Microchimica Acta

, 186:117 | Cite as

A photoelectrochemical sandwich immunoassay for protein S100β, a biomarker for Alzheimer’s disease, using an ITO electrode modified with a reduced graphene oxide-gold conjugate and CdS-labeled secondary antibody

  • Mahmoud Amouzadeh TabriziEmail author
  • Josep Ferré-Borrull
  • Pankaj Kapruwan
  • Lluis F. MarsalEmail author
Original Paper


A sandwich-type photoelectrochemical immunoassay is described for the protein S100ß which is an Alzheimer’s disease biomarker found in the astrocytes of the brain. Antibody against S100ß (anti-S100ß) was labeled with CdS quantum dots and then acted as a secondary antibody. The labeled antibody was characterized by FTIR, ultraviolet-visible and fluorescence spectroscopy. An indium-tin oxide (ITO) electrode was modified with a nanocomposite prepared from reduced graphene oxide and gold nanoparticles. Then, a sol-gel film containing isocyanate functional groups (-N=C=O) was cast on the surface of the electrode. The NCO group reacts with amino groups of the labeled antibody to covalently bind them to the surface. The S100β was bound by the primary immobilized antibody on the rGO-Au/ITO electrode and then sandwiched with the labeled secondary antibody. Cyclic voltammetry and electrochemical impedance spectroscopy were applied to confirm the stepwise changes in the electrochemical properties of the electrode surface. The photoelectrochemical immunoassay, typically operated at a potential of +0.2 V (vs. Ag|AgClsat) gives a signal that is related to the logarithm of the S100β concentration in the range from 0.25 to 10 ng·mL−1 with a lower detection limit of 0.15 pg·mL−1. The method was successfully applied to the determination of S100β in human serum samples.

Graphical abstract

Schematic presentation of an immunosensor which is based on an indium tin oxide modified with reduced graphene oxide decorated with gold nanocomposite and antibody. The immunosensor was applied for the determination of S100β biomarker by using in the labeled antibody.


Alzheimer’ disease S100β biomarker Photoelectrochemical immunoassay Reduced graphene oxide-gold conjugate CdS-labeled secondary antibody 



This study was supported by the Spanish Ministry of Economy and Competition under grants number TEC2015–71324-R (MINECO/FEDER), the Catalan Government AGAUR 2017-SGR-1527, the ICREA under the 2014-ICREA Academia Award and the Martí-Franquès II postdoctoral programme under grant number 2017PMF-POST2-7. The research leading to these results has received funding from” la Caixa bank” Foundation.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3159_MOESM1_ESM.docx (3.4 mb)
ESM 1 (DOCX 3.36 mb)


