire sequence of α-syn, confirming further that they only detect α-syn aggregates. In immunohistochemical studies, the new conformation-specific monoclonal antibodies showed underappreciated small micro-aggregates and very thin neurites in PD and DLB cases that were not observed with generic pan antibodies that recognize linear epitope. Furthermore, employing one of our conformation-specific antibodies in a sandwich based ELISA, we observed an increase in levels of α-syn oligomers in brain lysates from DLB compared to Alzheimer's disease and control samples. Therefore, the conformation-specific antibodies portrayed herein represent useful tools for research, biomarkers development, diagnosis and even immunotherapy for PD and related pathologies.

Keywords: Conformation-specific monoclonal antibodies; Dementia with Lewy bodies; Parkinson’s disease; α-Synuclein.

Publication types

  • Multicenter Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Alzheimer Disease / metabolism
  • Amyloid beta-Peptides / metabolism
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Monoclonal / metabolism
  • Brain / metabolism*
  • Brain / pathology
  • Escherichia coli
  • Islet Amyloid Polypeptide / metabolism
  • Lewy Body Disease / metabolism
  • Lewy Body Disease / pathology
  • Membrane Glycoproteins / metabolism
  • Mice
  • Neoplasm Proteins / immunology
  • Neoplasm Proteins / metabolism
  • Parkinson Disease / metabolism
  • Parkinson Disease / pathology
  • Peptide Fragments / metabolism
  • Protein Conformation
  • Protein Multimerization
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Recombinant Proteins / metabolism
  • alpha-Synuclein / chemistry*
  • alpha-Synuclein / immunology*
  • alpha-Synuclein / metabolism
  • beta-Synuclein / immunology
  • beta-Synuclein / metabolism
  • gamma-Synuclein / immunology
  • gamma-Synuclein / metabolism
  • tau Proteins / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Amyloid beta-Peptides
  • Antibodies, Monoclonal
  • ITM2B protein, human
  • Islet Amyloid Polypeptide
  • MAPT protein, human
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • Peptide Fragments
  • Recombinant Proteins
  • SNCA protein, human
  • SNCB protein, human
  • SNCG protein, human
  • alpha-Synuclein
  • amyloid beta-protein (1-42)
  • beta-Synuclein
  • gamma-Synuclein
  • tau Proteins