Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Protein-modified nanocrystalline diamond thin films for biosensor applications

Nature Materials volume 3, pages 736–742 (2004)Cite this article

Abstract

Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: AFM and KFM images of a patterned NCD sample.
Figure 2: Summary of the biofunctionalization process.
Figure 3: XPS spectra of the biofunctionalized NCD samples.
Figure 4: Fluorescence image of a NCD sample patterned by oxidation and functionalized with GFP.
Figure 5: Cyclic voltammograms (CV), showing the i–U current–voltage curves of the electrodes.
Figure 6: Cyclic voltammograms of an untreated NCD electrode (black) and a catalase-modified NCD electrode (red), measured in pure phosphate buffer.

Similar content being viewed by others

References

  1. Collings, A.F. & Caruso, F. Biosensors: recent advances. Rep. Prog. Phys. 60, 1397–1445 (1997).

    Article  CAS  Google Scholar 

  2. Bergveld, P. Development of an ion-sensitive solid-state device for neurophysiological measurements. IEEE Trans. Biomed. Eng. BME 17, 70–72 (1970).

    Article  Google Scholar 

  3. Linford, M.R., Fenter, P., Eisenberger, P.M. & Chidsey, C.E.D. Alkyl monolayers on silicon prepared from 1-alkenes and hydrogenated silicon. J. Am. Chem. Soc. 117, 3145–3155 (1995).

    Article  CAS  Google Scholar 

  4. Cai, W. et al. Chemical modification and patterning of iodine-terminated silicon surfaces using visible light. J. Phys. Chem. B 106, 2656–2664 (2002).

    Article  CAS  Google Scholar 

  5. Lin, Z. et al. DNA attachment and hybridization at the silicon (100) surface. Langmuir 18, 788–796 (2002).

    Article  CAS  Google Scholar 

  6. Bakowicz, K. & Mitura, S. Biocompatibility of NCD. J. Wide Bandgap Mater. 9, 261–272 (2002).

    Article  CAS  Google Scholar 

  7. Miller, J.B. & Brown, D.W. Photochemical modification of diamond films. Langmuir 12, 5809–5817 (1996).

    Article  CAS  Google Scholar 

  8. Ando, T. et al. Diffuse reflectance infrared Fourier-transform study of the direct thermal fluorination of diamond powder surfaces. J. Chem. Soc. Faraday Trans. 91, 3209–3212 (1995).

    Article  CAS  Google Scholar 

  9. Ohtani, B. et al. Surface functionalization of doped CVD diamond via covalent bond. An XPS study on the formation of surface-bound quaternary pyridinium salt. Chem. Lett. 27, 953–954 (1998).

    Article  Google Scholar 

  10. Gruen, D.M. Nanocrystalline diamond films. Annu. Rev. Mater. Sci. 29, 211–259 (1999).

    Article  CAS  Google Scholar 

  11. Teukam, Z. et al. Shallow donors with high n-type electrical conductivity in homoepitaxial deuterated boron-doped diamond layers. Nature Mater. 2, 482–486 (2003).

    Article  CAS  Google Scholar 

  12. Koizumi, S., Watanabe, K., Hasegawa, M. & Kanda, H. Ultraviolet emission from a diamond pn junction. Science 292, 1899–1901 (2001).

    Article  CAS  Google Scholar 

  13. Hupert, M. et al. Conductive diamond thin-films in electrochemistry. Diamond Rel. Mater. 12, 1940–1949 (2003).

    Article  CAS  Google Scholar 

  14. Strother, T. et al. Photochemical functionalization of diamond films. Langmuir 18, 968–971 (2002).

    Article  CAS  Google Scholar 

  15. Yang, W. et al. DNA-modified nanocrystalline diamond thin-films as stable, biologically active substrates. Nature Mater. 1, 253–257 (2002).

    Article  CAS  Google Scholar 

  16. Pleskov, Y.U., Sakharova, A.Y., Krotova, M.D., Bouilov, L.L. & Spitsyn, B.V. Photoelectrochemical properties of semiconductor diamond. J. Electroanal. Chem. 228, 19–27 (1987).

