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The impact of delayed storage on the measured proteome and metabolome of human cerebrospinal fluid

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Author: Rosenling, T. · Stoop, M.P. · Smolinska, A. · Muilwijk, B. · Coulier, L. · Shi, S. · Dane, A. · Christin, C. · Suits, F. · Horvatovich, P.L. · Wijmenga, S.S. · Buydens, L.M.C. · Vreeken, R. · Hankemeier, T. · Gool, A.J. van · Luider, T.M. · Bischoff, R.
Source:Clinical Chemistry, 12, 57, 1703-1711
Identifier: 443695
doi: doi:10.1373/clinchem.2011.167601
Keywords: Nutrition · Healthy Living · Life Triskelion BV · QS - Quality & Safety AR - Analytical Research · EELS - Earth, Environmental and Life Sciences TNO Bedrijven


BACKGROUND: Because cerebrospinal fluid (CSF) is in close contact with diseased areas in neurological disorders, it is an important source of material in the search for molecular biomarkers. However, sample handling for CSF collected from patients in a clinical setting might not always be adequate for use in proteomics and metabolomics studies. METHODS: We left CSF for 0, 30, and 120 min at room temperature immediately after sample collection and centrifugation/removal of cells. At 2 laboratories CSF proteomes were subjected to tryptic digestion and analyzed by use of nano-liquid chromatography (LC) Orbitrap mass spectrometry (MS) and chipLC quadrupole TOF-MS. Metabolome analysis was performed at 3 laboratories by NMR, GC-MS, and LC-MS. Targeted analyses of cystatin C and albumin were performed by LC-tandem MS in the selected reaction monitoring mode. RESULTS: We did not find significant changes in the measured proteome and metabolome of CSF stored at room temperature after centrifugation, except for 2 peptides and 1 metabolite, 2,3,4-trihydroxybutanoic (threonic) acid, of 5780 identified peptides and 93 identified metabolites. A sensitive protein stability marker, cystatin C, was not affected. CONCLUSIONS: The measured proteome and metabolome of centrifuged human CSF is stable at room temperature for up to 2 hours. We cannot exclude, however, that changes undetectable with our current methodology, such as denaturation or proteolysis, might occur because of sample handling conditions. The stability we observed gives laboratory personnel at the collection site sufficient time to aliquot samples before freezing and storage at -80°C. © 2011 American Association for Clinical Chemistry.