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Accuracy of cerebrospinal fluid Aβ1-42 measurements: evaluation of pre-analytical factors using a novel Elecsys immunosassay

  • Malgorzata Rozga ORCID logo EMAIL logo , Tobias Bittner , Kina Höglund and Kaj Blennow
Published/Copyright: February 4, 2017
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 55 Issue 10

Abstract

Background:

A decreased level of Aβ1-42 in cerebrospinal fluid (CSF) is characteristic of Alzheimer disease and often used to support clinical diagnosis. The measured concentration of CSF Aβ1-42, however, depends strongly on several pre-analytical and analytical “confounding” factors such as sample collection, material of testing tube, CSF handling and storage procedures (e.g. transfer to new tubes after centrifugation, freeze-thaw effects). As a consequence, substantial variations in the measured levels of this biomarker are observed even for the same sample. This study investigates whether the accuracy of quantitative analysis of CSF Aβ1-42 can be improved by pre-analytical treatment of CSF with agents that could potentially reduce a freeze-thaw and adhesion-related depletion of Aβ1-42 from CSF, including modulators of Aβ aggregation and cryoprotecting or anti-adhesion agents.

Methods:

The concentration of CSF Aβ1-42 was assessed with a novel Elecsys immunoassay developed for quantification of Aβ1-42 in human CSF.

Results:

Low-molecular weight Aβ oligomerization inhibitors, β-sheet breaker peptides, or the mid domain 4G8 antibody do not improve the stability of CSF Aβ1-42 during a repeated freeze-thaw treatment. Cryoprotecting agents reduce a freeze-thaw dependent loss of Aβ1-42 only when spiked to CSF to final concentration of 300 mM or higher. Adhesion of Aβ1-42 can be prevented by pre-treating CSF with Tween or by using tubes with a siliconized surface.

Conclusions:

Between-center variability in measured level of CSF Aβ1-42 can be reduced only by standardized CSF collection into one specific tube that, without centrifugation, transfer or other types of pre-analytical processing, is directly analyzed after sample collection.

Keywords: Aβ-peptide; Alzheimer disease; cerebrospinal fluid; Elecsys β-amyloid (1-42) assay
Corresponding author: Malgorzata Rozga, PhD, Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at University of Gothenburg, House V3/SU, SE-431 80 Mölndal, Sweden, Phone: +46 31 343 24 41, Fax: +46 31 41 92 89.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) 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.

References

1. Blennow K, Hampel H, Weiner M, Zetterberg H. Cerebrospinal fluid and plasma biomarkers in Alzheimer disease. Nat Rev Neurol 2010;6:131–44.10.1038/nrneurol.2010.4Search in Google Scholar

2. Blennow K, Mattsson N, Scholl M, Hansson O, Zetterberg H. Amyloid biomarkers in Alzheimer's disease. Trends Pharmacol Sci 2015;36:297–309.10.1016/j.tips.2015.03.002Search in Google Scholar

3. Blennow K, Vanmechelen E, Hampel H. CSF total tau, a beta 42 and phosphorylated tau protein as biomarkers for Alzheimer's disease. Mol Neurobiol 2001;24:87–97.10.1385/MN:24:1-3:087Search in Google Scholar

4. Ahmed M, Davis J, Aucoin D, Sato T, Ahuja S, Aimoto S, et al. Structural conversion of neurotoxic amyloid-beta(1-42) oligomers to fibrils. Nat Struct Mol Biol 2010;17:561–7.10.1038/nsmb.1799Search in Google Scholar

5. Rozga M, Kloniecki M, Jablonowska A, Dadlez M, Bal W. The binding constant for amyloid Abeta40 peptide interaction with human serum albumin. Biochem Biophys Res Commun 2007;364:714–8.10.1016/j.bbrc.2007.10.080Search in Google Scholar

6. Du YS, Ni BH, Glinn M, Dodel RC, Bales KR, Zhang ZY, et al. alpha(2)-macroglobulin as a beta-amyloid peptide-binding plasma protein. J Neurochem 1997;69:299–305.10.1046/j.1471-4159.1997.69010299.xSearch in Google Scholar

7. Matsubara E, Frangione B, Ghiso J. Characterization of apolipoprotein J-Alzheimer’s A beta interaction. J Biol Chem 1995;270:7563–7.10.1074/jbc.270.13.7563Search in Google Scholar

8. Schwarzman AL, Gregori L, Vitek MP, Lyubski S, Strittmatter WJ, Enghilde JJ, et al. Transthyretin sequesters amyloid beta protein and prevents amyloid formation. Proc Natl Acad Sci U S A 1994;91:8368–72.10.1073/pnas.91.18.8368Search in Google Scholar

9. Golabek AA, Soto C, Vogel T, Wisniewski T. The interaction between apolipoprotein E and Alzheimer’s amyloid beta-peptide is dependent on beta-peptide conformation. J Biol Chem 1996;271:10602–6.10.1074/jbc.271.18.10602Search in Google Scholar