    Article  CAS  Google Scholar 

  17. Landstrass, M.I. & Ravi, K.V. Resistivity of chemical vapor deposited diamond films. Appl. Phys. Lett. 55, 975–977 (1989).

    Article  CAS  Google Scholar 

  18. Maier, F., Riedel, M., Mantel, B., Ristein. J. & Ley, L. Origin of surface conductivity in diamond. Phys. Rev. Lett. 85, 3472–3475 (2000).

    Article  CAS  Google Scholar 

  19. Garrido, J.A., Nebel, C.E. & Stutzmann, M. Capacitance–voltage studies of Al-Schottky contacts on hydrogen-terminated diamond. Appl. Phys. Lett. 81, 637–639 (2002).

    Article  CAS  Google Scholar 

  20. Birrell, J. et al. Bonding structure in nitrogen doped ultrananocrystalline diamond. J. Appl. Phys. 93, 5606–5612 (2003).

    Article  CAS  Google Scholar 

  21. Rezek, B. et al. Fermi level on hydrogen terminated diamond surfaces. Appl. Phys. Lett. 82, 2266–2268 (2003).

    Article  CAS  Google Scholar 

  22. Chalfie, M., Tu, Y., Euskirchen, G., Ward, W.W. & Prasher, D.C. Green fluorescent protein as a marker for gene-expression. Science 263, 802–805 (1994).

    Article  CAS  Google Scholar 

  23. Armstrong, F.A. & Wilson, G.S. Recent developments in faradaic bioelectrochemistry. Electrochim. Acta 45, 2623–2645 (2000).

    Article  CAS  Google Scholar 

  24. Lai, M.E. & Bergel, A. Direct electrochemistry of catalase on glassy carbon electrodes. Bioelectrochemistry 55, 157–160 (2002).

    Article  CAS  Google Scholar 

  25. Zhang, Z., Chouchane, S., Magliozzo, R.S. & Rusling, J.F. Direct voltammetry and catalysis with mycobacterium tuberculosis catalase-peroxidase, peroxidases, and catalase in lipid films. Anal. Chem. 74, 163–170 (2002).

    Article  CAS  Google Scholar 

  26. Switala, J. & Loewen, P.C. Diversity of properties among catalases. Arch. Biochem. Biophys. 401, 145–154 (2002).

    Article  CAS  Google Scholar 

  27. Schoenfisch, M.H. & Pemberton, J.E. Air stability of alkanethiol self-assembled monolayers on silver and gold surfaces. J. Am. Chem. Soc. 120, 4502–4513 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank M. Haselbeck for providing the green fluorescent protein, and E. Bertel and N. Memmel for discussions on the properties of nanocrystalline diamond, as well as the Raman spectra shown as supplementary information. We also thank the staff of BESSY for help during the experiments. A.H. acknowledges the Technische Universität München for financial support within a PhD fellowship. J.H. thanks the Alexander von Humboldt Foundation for financial support.

Author information

Authors and Affiliations

  1. Walter Schottky Institut, Technische Universität München, Am Coulombwall 3, Garching, 85748, Germany

    Andreas Härtl, Evelyn Schmich, Jose A. Garrido, Jorge Hernando & Martin Stutzmann

  2. Institut für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, Garching, 85748, Germany

    Silvia C. R. Catharino & Stefan Walter

  3. Physics Department E20, Technische Universität München, James-Franck-Strasse, Garching, 85748, Germany

    Peter Feulner

  4. r-Best coating, Hartstoffbeschichtungs GmbH, Erlach 165, Steinach, 6150, Austria

    Alexander Kromka & Doris Steinmüller

Authors
  1. Andreas Härtl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Evelyn Schmich
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jose A. Garrido
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jorge Hernando
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Silvia C. R. Catharino
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Stefan Walter
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Peter Feulner
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Alexander Kromka
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Doris Steinmüller
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Martin Stutzmann
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jose A. Garrido.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Härtl, A., Schmich, E., Garrido, J. et al. Protein-modified nanocrystalline diamond thin films for biosensor applications. Nature Mater 3, 736–742 (2004). https://doi.org/10.1038/nmat1204

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nmat1204

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing