Antibody-based methods for the measurement of α-synuclein concentration in human cerebrospinal fluid - method comparison and round robin study

J Neurochem. 2019 Apr;149(1):126-138. doi: 10.1111/jnc.14569. Epub 2018 Nov 13.

Abstract

α-Synuclein is the major component of Lewy bodies and a candidate biomarker for neurodegenerative diseases in which Lewy bodies are common, including Parkinson's disease and dementia with Lewy bodies. A large body of literature suggests that these disorders are characterized by reduced concentrations of α-synuclein in cerebrospinal fluid (CSF), with overlapping concentrations compared to healthy controls and variability across studies. Several reasons can account for this variability, including technical ones, such as inter-assay and inter-laboratory variation (reproducibility). We compared four immunochemical methods for the quantification of α-synuclein concentration in 50 unique CSF samples. All methods were designed to capture most of the existing α-synuclein forms in CSF ('total' α-synuclein). Each of the four methods showed high analytical precision, excellent correlation between laboratories (R2 0.83-0.99), and good correlation with each other (R2 0.64-0.93), although the slopes of the regression lines were different between the four immunoassays. The use of common reference CSF samples decreased the differences in α-synuclein concentration between detection methods and technologies. Pilot data on an immunoprecipitation mass spectrometry (IP-MS) method is also presented. Our results suggest that the four immunochemical methods and the IP-MS method measure similar forms of α-synuclein and that a common reference material would allow harmonization of results between immunoassays.

Keywords: biomarker; cerebrospinal fluid; enzyme-linked immunoabsorbent assay; mass spectrometry; round robin; α-synuclein.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / cerebrospinal fluid*
  • Female
  • Humans
  • Immunoassay / methods*
  • Lewy Body Disease / cerebrospinal fluid
  • Male
  • Multiple System Atrophy / cerebrospinal fluid
  • Parkinson Disease / cerebrospinal fluid
  • Reference Values
  • Reproducibility of Results
  • alpha-Synuclein / cerebrospinal fluid*

Substances

  • Biomarkers
  • alpha-Synuclein