Instability of cerebrospinal fluid after delayed storage and repeated freezing: a holistic study by drop coating deposition Raman spectroscopy

Jakub Klener; Kateřina Hofbauerová; Aleš Bartoš; Jan Říčný; Daniela Řípová; Vladimír Kopecký

doi:10.1515/cclm-2013-0800

Article

Instability of cerebrospinal fluid after delayed storage and repeated freezing: a holistic study by drop coating deposition Raman spectroscopy

Jakub Klener , Kateřina Hofbauerová , Aleš Bartoš , Jan Říčný , Daniela Řípová and Vladimír Kopecký

Published/Copyright: November 30, 2013

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From the journal Clinical Chemistry and Laboratory Medicine Volume 52 Issue 5

Abstract

Background: The handling of cerebrospinal fluid (CSF) affects the biomarker quantification used to diagnose Alzheimer’s disease (AD). Only specialized centers can test for AD markers. The precise timing and freezing is required to correctly measure these biomarkers. Therefore, the effects of CSF storage temperature and repeated freeze/thaw cycles on CSF stability were investigated.

Methods: Drop coating deposition Raman spectroscopy in combination with principal component analysis was used to analyze CSF and its dialyzed form (ELISA confirmed the removal of up to 80% of the AD markers). The advantage of this approach is that no prior knowledge of the biomarkers is necessary and that both the concentration and the protein structure of intact CSF are analyzed.

Results: Dialyzed CSF was stable for up to 5 h after its collection, while native CSF started to denature nearly immediately. Most of the unstable proteins were denatured within 24 h. The dialyzed CSF was not affected by freeze/thaw cycles, but the native CSF exhibited significant progressive changes, even after the first freezing. The mechanism as well as the resulting structures of the freeze-denatured proteins differed from those of the temporally denatured proteins, although both protein sets began with the same initial proteins.

Conclusions: CSF must be processed immediately, within 5 h of collection. Flash cooling is recommended for freezing CSF, but any freeze/thaw cycle will affect the protein component of CSF.

Keywords: Alzheimer’s disease; cerebrospinal fluid; cold denaturation; pre-analytical conditions; protein stability; Raman spectroscopy

Corresponding authors: Aleš Bartoš, Prague Psychiatric Center, Ústavní 91, 181 03 Prague 8, Bohnice, Czech Republic and Department of Neurology, Third Faculty of Medicine, University Hospital Královské Vinohrady, Charles University in Prague, Šrobárova 50, 100 34 Prague 10, Czech Republic, Phone: +420 266 003152, Fax: +420 266 003134, E-mail: bartos@pcp.lf3.cuni.cz; and Vladimír Kopecký Jr., Institute of Physics, Faculty of Mathematics and Physics, Charles University in Prague, Ke Karlovu 5, 121 16 Prague 2, Czech Republic, Phone: +420 221 911472, Fax: +420 224 922797, E-mail: kopecky@karlov.mff.cuni.cz

Acknowledgments

The authors appreciate the preliminary work of Nad’a Rosová (Charles University in Prague) on the DCDR spectroscopy of CSF during her bachelor’s studies. This work was supported by the Czech Ministry of Health (IGA, No. NT 13183), the Czech Science Foundation (No. 305/09/0457 and P304/12/G069) and the Academy of Sciences of the Czech Republic (RVO: 61388971).

Conflict of interest statement

Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research support played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

Research funding: None declared.

Employment or leadership: None declared.

Honorarium: None declared.