  1. 1.
    Chaves ML, Camozzato AL, Ferreira ED, Piazenski I, Kochhann R, Dall'Igna O, Mazzini GS, Souza DO, Portela LV (2010) Serum levels of S100B and NSE proteins in Alzheimer's disease patients. J Neuroinflammation 7:6–6CrossRefGoogle Scholar
  2. 2.
    Felix FS, Angnes L (2018) Electrochemical immunosensors – a powerful tool for analytical applications. Biosens Bioelectron 102:470–478CrossRefGoogle Scholar
  3. 3.
    Rajeev G, Xifre-Perez E, Prieto Simon B, Cowin AJ, Marsal LF, Voelcker NH (2018) A label-free optical biosensor based on nanoporous anodic alumina for tumour necrosis factor-alpha detection in chronic wounds. Sensors Actuators B Chem 257:116–123CrossRefGoogle Scholar
  4. 4.
    Liu L, Zhou X, Wilkinson JS, Hua P, Song B, Shi H (2017) Integrated optical waveguide-based fluorescent immunosensor for fast and sensitive detection of microcystin-LR in lakes: optimization and analysis. Sci Rep 7:3655–3664CrossRefGoogle Scholar
  5. 5.
    Della Ventura B, Sakač N, Funari R, Velotta R (2017) Flexible immunosensor for the detection of salivary α-amylase in body fluids. Talanta 174:52–58CrossRefGoogle Scholar
  6. 6.
    Crosson C, Rossi C (2013) Quartz crystal microbalance immunosensor for the quantification of immunoglobulin G in bovinemilk. Biosens Bioelectron 42:453–459CrossRefGoogle Scholar
  7. 7.
    Kokkinos C, Economou A, Prodromidis MI (2016) Electrochemical immunosensors: critical survey of different architectures and transduction strategies. TrAC Trends Anal Chem 79:88–105CrossRefGoogle Scholar
  8. 8.
    Eissa S, L'Hocine L, Siaj M, Zourob M (2013) A graphene-based label-free voltammetric immunosensor for sensitive detection of the egg allergen ovalbumin. Analyst 138:4378–4384CrossRefGoogle Scholar
  9. 9.
    Meng L, Gan N, Li T, Cao Y, Hu F, Zheng L (2011) A three-dimensional, magnetic and electroactive Nanoprobe for Amperometric determination of tumor biomarkers. Int J Mol Sci 12:362–375CrossRefGoogle Scholar
  10. 10.
    Jiang W, Wu L, Duan J, Yin H, Ai S (2018) Ultrasensitive electrochemiluminescence immunosensor for 5-hydroxymethylcytosine detection based on Fe3O4@SiO2 nanoparticles and PAMAM dendrimers. Biosens Bioelectron 99:660–666CrossRefGoogle Scholar
  11. 11.
    Wu L, Li M, Zhang M, Yan M, Ge S, Yu J (2013) Ultrasensitive electrochemiluminescence immunosensor for tumor marker detection based on nanoporous sliver@carbon dots as labels. Sensors Actuators B Chem 186:761–767CrossRefGoogle Scholar
  12. 12.
    Zhou C, Liu D, Xu L, Li Q, Song J, Xu S, Xing R, Song H (2015) A sensitive label–free amperometric immunosensor for alpha-fetoprotein based on gold nanorods with different aspect ratio. Sci Rep 5:9939–9946CrossRefGoogle Scholar
  13. 13.
    Amouzadeh Tabrizi M, Shamsipur M, Mostafaie A (2016) A high sensitive label-free immunosensor for the determination of human serum IgG using overoxidized polypyrrole decorated with gold nanoparticle modified electrode. Mater Sci Eng C 59:965–969CrossRefGoogle Scholar
  14. 14.
    Wang H, Wang Y, Zhang Y, Wang Q, Ren X, Wu D, Wei Q (2016) Photoelectrochemical Immunosensor for detection of carcinoembryonic antigen based on 2D TiO2 Nanosheets and Carboxylated graphitic carbon nitride. Sci Rep 6:27385–27392CrossRefGoogle Scholar
  15. 15.
    Qin C, Bai X, Zhang Y, Gao K (2018) Photoelectrochemical CdSe/TiO2 nanotube array microsensor for high-resolution in-situ detection of dopamine. Microchim Acta 185:278CrossRefGoogle Scholar
  16. 16.
    Liu X, Wei C, Luo J, Wu Y, Guo X, Ying Y, Wen Y, Yang H (2018) Photoelectrochemical determination of the activity of M.SssI methyltransferase, and a method for inhibitor screening. Microchim Acta 185:498CrossRefGoogle Scholar
  17. 17.
    Lin Y, Zhou Q, Zeng Y, Tang D (2018) Liposome-coated mesoporous silica nanoparticles loaded with L-cysteine for photoelectrochemical immunoassay of aflatoxin B1. Microchim Acta 185:311CrossRefGoogle Scholar
  18. 18.