10. Blennow K, Hampel H. CSF markers for incipient Alzheimer’s disease. Lancet Neurol 2003;2:605–13.10.1016/S1474-4422(03)00530-1Search in Google Scholar

11. Lewczuk P, Beck G, Esselmann H, Bruckmoser R, Zimmermann R, Fiszer M, et al. Effect of sample collection tubes on cerebrospinal fluid concentrations of tau proteins and amyloid beta peptides. Clin Chem 2006;52:332–4.10.1373/clinchem.2005.058776Search in Google Scholar PubMed

12. Perret-Liaudet A, Pelpel M, Tholance Y, Dumont B, Vanderstichele H, Zorzi W, et al. Risk of Alzheimer’s disease biological misdiagnosis linked to cerebrospinal collection tubes. J Alzheimers Dis 2012;31:13–20.10.3233/JAD-2012-120361Search in Google Scholar PubMed

13. Andreasen N, Minthon L, Davidsson P, Vanmechelen E, Vanderstichele H, Winblad B, et al. Evaluation of CSF-tau and CSF-A beta 42 as diagnostic markers for Alzheimer disease in clinical practice. Arch Neurol-Chicago 2001;58:373–9.10.1001/archneur.58.3.373Search in Google Scholar PubMed

14. Schoonenboom NSM, Mulder C, Vanderstichele H, Van Elk EJ, Kok A, Van Kamp GJ, et al. Effects of processing and storage conditions on amyloid beta (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

15. 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

16. Simonsen AH, Bahl JMC, Danborg PB, Lindstrom V, Larsen SO, Grubb A, et al. Pre-analytical factors influencing the stability of cerebrospinal fluid proteins. J Neurosci Meth 2013;215:234–40.10.1016/j.jneumeth.2013.03.011Search in Google Scholar PubMed

17. Leitao MJ, Baldeiras I, Herukka SK, Pikkarainen M, Leinonen V, Simonsen AH, et al. Chasing the effects of pre-analytical confounders – a multicenter study on CSF-AD biomarkers. Front Neurol 2015;6:153.10.3389/fneur.2015.00153Search in Google Scholar PubMed PubMed Central

18. Vanderstichele H, Van Kerschaver E, Hesse C, Davidsson P, Buyse MA, Andreasen N, et al. Standardization of measurement of beta-amyloid(1-42) in cerebrospinal fluid and plasma. Amyloid 2000;7:245–58.10.3109/13506120009146438Search in Google Scholar PubMed

19. Gordon DJ, Tappe R, Meredith SC. Design and characterization of a membrane permeable N-methyl amino acid-containing peptide that inhibits Abeta1-40 fibrillogenesis. J Pept Res 2002;60:37–55.10.1034/j.1399-3011.2002.11002.xSearch in Google Scholar PubMed

20. Soto P, Griffin MA, Shea JE. New insights into the mechanism of Alzheimer amyloid-beta fibrillogenesis inhibition by N-methylated peptides. Biophys J 2007;93:3015–25.10.1529/biophysj.107.112086Search in Google Scholar PubMed PubMed Central

21. Findeis MA. Peptide inhibitors of beta amyloid aggregation. Curr Top Med Chem 2002;2:417–23.10.2174/1568026024607508Search in Google Scholar PubMed

22. Parthsarathy V, McClean PL, Holscher C, Taylor M, Tinker C, Jones G, et al. A novel retro-inverso peptide inhibitor reduces amyloid deposition, oxidation and inflammation and stimulates neurogenesis in the APPswe/PS1DeltaE9 mouse model of Alzheimer’s disease. Plos One 2013;8:e54769.10.1371/journal.pone.0054769Search in Google Scholar PubMed PubMed Central

23. van Groen T, Wiesehan K, Funke SA, Kadish I, Nagel-Steger L, Willbold D. Reduction of Alzheimer’s disease amyloid plaque load in transgenic mice by D3, A D-enantiomeric peptide identified by mirror image phage display. ChemMedChem 2008;3:1848–52.10.1002/cmdc.200800273Search in Google Scholar PubMed

24. Amijee H, Bate C, Williams A, Virdee J, Jeggo R, Spanswick D, et al. The N-methylated peptide SEN304 powerfully inhibits A beta(1-42) toxicity by perturbing oligomer formation. Biochemistry-US 2012;51:8338–52.10.1021/bi300415vSearch in Google Scholar PubMed

25. Kokkoni N, Stott K, Amijee H, Mason JM, Doig AJ. N-Methylated peptide inhibitors of beta-amyloid aggregation and toxicity. Optimization of the inhibitor structure. Biochemistry-US 2006;45:9906–18.10.1021/bi060837sSearch in Google Scholar PubMed

26. Taylor BM, Sarver RW, Fici G, Poorman RA, Lutzke BS, Molinari A, et al. Spontaneous aggregation and cytotoxicity of the beta-amyloid A beta(1-40): a kinetic model. J Protein Chem 2003;22:31–40.10.1023/A:1023063626770Search in Google Scholar