References

1. Hühmer AF, Bringer RG, Amato H, Fonteh AN, Harrington MG. Protein analysis in human cerebrospinal fluid: physiological aspects, current progress and future challenges. Dis Markers 2006;22:3–26.10.1155/2006/158797Search in Google Scholar PubMed PubMed Central

2. Deisenhammer F, Bartos A, Egg R, Gilhus NE, Giovannoni G, Rauer S, et al. Guidelines on routine cerebrospinal fluid analysis. Report from an EFNS task force. Eur J Neurol 2006;13:913–22.10.1111/j.1468-1331.2006.01493.xSearch in Google Scholar PubMed

3. Parada C, Parvas M, Bueno D. Cerebrospinal fluid proteomes: from neutral development to neurodegenerative diseases. Curr Proteomics 2007;4:89–106.10.2174/157016407782194611Search in Google Scholar

4. Schoonenboom NS, Mulder C, Vanderstichele H, Van Elk E-J, Kok A, Van Kamp GJ, et al. Effects of processing and storage conditions on amyloid β (1–42) and tau concentrations in cerebrospinal fluid: implications for use in clinical practice. Clin Chem 2005;51:189–95.10.1373/clinchem.2004.039735Search in Google Scholar PubMed

5. Kaiser E, Schönknecht P, Thomann PA, Hunt A, Schröder J. Influence of delayed CSF storage on concentrations of phospho-tau protein (181), total tau protein and beta-amyloid (1–42). Neurosci Lett 2007;417:193–5.10.1016/j.neulet.2007.02.045Search in Google Scholar PubMed

6. Zimmermann R, Lelental N, Ganslandt O, Maler JM, Kornhuber J, Lewczuk P. Preanalytical sample handling and sample stability testing for the neurochemical dementia diagnostics. J Alzheimers Dis 2011;25:739–45.10.3233/JAD-2011-110212Search in Google Scholar PubMed

7. Lewczuk P, Beck G, Ganslandt O, Esselmann H, Deisenhammer F, Regeniter A, et al. International quality control survey of neurochemical dementia diagnostics. Neurosci Lett 2006;409:1–4.10.1016/j.neulet.2006.07.009Search in Google Scholar PubMed

8. Varwey NA, van der Flier WM, Blennow K, Clark C, Sokolow S, De Deyn PP, et al. A worldwide multicentre comparison of assays for cerebrospinal fluid biomarkers in Alzheimer’s disease. Ann Clin Biochem 2009;46:235–40.10.1258/acb.2009.008232Search in Google Scholar PubMed

9. Mattsson N, Blennow K, Zetterber H. Inter-laboratory variation in cerebrospinal fluid biomarkers for Alzheimer’s disease: united we stand, divided we fall. Clin Chem Lab Med 2010;48:603–7.10.1515/CCLM.2010.131Search in Google Scholar PubMed

10. Rosenling T, Stoop MP, Smolinska A, Muilwijk B, Coulier L, Shi S, et al. The impact of delayed storage on the measured proteome and metabolome of human cerebrospinal fluid. Clin Chem 2011;57:1703–11.10.1373/clinchem.2011.167601Search in Google Scholar PubMed

11. Tensmeyer LG, Kauffman Jr EW. Protein structure as revealed by nonresonance Raman spectroscopy. In: Havel HA, editor. Spectroscopic methods for determining protein structure in solution. New York: VCH Publishers, 1996:69–95.Search in Google Scholar

12. Tuma R. Raman spectroscopy of proteins: from peptides to large assemblies. J Raman Spectrosc 2005;36:307–19.10.1002/jrs.1323Search in Google Scholar

13. Zhang D, Xie Y, Mrozek MF, Ortiz C, Davisson VJ, Ben-Amotz D. Raman detection of proteomic analytes. Anal Chem 2003;75:5703–9.10.1021/ac0345087Search in Google Scholar PubMed

14. Deegan RD, Bakajin O, Dupont TF, Huber G, Nagel SR, Witten TA. Capillary flow as the cause of ring stains from dried liquid drops. Nature 1997;389:827–9.10.1038/39827Search in Google Scholar

15. Kopecký Jr V, Baumruk V. Structure of the ring in drop coating deposited proteins and its implication for Raman spectroscopy of biomolecules. Vib Spectrosc 2006;42:184–7.10.1016/j.vibspec.2006.04.019Search in Google Scholar

16. Kapitán J, Baumruk V, Kopecký Jr V, Pohl R, Bouř P. Proline zwitterion dynamics in solution, glass, and crystalline state. J Am Chem Soc 2006;128:13451–62.10.1021/ja062958lSearch in Google Scholar PubMed

