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Open AccessArticle

Blood Contamination in CSF and Its Impact on Quantitative Analysis of Alpha-Synuclein

1
Ruhr-University, Faculty of Medicine, Medizinisches Proteom-Center, 44801 Bochum, Germany
2
University Medical Center Goettingen, Institute of Neuropathology, 37075 Goettingen, Germany
3
Department of Neurology, Ruhr-University Bochum at St Josef-Hospital, 44791 Bochum, Germany
4
Paracelsus-Elena Klinik, 34128 Kassel, Germany
5
University Medical Center Goettingen, Department of Neurology, 37075 Goettingen, Germany
*
Author to whom correspondence should be addressed.
Cells 2020, 9(2), 370; https://doi.org/10.3390/cells9020370
Received: 29 November 2019 / Revised: 24 January 2020 / Accepted: 28 January 2020 / Published: 5 February 2020
(This article belongs to the Special Issue The Molecular and Cellular Basis for Parkinson's Disease 2019)
Analysis of cerebrospinal fluid (CSF) is important for diagnosis of neurological diseases. Especially for neurodegenerative diseases, abnormal protein abundance in CSF is an important biomarker. However, the quality of CSF is a key factor for the analytic outcome. Any external contamination has tremendous impact on the analysis and the reliability of the results. In this study, we evaluated the effect of blood contamination in CSF with respect to protein biomarker identification. We compared three distinct measures: Combur10-Test® strips, a specific hemoglobin ELISA, and bottom-up mass spectrometry (MS)-based proteomics for the determination of the general blood contamination level. In parallel, we studied the impact of blood contamination on the detectability of alpha-synuclein (aSyn), a highly abundant protein in blood/erythrocytes and a potential biomarker for Parkinson’s disease. Comparable results were achieved, with all three approaches enabling detection of blood levels in CSF down to 0.001%. We found higher aSyn levels with increasing blood contamination, highlighting the difficulty of authentic quantification of this protein in CSF. Based on our results, we identified other markers for blood contamination beyond hemoglobin and defined a grading system for blood levels in CSF samples, including a lower limit of tolerable blood contamination for MS-based biomarker studies.
Keywords: cerebrospinal fluid; blood contamination; mass spectrometry; ELISA; urine strip; alpha-synuclein; quantification cerebrospinal fluid; blood contamination; mass spectrometry; ELISA; urine strip; alpha-synuclein; quantification
MDPI and ACS Style

Barkovits, K.; Kruse, N.; Linden, A.; Tönges, L.; Pfeiffer, K.; Mollenhauer, B.; Marcus, K. Blood Contamination in CSF and Its Impact on Quantitative Analysis of Alpha-Synuclein. Cells 2020, 9, 370.

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