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Nanobiosensors for In Vitro and In Vivo Analysis of Biomolecules

  • J.-Pablo Salvador
  • Mark P. Kreuzer
  • Romain Quidant
  • Gonçal Badenes
  • M.-Pilar Marco
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 811)

Abstract

This chapter presents as a proof of concept the development of a nanosensor based on the localized surface plasmon resonance for the analysis of biomolecules. The method presented take advantage of the plasmon generated in the surrounding of gold nanoparticles (i.e., 100 nm) for the specific interaction between antigen and antibody. The procedure for the optimization of an assay for the determination of biomolecules consisted mainly of four steps. First, the immobilization of gold nanoparticles over the glass surface using the appropriate ratio, concentration and time-contact of amino-sylilating agent, and nonreactive sylilating agent. Next, the suitable concentration of coating antigen in order to obtain the maximum signal LSPR. Following this step, the interaction between antigen and antibody (specific antibody) is evaluated by measuring the signal LSPR. Finally, a calibration curve was obtained for the detection of a small organic molecule such as stanozolol using this nanobiosensor. As a proof of concept, the use of a model is performed that in this case is for the detection of an anabolic androgenic steroid, such as stanozolol which is banned for the European Commission (EC) as a growth promoter and for the World Anti-Doping Agency (WADA) as a doping agent. The nanosensor developed demonstrates its feasibility for screening purposes due to the limit of detection achieved (0.7 μg/L) is under the MRPL required for both organizations (10 μg/L). A protocol such as that presented here may be generally applied for the analysis of other pollutant such as pesticides or antibiotics, or for biomedical applications for the analysis of biomarkers using the LSPR principle using gold nanoparticles (i.e., 30–120 nm).

Key words

Antibodies Plasmon LSPR Biosensor Stanozolol Gold nanoparticles 

Notes

Acknowledgments

This work also has been supported by the Ministry of Science and Education (contract number DEP2007-73224-C03-01). The AMR group is a consolidated Grup de Recerca de la Generalitat de Catalunya and has support from the Departament d’Universitats, Recerca i Societat de la Informació la Generalitat de Catalunya (expedient 2009 SGR 1343). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008-2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • J.-Pablo Salvador
    • 1
    • 2
  • Mark P. Kreuzer
    • 3
  • Romain Quidant
    • 3
  • Gonçal Badenes
    • 3
  • M.-Pilar Marco
    • 1
    • 2
  1. 1.CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)BarcelonaSpain
  2. 2.Applied Molecular Receptors GroupIQAC-CSICBarcelonaSpain
  3. 3.ICFO Institute of Photonic ScienceCastelldefelsSpain

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