Fabrication and Characterization of Bioactive Thiol-Silicate Nanoparticles

  • Frances NevilleEmail author
  • Paul Millner
Part of the Methods in Molecular Biology book series (MIMB, volume 743)


Here we describe a new method for the production of thiol-silicate particles and the entrapment of enzymes within the thiol particles as they are formed. When bio-inspired polymers (polyethyleneimine) are combined with a silicic acid source and phosphate buffer under pH neutral conditions, formation of silicate particles occurs. In the method presented here the silica source contains a thiol group and so therefore the silicate particles are pre-functionalized with thiol groups. We have termed the silicate particles produced “thiol particles” and the characterization of these thiol particles is also presented in this chapter. As enzymes can be entrapped during fabrication, it means that the thiol particles can not only attach to metal surfaces but also catalyse certain reactions depending on the enzyme used. This means that there are many future possibilities for the use of thiol particles containing enzymes, as they may be used in a wide range of processes and devices which require catalytic functionalized surfaces, such as biosensors and biocatalytic reactors.

Key words

Silicates thiol nanoparticles biomimetic silica polyethyleneimine biosensors biocatalysis 



The authors would like to acknowledge the sponsorship of the SANTS project by the European Commission (Project No. NMP4-CT-2006-033254).


  1. 1.
    Niemeyer, C. M. (2001) Nanoparticles, proteins, and nucleic acids: Biotechnology meets materials science. Angew. Chem. Int. Ed. 40, 4128–4158.CrossRefGoogle Scholar
  2. 2.
    Lopez, P. J., Gautier, C., Livage, J., and Coradin, T. (2005) Mimicking biogenic silica nanostructures formation. Curr. Nanosci. 1, 73–83.CrossRefGoogle Scholar
  3. 3.
    Jin, R. H., and Yuan, J. J. (2005) Simple synthesis of hierarchically structured silicas by poly(ethyleneimine) aggregates pre-organized by media modulation. Macromol. Chem. Phys. 206, 2160–2170.CrossRefGoogle Scholar
  4. 4.
    Berne, C., Betancor, L., Luckarift, H. R., and Spain, J. C. (2006) Application of a microfluidic reactor for screening cancer prodrug activation using silica-immobilized nitrobenzene nitroreductase. Biomacromolecules 7, 2631–2636.CrossRefGoogle Scholar
  5. 5.
    He, P., Greenway, G., and Haswell, S. J. (2008) The on-line synthesis of enzyme functionalized silica nanoparticles in a microfluidic reactor using polyethylenimine polymer and R5 peptide. Nanotechnology 19, 315603.CrossRefGoogle Scholar
  6. 6.
    Helmecke, O., Hirsch, A., Behrens, P., and Menzel, H. (2008) Influence of polymeric additives on biomimetic silica deposition on patterned microstructures. J. Colloid Interface Sci. 321, 44–51.CrossRefGoogle Scholar
  7. 7.
    Betancor, L., and Luckarift, H. R. (2008) Bioinspired enzyme encapsulation for biocatalysis. Trends Biotechnol. 26, 566–572.CrossRefGoogle Scholar
  8. 8.
    Betancor, L., Luckarift, H. R., Seo, J. H., Brand, O., and Spain, J. C. (2008) Three-dimensional immobilization of β-galactosidase on a silicon surface. Biotechnol. Bioeng. 99, 261–267.CrossRefGoogle Scholar
  9. 9.
    Stöber, W., Fink, A., and Bohn, E. (1968) Controlled growth of monodisperse silica spheres in the micron size range. J. Colloid Interface Sci. 26, 62–69.CrossRefGoogle Scholar
  10. 10.
    Hernandez, G., and Rodriquez, R. (1999) Adsorption properties of silica sols modified with thiol groups. J. Non-Cryst. Solids 246, 209–215.CrossRefGoogle Scholar
  11. 11.
    Lee, Y. G., Park, J. H., Oh, C., Oh, S. G., and Kim, Y. C. (2007) Preparation of highly monodispersed hybrid silica spheres using a one-step sol-gel reaction in aqueous solution. Langmuir 23, 10875–10878.CrossRefGoogle Scholar
  12. 12.
    Pchelintsev, N. A., and Millner, P. A. (2008) A novel procedure for rapid surface functionalisation and mediator loading of screen-printed carbon electrodes. Anal. Chim. Acta 612, 190–197.CrossRefGoogle Scholar
  13. 13.
    Neville, F., Pchelintsev, N. A., Broderick, M. J. F., Gibson, T., and Millner, P. A. (2009) Novel one-pot synthesis and characterization of bioactive thiol-silicate nanoparticles for biocatalytic and biosensor applications. Nanotechnology 20, 055612.CrossRefGoogle Scholar
  14. 14.
    Ellman, G. L. (1958) A calorimetric method for determining low concentrations of mercaptans. Arch. Biochem. Biophys. 74, 443–450.CrossRefGoogle Scholar
  15. 15.
    Nakroshis, P., Amoroso, M., Legere, J., and Smith, C. (2003) Measuring Boltzmann’s constant using video microscopy of Brownian motion. Am. J. Phys. 71, 568–573.CrossRefGoogle Scholar
  16. 16.
    Carr, B., and Malloy, A. (2006) Nanoparticle tracking analysis – the NANOSIGHT system.
  17. 17.
    Roberts, E., and Rouser, G. (1958) Spectrophotometric assay for reaction of N-ethylmaleimide with sulfhydryl groups. Anal. Chem. 30, 1291–1292.CrossRefGoogle Scholar
  18. 18.
    Alexander, N. M. (1958) Spectrophotometric assay for sulfhydryl groups using N-ethylmaleimide. Anal. Chem. 30, 1292–1294.CrossRefGoogle Scholar
  19. 19.
    Nishiyama, J., and Kuninori, T. (1992) Assay of thiols and disulfides based on the reversibility of N-ethylmaleimide alkylation of thiols combined with electrolysis. Anal. Biochem. 200, 230.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.School of Engineering, University of NewcastleCallaghanAustralia
  2. 2.Institute of Membrane and Systems Biology, University of LeedsLeedsUK

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