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

MG Førland, A Öhrfelt, LS Oftedal… - Clinical Chemistry and …, 2017 - degruyter.com
MG Førland, A Öhrfelt, LS Oftedal, OB Tysnes, JP Larsen, K Blennow, H Zetterberg, G Alves
Clinical Chemistry and Laboratory Medicine (CCLM), 2017degruyter.com
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 …
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.
De Gruyter
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