27. McKoy AF, Chen J, Schupbach T, Hecht MH. A novel inhibitor of amyloid beta (Abeta) peptide aggregation: from high throughput screening to efficacy in an animal model of Alzheimer disease. J Biol Chem 2012;287:38992–9000.10.1074/jbc.M112.348037Search in Google Scholar PubMed PubMed Central

28. Chyan YJ, Poeggeler B, Omar RA, Chain DG, Frangione B, Ghiso J, et al. Potent neuroprotective properties against the Alzheimer beta-amyloid by an endogenous melatonin-related indole structure, indole-3-propionic acid. J Biol Chem 1999;274:21937–42.10.1074/jbc.274.31.21937Search in Google Scholar PubMed

29. Gervais F, Paquette J, Morissette C, Krzywkowski P, Yu M, Azzi M, et al. Targeting soluble Abeta peptide with Tramiprosate for the treatment of brain amyloidosis. Neurobiol Aging 2007;28:537–47.10.1016/j.neurobiolaging.2006.02.015Search in Google Scholar PubMed

30. Garman EF, Mitchell EP. Glycerol concentrations required for cryoprotection of 50 typical protein crystallization solutions. J Appl Crystallogr 1996;29:584–7.10.1107/S0021889896004190Search in Google Scholar

31. Berejnov V, Husseini NS, Alsaied OA, Thorne RE. Effects of cryoprotectant concentration and cooling rate on vitrification of aqueous solutions. J Appl Crystallogr 2006;39:244–51.10.1107/S0021889806004717Search in Google Scholar

32. Park JW, Lanier TC, Green DP. Cryoprotective effects of sugar, polyols, and or phosphates on Alaska pollack surimi. J Food Sci 1988;53:1–3.10.1111/j.1365-2621.1988.tb10163.xSearch in Google Scholar

33. Berge G, Lauridsen C, Sando SB, Holder DJ, Moller I, Aasly JO, et al. Effect of Tween-20 on core biomarkers measured in cerebrospinal fluid from patients with Alzheimer’s disease, mild cognitive impairment, or healthy control individuals. J Alzheimers Dis 2016;49:493–502.10.3233/JAD-150234Search in Google Scholar PubMed

34. Chuang VTG, Kragh-Hansen U, Otagiri M. Pharmaceutical strategies utilizing recombinant human serum albumin. Pharmaceut Res 2002;19:569–77.10.1023/A:1015396825274Search in Google Scholar

35. Bittner T, Zetterberg H, Teunissen CE, Ostlund RE, Jr., Militello M, Andreasson U, et al. Technical performance of a novel, fully automated electrochemiluminescence immunoassay for the quantitation of beta-amyloid (1-42) in human cerebrospinal fluid. Alzheimers Dement 2015;12:517–26.10.1016/j.jalz.2015.09.009Search in Google Scholar PubMed

36. McLaurin J, Franklin T, Zhang X, Deng J, Fraser PE. Interactions of Alzheimer amyloid-beta peptides with glycosaminoglycans effects on fibril nucleation and growth. Eur J Biochem 1999;266:1101–10.10.1046/j.1432-1327.1999.00957.xSearch in Google Scholar PubMed

37. Hatami A, Monjazeb S, Glabe C. The anti-amyloid-beta monoclonal antibody 4G8 recognizes a generic sequence-independent epitope associated with alpha-synuclein and islet amyloid polypeptide amyloid fibrils. J Alzheimers Dis 2016;50:517–25.10.3233/JAD-150696Search in Google Scholar PubMed

38. Walsh DM, Tseng BP, Rydel RE, Podlisny MB, Selkoe DJ. The oligomerization of amyloid beta-protein begins intracellularly in cells derived from human brain. Biochemistry-US 2000;39:10831–9.10.1021/bi001048sSearch in Google Scholar PubMed

39. Ramakrishnan M, Kandimalla KK, Wengenack TM, Howell KG, Poduslo JF. Surface plasmon resonance binding kinetics of Alzheimer’s disease amyloid beta peptide-capturing and plaque-binding monoclonal antibodies. Biochemistry-US 2009;48:10405–15.10.1021/bi900523qSearch in Google Scholar PubMed PubMed Central

40. Pica-Mendez AM, Tanen M, Dallob A, Tanaka W, Laterza OF. Nonspecific binding of A beta 42 to polypropylene tubes and the effect of Tween-20. Clin Chim Acta 2010;411:1833.10.1016/j.cca.2010.07.019Search in Google Scholar PubMed

Supplemental Material:

The online version of this article (DOI: https://doi.org/10.1515/cclm-2016-1061) offers supplementary material, available to authorized users.

Received: 2016-11-22
Accepted: 2016-12-29
Published Online: 2017-2-4
Published in Print: 2017-8-28

©2017 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2016-1061
Keywords for this article
Aβ-peptide; Alzheimer disease; cerebrospinal fluid; Elecsys β-amyloid (1-42) assay

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  2. Editorials
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