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.
![](http://media.springernature.com/lw685/springer-static/image/art%3A10.1007%2Fs00604-014-1180-2/MediaObjects/604_2014_1180_Figa_HTML.gif)
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.
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs00604-014-1180-2/MediaObjects/604_2014_1180_Fig1_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs00604-014-1180-2/MediaObjects/604_2014_1180_Fig2_HTML.gif)
![](http://media.springernature.com/m312/springer-static/image/art%3A10.1007%2Fs00604-014-1180-2/MediaObjects/604_2014_1180_Fig3_HTML.gif)
References
Jellinger KA (2003) Neropathological spectrum of synucleinopathies. Mov Disord 18:2–12
Eller M, Williams DR (2011) α-synuclein in Parkinson disease and other neurodegenerative disorders. Clin Chem Lab Med 49:403–408
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
Corresponding authors
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
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00604-014-1180-2