Log in

A porous silicon immunoassay platform for fluorometric determination of α-synuclein in human cerebrospinal fluid

  • Short Communication
  • Published:
Microchimica Acta Aims and scope Submit manuscript

Abstract

Levels of total and/or oligomeric α-synuclein may be used as a biomarker tool to aid in the diagnosis and development of new disease-modifying therapies. We report here on a porous silicon antibody microarray for the fluorimetric determination of cerebrospinal fluid levels of total α-synuclein, a protein involved the pathology of Parkinson’s disease. The surface of porous silicon has a 3-dimensional macro- and nanoporous structure, and this offers a large binding capacity for capturing probe molecules. Porous silicon also warrants efficient immobilization of antibodies by surface adsorption, and does not require chemical immobilization. The platform requires 10 μL of cerebrospinal fluid, and each test requires 4 h for assay only (including immobilization of capturing antibody). The limit of detection is 35 pg mL−1 of α-synuclein in cerebrospinal fluid, and the dynamic analytical range extends from 0.01 to 100 ng·mL−1.

High antibody capturing capacity of porous silicon allows high density of antibody immobilization on the surface and make it possible enriching binding event to target protein (α−synuclein). Below shows SEM images of porous silicon surface and assayed microarray images.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (France)

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

References

  1. Jellinger KA (2003) Neropathological spectrum of synucleinopathies. Mov Disord 18:2–12

    Article  Google Scholar 

  2. Eller M, Williams DR (2011) α-synuclein in Parkinson disease and other neurodegenerative disorders. Clin Chem Lab Med 49:403–408

    Article  CAS  Google Scholar 

  3. Mollenhauer B, Locascio JJ, Schulz-Schaeffer W, Sixel-Döring F, Trenkwalder C, Schlossmacher MG (2011) α-synuclein and tau concentrations in cerebrospinal fluid of patients presenting with parkinsonism: a cohort study. Lancet Neurol 10:230–240

    Article  CAS  Google Scholar 

  4. Hall S, Öhrfelt A, Constantinescu R, Andreasson U, Surova Y, Bostrom F, Nilsson C, Håkan W, Decraemer H, Någga K, Minthon L, Londos E, Vanmechelen E, Holmberg B, Zetterberg H, Blennow K, Hansson O (2012) Accuracy of a panel of 5 cerebrospinal fluid biomarkers in the differential diagnosis of patients with dementia and/or parkinsonian disorders. Arch Neurol 69:1445–1452

    Article  Google Scholar 

  5. Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, Pike B, Root H, Rubenstein J, Boyer R, Stenroos ES, Chandrasekharappa S, Athanassiadou A, Papapetropoulos T, Johnson WG, Lazzarini AM, Duvoisin RC, Di Iorio G, Golbe LI, Nussbaum RL (1997) Mutation in the alpha-synuclein gene identified in families with Parkinson’s disease. Science 276:2045–2047

    Article  CAS  Google Scholar 

  6. Kruger R, Kuhn W, Muller T, Woitalla D, Graeber M, Kosel S, Przuntek H, Epplen JT, Schols L, Riess O (1998) Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson’s disease. Nat Genet 18:106–108

    Article  CAS  Google Scholar 

  7. Zarranz J, Alegre J, Gomez-Esteban JC, Lezcano E, Ros R, Ampuero I, Vidal L, Hoenicka J, Rodriguez O, Atares B, Llorens V, Gomez Tortosa E, del Ser T, Munoz DG, de Yebenes JG (2004) The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 55:164–173

    Article  CAS  Google Scholar 

  8. Singleton AB, Farrer M, Johnson J, Singleton A, Hague S, Kachergus J, Hulihan M, Peuralinna T, Dutra A, Nussbaum R, Lincoln S, Crawley A, Hanson M, Maraganore D, Adler C, Cookson MR, Muenter M, Baptista M, Miller D, Blancato J, Hardy J, Gwinn-Hardy K (2003) Alpha-Synuclein locus triplication causes Parkinson’s disease. Science 302:841

    Article  CAS  Google Scholar 

  9. Chartier-Harlin MC, Kachergus J, Roumier C, Mouroux V, Douay X, Lincoln S, Levecque C, Larvor L, Andrieux J, Hulihan M, Waucquier N, Defebvre L, Amouyel P, Farrer M, Destee A (2004) Alpha-synuclein locus duplication as a cause of familial Parkinson’s disease. Lancet 364:1167–1169

