Advanced

Pre-analytical factors influencing the stability of cerebrospinal fluid proteins

Simonsen, Anja H. ; Bahl, Justyna M. C. ; Danborg, Pia B. ; Lindström, Veronica LU ; Larsen, Severin O. ; Grubb, Anders LU ; Heegaard, Niels H. H. and Waldemar, Gunhild (2013) In Journal of Neuroscience Methods 215(2). p.234-240
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 4 h, 24 h or 14 days at -20 degrees C, 4 degrees C and room temperature (RT) after centrifugation compared with our standard practice of two hours at RT before placing the samples in an -80 degrees C environment. The results were obtained using immunoassays for amyloid-beta 1-42 (A beta 42), tau, phosphorylated tau (P-tau) and cystatin C and using surface-enhanced laser desorption/ionisation time-of-flight (SELDI-TOF) mass spectrometry... (More)
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 4 h, 24 h or 14 days at -20 degrees C, 4 degrees C and room temperature (RT) after centrifugation compared with our standard practice of two hours at RT before placing the samples in an -80 degrees C environment. The results were obtained using immunoassays for amyloid-beta 1-42 (A beta 42), 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. A beta 42 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 degrees 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. (c) 2013 Elsevier B.V. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cerebrospinal fluid, Proteome, Pre-analytical factors, SELDI-TOF, Immunoassays, Stability
in
Journal of Neuroscience Methods
volume
215
issue
2
pages
234 - 240
publisher
Elsevier
external identifiers
  • wos:000319957800009
  • scopus:84877148563
  • pmid:23537933
ISSN
1872-678X
DOI
10.1016/j.jneumeth.2013.03.011
language
English
LU publication?
yes
id
4c0c3c81-804a-49a9-86be-6e0d45887e78 (old id 3927358)
date added to LUP
2016-04-01 10:34:31
date last changed
2019-10-23 01:32:09
@article{4c0c3c81-804a-49a9-86be-6e0d45887e78,
  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 4 h, 24 h or 14 days at -20 degrees C, 4 degrees C and room temperature (RT) after centrifugation compared with our standard practice of two hours at RT before placing the samples in an -80 degrees C environment. The results were obtained using immunoassays for amyloid-beta 1-42 (A beta 42), 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. A beta 42 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 degrees 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. (c) 2013 Elsevier B.V. All rights reserved.},
  author       = {Simonsen, Anja H. and Bahl, Justyna M. C. and Danborg, Pia B. and Lindström, Veronica and Larsen, Severin O. and Grubb, Anders and Heegaard, Niels H. H. and Waldemar, Gunhild},
  issn         = {1872-678X},
  language     = {eng},
  number       = {2},
  pages        = {234--240},
  publisher    = {Elsevier},
  series       = {Journal of Neuroscience Methods},
  title        = {Pre-analytical factors influencing the stability of cerebrospinal fluid proteins},
  url          = {http://dx.doi.org/10.1016/j.jneumeth.2013.03.011},
  doi          = {10.1016/j.jneumeth.2013.03.011},
  volume       = {215},
  year         = {2013},
}