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J Immunol Methods. 2015 Nov;426:70-5. doi: 10.1016/j.jim.2015.08.003. Epub 2015 Aug 10.

Validation of a commercially available enzyme-linked immunoabsorbent assay for the quantification of human α-Synuclein in cerebrospinal fluid.

Author information

1
Department of Neuropathology, University Medical Center, Robert-Koch-Str. 40, 37075 Göttingen, Germany. Electronic address: n.kruse@med.uni-goettingen.de.
2
Department of Neuropathology, University Medical Center, Robert-Koch-Str. 40, 37075 Göttingen, Germany; Paracelsus-Elena-Klinik, Klinikstr. 16, 34128 Kassel, Germany. Electronic address: brit.mollenhauer@paracelsus-kliniken.de.

Abstract

The quantification of α-Synuclein in cerebrospinal fluid (CSF) as a biomarker has gained tremendous interest in the last years. Several commercially available immunoassays are emerging. We here describe the full validation of one commercially available ELISA assay for the quantification of α-Synuclein in human CSF (Covance alpha-Synuclein ELISA kit). The study was conducted within the BIOMARKAPD project in the European initiative Joint Program for Neurodegenerative Diseases (JPND). We investigated the effect of several pre-analytical and analytical confounders: i.e. (1) need for centrifugation of freshly drawn CSF, (2) sample stability, (3) delay of freezing, (4) volume of storage aliquots, (5) freeze/thaw cycles, (6) thawing conditions, (7) dilution linearity, (8) parallelism, (9) spike recovery, and (10) precision. None of these confounders influenced the levels of α-Synuclein in CSF significantly. We found a very high intra-assay precision. The inter-assay precision was lower than expected due to different performances of kit lots used. Overall the validated immunoassay is useful for the quantification of α-Synuclein in human CSF.

KEYWORDS:

ELISA; Validation; alpha-Synuclein

PMID:
26271436
DOI:
10.1016/j.jim.2015.08.003
[Indexed for MEDLINE]

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