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Pre-analytical factors influencing the stability of cerebrospinal fluid proteins

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Authors:
  • Simonsen, Anja H ;
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    Department of Clinical Medicine, Faculty of Health and Medical Sciences, Københavns Universitet
  • Bahl, Justyna M C ;
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    unknown
  • Danborg, Pia B ;
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    unknown
  • Lindstrom, Veronica ;
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    unknown
  • Larsen, Severin O ;
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    unknown
  • Grubb, Anders ;
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    unknown
  • Heegaard, Niels Henrik Helweg ;
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    unknown
  • Waldemar, Gunhild
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    Department of Clinical Medicine, Department of Clinical Medicine, Faculty of Health and Medical Sciences, Københavns Universitet
DOI:
10.1016/j.jneumeth.2013.03.011
Abstract:
Cerebrospinal fluid (CSF) is a potential source for new biomarkers due to its proximity to the brain. This study aimed to clarify the stability of the CSF proteome when undergoing pre-analytical factors. We investigated the effects of repeated freeze/thaw cycles, protease inhibitors and delayed storage for 4h, 24h or 14 days at -20°C, 4°C and room temperature (RT) after centrifugation compared with our standard practice of two hours at RT before placing the samples in an -80°C environment. The results were obtained using immunoassays for amyloid-beta 1-42 (Aβ42), tau, phosphorylated tau (P-tau) and cystatin C and using surface-enhanced laser desorption/ionisation time-of-flight (SELDI-TOF) mass spectrometry for proteomic profiling. Tau and P-tau were susceptible to repeated freeze/thaw cycles while SELDI-TOF analysis produced eight significant peaks and additional artefact peaks from samples with added protease inhibitors. Delayed storage for different durations and in different temperatures produced six significant SELDI-TOF peaks. Aβ42 and tau were susceptible to increased temperatures and the duration before storage, whereas P-tau and cystatin C were not. Transthyretin and several of its isoforms were found using SELDI-TOF and were susceptible to freeze/thaw cycles and to increased temperature and length of time prior to storage. We recommend that CSF should be collected and centrifuged immediately after sampling and prior to storage at -80°C without the addition of protease inhibitors. Freeze/thawing should be avoided because of the instability of tau, P-tau and transthyretin. Standardised CSF sampling, handling and storage for biomarker research are essential for accurately comparing the results obtained by different studies and institutions.
Type:
Journal article
Language:
English
Published in:
Journal of Neuroscience Methods, 2013, Vol 215, Issue 2, p. 234-240
Keywords:
Journal Article; Research Support, Non-U.S. Gov't
Main Research Area:
Medical science
Publication Status:
Published
Review type:
Peer Review
Submission year:
2013
Scientific Level:
Scientific
ID:
241288772

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