    Liu X-P, Xie X-L, Wei Y-P, C-j M, Chen J-S, Niu H-L, Song J-M, Jin B-K (2017) Photoelectrochemical immunoassay for human interleukin 6 based on the use of perovskite-type LaFeO3 nanoparticles on fluorine-doped tin oxide glass. Microchim Acta 185:52CrossRefGoogle Scholar
  19. 19.
    Zhao W-W, Wang J, Zhu Y-C, Xu J-J, Chen H-Y (2015) Quantum dots: Electrochemiluminescent and Photoelectrochemical bioanalysis. Anal Chem 87:9520–9531CrossRefGoogle Scholar
  20. 20.
    Wang X, Yan T, Li Y, Liu Y, Du B, Ma H, Wei Q (2015) A competitive photoelectrochemical immunosensor based on a CdS-induced signal amplification strategy for the ultrasensitive detection of dexamethasone. Sci Rep 5:17945–17953CrossRefGoogle Scholar
  21. 21.
    Cui R, Pan H-C, Zhu J-J, Chen H-Y (2007) Versatile Immunosensor using CdTe quantum dots as electrochemical and fluorescent labels. Anal Chem 79:8494–8501CrossRefGoogle Scholar
  22. 22.
    Amouzadeh Tabrizi M, Tavakkoli A, Dhand V, Rhee KY, Park S-J (2014) Eco-friendly one-pot synthesis of gold decorated reduced graphene oxide using beer as a reducing agent. J Ind Eng Chem 20:4327–4331CrossRefGoogle Scholar
  23. 23.
    Wang G-L, Jiao H-J, Zhu X-Y, Dong Y-M, Li Z-J (2012) Enhanced fluorescence sensing of melamine based on thioglycolic acid-capped CdS quantum dots. Talanta 93:398–403CrossRefGoogle Scholar
  24. 24.
    Elevathoor Vikraman A, Rosin Jose A, Jacob M, Girish Kumar K (2015) Thioglycolic acid capped CdS quantum dots as a fluorescent probe for the nanomolar determination of dopamine. Anal Methods 7:6791–6798CrossRefGoogle Scholar
  25. 25.
    Pang L, Cui H, Liu Y, Zhong W (2016) Anti-VEGF antibody conjugated CdHgTe quantum dots as a fluorescent probe for imaging in living mouse. J Lumin 173:274–278CrossRefGoogle Scholar
  26. 26.
    Yang H, Guo Q, He R, Li D, Zhang X, Bao C, Hu H, Cui D (2009) A quick and parallel analytical method based on quantum dots labeling for ToRCH-related antibodies. Nanoscale Res Lett 4:1469–1474CrossRefGoogle Scholar
  27. 27.
    Xing S-G, Xiong Q-R, Zhong Q, Zhang Y, Bian S-M, Jin Y, Chu X-G (2013) Recent research advances of antibody-conjugated quantum dots. Chin J Anal Chem 41:949–954CrossRefGoogle Scholar
  28. 28.
    Saunders AE, Popov I, Banin U (2006) Synthesis of hybrid CdS−au colloidal nanostructures. J Phys Chem B 110:25421–25429CrossRefGoogle Scholar
  29. 29.
    Harrison MT, Kershaw SV, Burt MG, Eychmüller A, Weller H, Rogach AL (2000) Wet chemical synthesis and spectroscopic study of CdHgTe nanocrystals with strong near-infrared luminescence. Mater Sci Eng B 69-70:355–360CrossRefGoogle Scholar
  30. 30.
    Khetani S, Aburashed R, Singh A, Sen A, Sanati-Nezhad A (2017) Immunosensing of S100β biomarker for diagnosis of spinal cord injuries (SCI). Sensors Actuators B Chem 247:163–169CrossRefGoogle Scholar
  31. 31.
    Liu Y, Wang H, Chen J, Liu C, Li W, Kong J, Yang P, Liu B (2013) A sensitive microchip-based Immunosensor for electrochemical detection of low-level biomarker S100B. Electroanalysis 25:1050–1055CrossRefGoogle Scholar
  32. 32.
    Souada M, Piro B, Reisberg S, Anquetin G, Noël V, Pham MC (2015) Label-free electrochemical detection of prostate-specific antigen based on nucleic acid aptamer. Biosens Bioelectron 68:49–54CrossRefGoogle Scholar
  33. 33.
    Kurzątkowska K, Jankowska A, Wysłouch-Cieszyńska A, Zhukova L, Puchalska M, Dehaen W, Radecka H, Radecki J (2016) Voltammetric detection of the S100B protein using his-tagged RAGE domain immobilized onto a gold electrode modified with a dipyrromethene–cu(II) complex and different diluents. J Electroanal Chem 767:76–83CrossRefGoogle Scholar
  34. 34.
    Mikuła E, Wysłouch-Cieszyńska A, Zhukova L, Puchalska M, Verwilst P, Dehaen W, Radecki J, Radecka H (2014) Voltammetric detection of S100B protein using his-tagged receptor domains for advanced glycation end products (RAGE) immobilized onto a gold electrode surface. Sensors (Basel, Switz) 14:10650–10663CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Departamento de Ingeniería Electrónica, Eléctrica y AutomáticaUniversitat Rovira i VirgiliTarragonaSpain

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