Preanalytical Stability of CSF Total and Oligomeric Alpha-Synuclein

Background: The role of cerebrospinal fluid (CSF) alpha-synuclein as a potential biomarker has been challenged mainly due to variable preanalytical measures between laboratories. To evaluate the impact of the preanalytical factors contributing to such variability, the different subforms of alpha-synuclein need to be studied individually. Method: We investigated the effect of exposing CSF samples to several preanalytical sources of variability: (1) different polypropylene (PP) storage tubes; (2) use of non-ionic detergents; (3) multiple tube transfers; (4) multiple freeze-thaw cycles; and (5) delayed storage. CSF oligomeric- and total-alpha-synuclein levels were estimated using our in-house sandwich-based enzyme-linked immunosorbent assays. Results: Siliconized tubes provided the optimal preservation of CSF alpha-synuclein proteins among other tested polypropylene tubes. The use of tween-20 detergent significantly improved the recovery of oligomeric-alpha-synuclein, while multiple freeze-thaw cycles significantly lowered oligomeric-alpha-synuclein in CSF. Interestingly, oligomeric-alpha-synuclein levels remained relatively stable over multiple tube transfers and upon delayed storage. Conclusion: Our study showed for the first-time distinct impact of preanalytical factors on the different forms of CSF alpha-synuclein. These findings highlight the need for special considerations for the different forms of alpha-synuclein during CSF samples’ collection and processing.

[1]  W. Poewe,et al.  Safety and immunogenicity of the α-synuclein active immunotherapeutic PD01A in patients with Parkinson's disease: a randomised, single-blinded, phase 1 trial , 2020, The Lancet Neurology.

[2]  H. Berendse,et al.  CSF total and oligomeric α-Synuclein along with TNF-α as risk biomarkers for Parkinson’s disease: a study in LRRK2 mutation carriers , 2020, Translational Neurodegeneration.

[3]  B. Mollenhauer,et al.  Blood Contamination in CSF and Its Impact on Quantitative Analysis of Alpha-Synuclein , 2020, Cells.

[4]  W. M. van der Flier,et al.  Pre-analytical stability of novel cerebrospinal fluid biomarkers. , 2019, Clinica chimica acta; international journal of clinical chemistry.

[5]  Paolo Eusebi,et al.  CSF and blood biomarkers for Parkinson's disease , 2019, The Lancet Neurology.

[6]  J. Trojanowski,et al.  Impact of Pre-Analytical Differences on Biomarkers in the ADNI and PPMI Studies: Implications in the Era of Classifying Disease Based on Biomarkers. , 2019, Journal of Alzheimer's disease : JAD.

[7]  S. Lehmann,et al.  Impact of CSF storage volume on the analysis of Alzheimer's disease biomarkers on an automated platform. , 2019, Clinica chimica acta; international journal of clinical chemistry.

[8]  P. Panegyres,et al.  Cerebrospinal fluid biomarkers in neurodegenerative disorders , 2019, Future Neurology.

[9]  K. Blennow,et al.  Antibody‐based methods for the measurement of α‐synuclein concentration in human cerebrospinal fluid – method comparison and round robin study , 2018, Journal of neurochemistry.

[10]  D. Wales,et al.  Mapping Surface Hydrophobicity of α-Synuclein Oligomers at the Nanoscale , 2018, Nano letters.

[11]  Leslie M. Shaw,et al.  Longitudinal CSF biomarkers in patients with early Parkinson disease and healthy controls , 2017, Neurology.

[12]  P. Calabresi,et al.  Diagnostic utility of cerebrospinal fluid α‐synuclein in Parkinson's disease: A systematic review and meta‐analysis , 2017, Movement disorders : official journal of the Movement Disorder Society.

[13]  D. Selkoe,et al.  A user's guide for α‐synuclein biomarker studies in biological fluids: Perianalytical considerations , 2017, Movement disorders : official journal of the Movement Disorder Society.

[14]  H. Vanderstichele,et al.  Optimized Standard Operating Procedures for the Analysis of Cerebrospinal Fluid Aβ42 and the Ratios of Aβ Isoforms Using Low Protein Binding Tubes , 2016, Journal of Alzheimer's disease : JAD.

[15]  Arthur W. Toga,et al.  CSF biomarkers associated with disease heterogeneity in early Parkinson’s disease: the Parkinson’s Progression Markers Initiative study , 2016, Acta Neuropathologica.

[16]  Xue Ming,et al.  Respiratory and autonomic dysfunction in children with autism spectrum disorders , 2016, Brain and Development.

[17]  Takahiko Tokuda,et al.  Oligomeric and phosphorylated alpha-synuclein as potential CSF biomarkers for Parkinson’s disease , 2016, Molecular Neurodegeneration.

[18]  Stephanie M. Williams,et al.  Oligomeric α‐synuclein and β‐amyloid variants as potential biomarkers for Parkinson's and Alzheimer's diseases , 2016, The European journal of neuroscience.

[19]  P. Calabresi,et al.  Differential role of CSF alpha-synuclein species, tau, and Aβ42 in Parkinson's Disease , 2014, Front. Aging Neurosci..

[20]  W. Guder.  History of the preanalytical phase: a personal view , 2014, Biochemia medica.

[21]  P. Calabresi,et al.  Cerebrospinal Fluid Lysosomal Enzymes and Alpha-Synuclein in Parkinson's Disease , 2014, Movement disorders : official journal of the Movement Disorder Society.

[22]  B. Mollenhauer,et al.  Total CSF α-synuclein is lower in de novo Parkinson patients than in healthy subjects , 2013, Neuroscience Letters.

[23]  J. Molinuevo,et al.  Recommendations to standardize preanalytical confounding factors in Alzheimer's and Parkinson's disease cerebrospinal fluid biomarkers: an update. , 2012, Biomarkers in medicine.

[24]  Vladimir N Uversky,et al.  Α-synuclein misfolding and Parkinson's disease. , 2012, Biochimica et biophysica acta.

[25]  S. Cheon,et al.  Elevated Levels of α-Synuclein Oligomer in the Cerebrospinal Fluid of Drug-Naïve Patients with Parkinson's Disease , 2011, Journal of clinical neurology.

[26]  P. Calabresi,et al.  Cerebrospinal fluid Tau/α‐synuclein ratio in Parkinson's disease and degenerative dementias , 2011, Movement disorders : official journal of the Movement Disorder Society.

[27]  O. Laterza,et al.  Nonspecific binding of Aβ42 to polypropylene tubes and the effect of Tween-20. , 2010, Clinica chimica acta; international journal of clinical chemistry.

[28]  B. Mollenhauer,et al.  Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration , 2008, Experimental Neurology.

[29]  Juan Manuel Maler,et al.  Effect of sample collection tubes on cerebrospinal fluid concentrations of tau proteins and amyloid beta peptides. , 2006, Clinical chemistry.

[30]  M. Brook,et al.  Silicone elastomers for reduced protein adsorption. , 2004, Biomaterials.

[31]  D. Walsh,et al.  Soluble oligomers for the diagnosis of neurodegenerative diseases , 2003, The Lancet Neurology.

[32]  Nigel J. Cairns,et al.  Filamentous α-synuclein inclusions link multiple system atrophy with Parkinson's disease and dementia with Lewy bodies , 1998, Neuroscience Letters.

[33]  H. Vanderstichele,et al.  Risk of Alzheimer's disease biological misdiagnosis linked to cerebrospinal collection tubes. , 2012, Journal of Alzheimer's disease : JAD.