    Article  CAS  Google Scholar 

  10. El-Agnaf OM, Salem, Paleologou KE, Cooper LJ, Fullwood NJ, Gibson MJ (2003) Alpha-synuclein implicated in Parkinson’s disease is present in extracellular biological fluids, including human plasma. FASEB J 17:1945–1947

    CAS  Google Scholar 

  11. Tokoda T, Qureshi MM, Ardah MT, Varghese S, Shehab SA, Kasai T (2010) Detection of elevated levels of alpha-synuclein in cerebrospinal oligomers in CSF from patients with Parkinson disease. Neurology 75:1766–1772

    Article  Google Scholar 

  12. Steinhauer C, Ressine A, Marko-Varga G, Laurell, Borrebaeck C, Wingren C (2005) Biocompatibility of surfaces for antibody microarrays: design of macroporous silicon substrates. Anal Biochem 341:204–213

    Article  CAS  Google Scholar 

  13. Järås K, Ressine A, Nilsson E, Malm J, Marko-Varga G, Lilja H, Laurell T (2007) Reverse versus sandwich antibody microarrays – a comparison from a clinical perspective. Anal Chem 79:5817–5825

    Article  Google Scholar 

  14. Järås K, Adler B, Lilja H, Malm J, Marko-Varga G, Laurell T (2012) PSA quantitation of 80 plasma samples from the clinical routine using antibody microarrays. Clin Chim Acta 414:76–84

    Article  Google Scholar 

  15. Lee SW, Kim S, Malm J, Jeong OC, Lilja H, Laurell T (2013) Improved porous silicon microarray based prostate specific antigen immunoassay by optimized surface density of the capture antibody. Anal Chim Acta 796:108–114

    Article  CAS  Google Scholar 

  16. Hong Z, Shi M, Chung K, Quinn J, Peskind E, Glasko D, Jankovic J, Zabetian C, Leverenz J, Baird G, Montine T, Hancock A, Hwang H, Pan C, Brandner J, Kang U, Jensen P, Zhang J (2010) DJ-1 and α-synuclein in human cerebrospinal fluid as biomarkers of Parkinson’s disease. Brain 133:713–726

    Article  Google Scholar 

  17. Ressine A, Marko-Varga G, Laurell T (2007) Porous silicon protein microarray technology and ultra- superhydrophobic states for improved bioanalytical readout. Biotechnol Annu Rev 13:149–200

    Article  CAS  Google Scholar 

  18. Tokuda T, Salem S, Allsop D, Mizuno T, Nakagawa M, Qureshi M, Locascio JJ, Schlossmacher MG, El-Agnaf O (2006) Decreased α–synuclein in cerebrospinal fluid of aged individuals and subjects with Parkinson’s disease. Biochem Biophys Res Commun 349:162–166

    Article  CAS  Google Scholar 

  19. Wood WG (2008) Immunoassays & co.: past, present, future-A review and outlook from personal experience and involvement over the past 35 years. Clin Lab 54:423–438

    CAS  Google Scholar 

  20. Mollenhauer B, Cullen V, Kahn I, Krastins B et al (2008) Direct quantification of CSF α-synuclein by ELISA and first cross-sectional study in patients with neurodegeneration. Exp Neurol 213:315–325

    Article  CAS  Google Scholar 

  21. Aerts MB, Esselink R, Bloem BR, Verbeek MM (2011) Cerebrospinal fluid tau and phosphorylated tau protein are elevated in corticobasal syndrome. Mov Dis 26:169–173

    Article  CAS  Google Scholar 

  22. Tinsley RB, Kotschet K, Modesto D, Ng H, Wang Y, Nagley P, Shaw G, Horne MK (2010) Sensitive and specific detection of α-synuclein in human plasma. J Neurosci Res 88:2693–2700

    CAS  Google Scholar 

Download references

Acknowledgment

The Swedish Research Council-Linnaeus Grant, Bagadilico, The Swedish Research Council (grant no. 621-2009-5361; 2009-5361) and Korea- Swedish Research Cooporation Program (STINT) and STINT Institutional Grant: IG2010 2068. This project also supported by Korea Ministry of Environment as “EI project” (ERL E211-41003-0007-0), and Agency for Defence Development through Chemical and Biological Defence Research Center (2012-0126-005).

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Sangwook Lee or Thomas Laurell.

Additional information

Sangwook Lee and Edina Silajdžić equally contributed to the paper.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 763 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lee, S., Silajdžić, E., Yang, H. et al. A porous silicon immunoassay platform for fluorometric determination of α-synuclein in human cerebrospinal fluid. Microchim Acta 181, 1143–1149 (2014). https://doi.org/10.1007/s00604-014-1180-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00604-014-1180-2

Keywords

Navigation