Enzyme-linked immunosorbent assays for α-synuclein with species and multimeric state specificities

J Neurosci Methods. 2011 Aug 15;199(2):249-57. doi: 10.1016/j.jneumeth.2011.05.020. Epub 2011 May 31.

Abstract

Abnormal intracellular deposition of aggregated α-synuclein is the characteristic feature of a number of neurological disorders, including Parkinson's disease (PD). Although α-synuclein is typically known as a cytosolic protein, a small amount is secreted by exocytosis in both monomeric and aggregated forms. The extracellular forms of α-synuclein in human body fluids, such as cerebrospinal fluid (CSF) and blood plasma, might be a diagnostic target for PD and related diseases. Here, we characterized a new set of monoclonal antibodies against α-synuclein, and using different combinations of antibodies, we established ELISA systems to specifically detect human α-synuclein, mouse and human α-synuclein together, and multimeric forms of α-synuclein in biological samples. By employing the Tyramide signal amplification method, the sensitivity of the assay was significantly improved to detect a concentration as low as ∼12.5 pg/ml. These assays might be useful tools for quantitative analysis of α-synuclein in various forms and with high sensitivity in diverse biological samples.

Publication types

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

MeSH terms

  • Adult
  • Animals
  • Antibodies, Monoclonal / isolation & purification
  • Antibody Specificity
  • COS Cells
  • Chlorocebus aethiops
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes / immunology*
  • Female
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Parkinson Disease / blood
  • Parkinson Disease / cerebrospinal fluid
  • Parkinson Disease / immunology
  • Protein Multimerization* / immunology
  • Young Adult
  • alpha-Synuclein / immunology
  • alpha-Synuclein / isolation & purification*
  • alpha-Synuclein / metabolism

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • alpha-Synuclein