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Validation of a new assay for alpha-synuclein detection in cerebrospinal fluid

Forland, MG; Ohrfelt, A; Oftedal, LS; Tysnes, O-B; Larsen, JP; Blennow, K; Zetterberg, H; ... Lange, J; + view all (2017) Validation of a new assay for alpha-synuclein detection in cerebrospinal fluid. Clinical Chemistry and Laboratory Medicine , 55 (2) pp. 254-260. 10.1515/cclm-2016-0409. Green open access

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Abstract

BACKGROUND: Abnormal α-synuclein aggregation and deposition is the pathological hallmark of Parkinson’s disease (PD) and dementia with Lewy bodies (DLB), but is also found in Alzheimer disease (AD). Therefore, there is a gaining interest in α-synuclein in cerebrospinal fluid (CSF) as potential biomarker for these neurodegenerative diseases. To broaden the available choices of α-synuclein measurement in CSF, we developed and validated a new assay for detecting total α-synuclein. METHODS: This novel ELISA uses commercially available antibodies and is based on electrochemiluminescence technology. The assay protocol is straightforward, with short and simple incubation steps, and requires only small amounts of CSF. We validated this assay for precision, parallelism, dilution linearity, specificity, and spike recovery. We further compared it to the newly validated α-synuclein assay from BioLegend by analyzing a set of 50 CSF samples with both assays. RESULTS: The new assay quantifies α-synuclein in CSF with a lower limit of detection of 36.3 pg/mL and shows no cross-reactivity with human β- and γ-synuclein. Results of dilution linearity, parallelism, spike recovery, and precision classify this assay as well suited for α-synuclein detection in human CSF samples. CONCLUSIONS: We present a novel assay based on freely available components to quantify total α-synuclein in CSF as an additional method for α-synuclein as a biomarker in neurodegenerative diseases. The assay convinces with its simple and convenient protocol paired with high sensitivity.

Type: Article
Title: Validation of a new assay for alpha-synuclein detection in cerebrospinal fluid
Open access status: An open access version is available from UCL Discovery
DOI: 10.1515/cclm-2016-0409
Publisher version: https://doi.org/10.1515/cclm-2016-0409
Language: English
Additional information: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
Keywords: Alpha-synuclein, assay validation, biomarker, cerebrospinal fluid, elisa, alzheimers-disease, lewy bodies, neurodegenerative disorders, parkinsons-disease, csf, biomarkers, dementia, quantification
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Neurodegenerative Diseases
URI: https://discovery.ucl.ac.uk/id/eprint/1537981
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