Abstract
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.
Highlights
A vast majority of studies done on neurological disorders use cerebrospinal fluid (CSF) as the biofluid of choice to explore and assess biomarkers as CSF protein levels most closely reflect the pathophysiology of the brain (Robey and Panegyres, 2019)
We aim to understand the stability of CSF α-syn biomarkers, mainly t-α-syn and o-α-syn, against potential preanalytical factors that occur during CSF handling and processing
Between lumbar puncture and laboratory analysis, collected CSF samples undergo variable sampling and storage methods, potentially different between individuals and sites processing the samples, which are often unknown to the scientists running the assays
Summary
A vast majority of studies done on neurological disorders use cerebrospinal fluid (CSF) as the biofluid of choice to explore and assess biomarkers as CSF protein levels most closely reflect the pathophysiology of the brain (Robey and Panegyres, 2019). Levels of different forms of α-syn in CSF have been widely investigated in several studies as potential biomarkers of synuclein aggregation disorders. The role of cerebrospinal fluid (CSF) alpha-synuclein as a potential biomarker has been challenged mainly due to variable preanalytical measures between laboratories. 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. 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
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