A first tetraplex assay for the simultaneous quantification of total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in human cerebrospinal fluid

Niels Kruse, Michael G. Schlossmacher, Walter J. Schulz-Schaeffer, Eugeen Vanmechelen, Hugo Vanderstichele, Omar Ali El-Agnaf, Brit Mollenhauer

Research output: Contribution to journalArticle

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Abstract

The quantification of four distinct proteins (α-synuclein, β-amyloid1-42 , DJ-1, and total tau) in cerebrospinal fluid (CSF) has been proposed as a laboratory-based platform for the diagnosis of Parkinson's disease (PD) and Alzheimer's disease (AD). While there is some clinical utility in measuring these markers individually, their usage in routine clinical testing remains challenging, in part due to substantial overlap of concentrations between healthy controls and diseased subjects. In contrast, measurement of different analytes in a single sample from individual patients in parallel appears to considerably improve the accuracy of AD or PD diagnosis. Here, we report the development and initial characterization of a first, electrochemiluminescence-based multiplex immunoassay for the simultaneous quantification of all four proteins ('tetraplex') in as little as 50 μl of CSF. In analytical performance experiments, we assessed its sensitivity, spike-recovery rate, parallelism and dilution linearity as well as the intra- and inter-assay variability. Using our in-house calibrators, we recorded a lower limit of detection for α-synuclein, β-amyloid42 , DJ-1, and t-tau of 1.95, 1.24, 5.63, and 4.05 pg/ml, respectively. The corresponding, linear concentration range covered >3 orders of magnitude. In diluted CSF samples (up to 1:4), spike-recovery rates ranged from a low of 55% for β-amyloid42 to a high of 98% for DJ-1. Hillslopes ranged from 1.03 to 1.30, and inter-assay variability demonstrated very high reproducibility. Our newly established tetraplex assay represents a significant technical advance for fluid-based biomarker studies in neurodegenerative disorders allowing the simultaneous measurement of four pivotal makers in single CSF specimens. It provides exceptional sensitivity, accuracy and speed.

Original languageEnglish
Article numbere0153564
JournalPLoS One
Volume11
Issue number4
DOIs
Publication statusPublished - 1 Apr 2016

Fingerprint

Synucleins
Cerebrospinal fluid
cerebrospinal fluid
Cerebrospinal Fluid
Assays
Parkinson disease
assays
Alzheimer disease
Parkinson Disease
Alzheimer Disease
Recovery
neurodegenerative diseases
Biomarkers
disease diagnosis
immunoassays
Immunoassay
Neurodegenerative Diseases
reproducibility
Dilution
Limit of Detection

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kruse, N., Schlossmacher, M. G., Schulz-Schaeffer, W. J., Vanmechelen, E., Vanderstichele, H., Ali El-Agnaf, O., & Mollenhauer, B. (2016). A first tetraplex assay for the simultaneous quantification of total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in human cerebrospinal fluid. PLoS One, 11(4), [e0153564]. https://doi.org/10.1371/journal.pone.0153564

A first tetraplex assay for the simultaneous quantification of total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in human cerebrospinal fluid. / Kruse, Niels; Schlossmacher, Michael G.; Schulz-Schaeffer, Walter J.; Vanmechelen, Eugeen; Vanderstichele, Hugo; Ali El-Agnaf, Omar; Mollenhauer, Brit.

In: PLoS One, Vol. 11, No. 4, e0153564, 01.04.2016.

Research output: Contribution to journalArticle

Kruse, N, Schlossmacher, MG, Schulz-Schaeffer, WJ, Vanmechelen, E, Vanderstichele, H, Ali El-Agnaf, O & Mollenhauer, B 2016, 'A first tetraplex assay for the simultaneous quantification of total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in human cerebrospinal fluid', PLoS One, vol. 11, no. 4, e0153564. https://doi.org/10.1371/journal.pone.0153564
Kruse, Niels ; Schlossmacher, Michael G. ; Schulz-Schaeffer, Walter J. ; Vanmechelen, Eugeen ; Vanderstichele, Hugo ; Ali El-Agnaf, Omar ; Mollenhauer, Brit. / A first tetraplex assay for the simultaneous quantification of total α-Synuclein, Tau, β-Amyloid42 and DJ-1 in human cerebrospinal fluid. In: PLoS One. 2016 ; Vol. 11, No. 4.
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