17. Ortiz C, Zhang D, Xie Y, Davisson VJ, Ben-Amotz D. Identification of insulin variants using Raman spectroscopy. Anal Biochem 2004;332:245–52.10.1016/j.ab.2004.06.013Search in Google Scholar PubMed

18. Xie Y, Zhang D, Jarori GK, Davisson VJ, Ben-Amotz D. The Raman detection of peptide tyroxine phosphorylation. Anal Biochem 2004;332:116–21.10.1016/j.ab.2004.05.052Search in Google Scholar PubMed

19. Filik J, Stone N. Analysis of human tear fluid by Raman spectroscopy. Anal Chim Acta 2008;616:177–84.10.1016/j.aca.2008.04.036Search in Google Scholar PubMed

20. Filik J, Stone N. Investigation into the protein composition of human tear fluid using centrifugal filters and drop coating deposition Raman spectroscopy. J Raman Spectrosc 2009;40:218–24.10.1002/jrs.2113Search in Google Scholar

21. Esmond-White KA, Mandair GS, Raaii F, Jacobson JA, Miller BS, Urquhart AG, et al. Raman spectroscopy of synovial fluid as a tool for diagnosing osteoarthritis. J Biomed Opt 2009;14:034013.10.1117/1.3130338Search in Google Scholar PubMed PubMed Central

22. Teunissen CE, Petzold A, Bennett JL, Berven FS, Brundin L, Comabella M, et al. A consensus protocol for the standardization of cerebrospinal fluid collection and biobanking. Neurology 2009;73:1914–22.10.1212/WNL.0b013e3181c47cc2Search in Google Scholar PubMed PubMed Central

23. Malinowski ER. Factor analysis in chemistry. Chichester: Wiley, 2002.Search in Google Scholar

24. Overman SA, Thomas Jr GJ. Raman markers of nonaromatic side chains in an α-helix assembly: Ala, Asp, Glu, Gly, Ile, Leu, Lys, Ser, and Val residuem of phage fd subunits. Biochemistry 1999;38:4018–27.10.1021/bi982901eSearch in Google Scholar PubMed

25. Slemmon JR, Meredith J, Guss V, Andreasson U, Andreasen N, Blennow K. Measurement of Aβ1–42 in cerebrospinal fluid is influenced by matrix effects. J Neurochem 2012;120:325–33.10.1111/j.1471-4159.2011.07553.xSearch in Google Scholar PubMed

26. Romero-Romero ML, Inglés-Prieto A, Ibarra-Molero B, Sanchez-Ruiz JM. Highly anamalous energetics of protein cold denaturation linked to folding-unfolding kinetics. PLoS One 2011;6:e23050.10.1371/journal.pone.0023050Search in Google Scholar PubMed PubMed Central

27. Alesci S, Borggrefe M, Dempfle CE. Effect of freezing method and storage at –20 °C and –70 °C on prothrombin time, aPPT and plasma fibrinogen levels. Thromb Res 2009;124: 121–6.10.1016/j.thromres.2008.11.010Search in Google Scholar PubMed

28. Griebe M, Daffertshofer M, Stroick M, Syren M, Ahmad-Nejad P, Neumaier M, et al. Infrared spectroscopy: a new diagnostic tool in Alzheimer disease. Neurosci Lett 2007;420:29–33.10.1016/j.neulet.2007.03.075Search in Google Scholar PubMed

Received: 2013-9-23

Accepted: 2013-10-20

Published Online: 2013-11-30

Published in Print: 2014-5-1

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https://doi.org/10.1515/cclm-2013-0800

Keywords for this article

Alzheimer’s disease; cerebrospinal fluid; cold denaturation; pre-analytical conditions; protein stability; Raman spectroscopy

Instability of cerebrospinal fluid after delayed storage and repeated freezing: a holistic study by drop coating deposition Raman spectroscopy

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Abstract

Acknowledgments

Conflict of interest statement

References

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