Advertisement

Analytical and Bioanalytical Chemistry

, Volume 404, Issue 6–7, pp 1643–1651 | Cite as

Designed protein binders in combination with nanocrystalline diamond for use in high-sensitivity biosensors

  • Karin FromellEmail author
  • Pontus Forsberg
  • Mikael Karlsson
  • Karin Larsson
  • Fredrik Nikolajeff
  • Lars Baltzer
Original Paper

Abstract

A platform for diagnostic applications showing signal-to-noise ratios that by far surpass those of traditional bioanalytical test formats has been developed. It combines the properties of modified nanocrystalline diamond (NCD) surfaces and those of polyethylene oxide and polypropylene oxide based block copolymers for surface passivation and binder conjugation with a new class of synthetic binders for proteins. The NCD surfaces were fluorine-, hydrogen-, or oxygen-terminated prior to further biofunctionalization and the surface composition was characterized by X-ray photoelectron spectroscopy. In a proof of principle demonstration targeting the C-reactive protein, an ELISA carried out using an F-terminated diamond surface showed a signal-to-noise ratio of 3,900 which compares well to the signal-to-noise of 89 obtained in an antibody-based ELISA on a polystyrene microtiter plate, a standard test format used in most life science laboratories today. The increase in signal-to-noise ratio is to a large extent the result of extremely efficient passivation of the diamond surface. The results suggest that significant improvements can be obtained in standardized test formats using new materials in combination with new types of chemical coatings and receptor molecules.

Keywords

Protein binders Nanocrystalline diamond CRP Biosensor Surface characterization 

Notes

Acknowledgments

This work has enjoyed financial support from the Uppsala Berzelii Technology Centre for Neurodiagnostics and by The Swedish Diamond Centre. We also wish to thank Allvivo Inc. for supporting us with Pluronic F108-PDS.

Supplementary material

216_2012_6245_MOESM1_ESM.pdf (32 kb)
ESM 1 (PDF 69 kb)

References

  1. 1.
    Härtl A, Schmich E, Garrido JA, Hernando J, Catharino SCR, Walter S, Feulner P, Kromka A, Steinmüller D, Stutzmann M (2004) Protein-modified nanocrystalline diamond thin films for biosensor applications. Nat Mater 3:736–742CrossRefGoogle Scholar
  2. 2.
    Baltzer L (2012) Crossing borders to bind proteins—a new concept in protein recognition based on the conjugation of small organic molecules or short peptides to polypeptides from a designed set. Anal Bioanal Chem 400(6):1653–1664CrossRefGoogle Scholar
  3. 3.
    Rifai N, Ridker PM (2001) High-sensitivity C-reactive protein: a novel and promising marker of coronary heart disease. Clin Chem 47:403–411Google Scholar
  4. 4.
    Pepys MB, Hirschfield GM (2003) C-reactive protein: a critical update. J Clin Invest 111:1805–1812Google Scholar
  5. 5.
    Tegler LT, Nonglaton G, Büttner F, Caldwell K, Christopeit T, Danielsson H, Fromell K, Gossas T, Larsson A, Longati P, Norberg T, Ramapanicker R, Rydberg J, Baltzer L (2011) Efficient protein binders for the C-reactive protein from a designed chemically modified peptide library—a new concept for biomolecular recognition. Angew Chem 50:1823–1827CrossRefGoogle Scholar
  6. 6.
    Li JT, Carlsson J, Lin JN, Caldwell KD (1996) Chemical modification of surface active poly(ethylene oxide)-poly(propylene oxide) triblock copolymers. Bioconj Chem 7:592–599CrossRefGoogle Scholar
  7. 7.
    Vivensang C, Turban G, Anger E, Giquel A (1994) Reactive ion etching of diamond and diamond-like carbon films. Diam Relat Mater 3:645–649CrossRefGoogle Scholar
  8. 8.
    Shirafuji J, Sakamoto Y, Furukawa A, Shigeta H, Sugino T (1995) X-ray photoelectron spectroscopy analysis of plasma-treated surfaces of diamond films. Diam Relat Mater 4:984–988CrossRefGoogle Scholar
  9. 9.
    Maier F, Graupner R, Hollering M, Hammer L, Ristein J, Ley L (1999) The hydrogenated and bare diamond (110) surface: a combined LEED-, XPS-, and ARPES study. Surf Sci 443:177–185CrossRefGoogle Scholar
  10. 10.
    Francz G, Reinke P, Oelhafen P, Hänni W (1995) Photoelectron spectroscopy of ion-irradiated B-doped CVD diamond surfaces. Thin Solid Films 270:200–204CrossRefGoogle Scholar
  11. 11.
    Steinmüller-Nethl D, Kloss FR, Najam-Ul-Haq M, Rainer M, Larsson K, Linsmeier C, Köhler G, Gassner R, Fehrer C, Lepperdinger G, Huck CW, Bonn G (2006) Strong physisorption of BMP-2 on nanocrystalline diamond retains bioactivity. Biomaterials 27:4547–4556CrossRefGoogle Scholar
  12. 12.
    Petrini D, Larsson K (2007) A theoretical approach to the energetic stability and geometry of hydrogen and oxygen terminated diamond (100) surfaces. J Phys Chem C 111:795CrossRefGoogle Scholar
  13. 13.
    Christopeit T, Gossas T, Danielson UH (2009) Characterization of Ca2+ and phosphocholine interactions with C-reactive protein using a surface plasmon resonance biosensor. Anal Biochem 391:39–44CrossRefGoogle Scholar
  14. 14.
    Neff JA, Caldwell KD, Tresco PA (1998) A novel method for surface modification to promote cell attachment to hydrophobic substrates. J Biomed Mater Res 40:511–519CrossRefGoogle Scholar
  15. 15.
    Karlsson M, Forsberg P, Nikolajeff F (2010) From hydrophilic to superhydrophobic: fabrication of micrometer-sized nail-head-shaped pillars in diamond. Langmuir 26:889–892CrossRefGoogle Scholar
  16. 16.
    ter Veen R, Fromell K, Caldwell KD (2005) Shifts in polystyrene particle surface charge upon adsorption of the Pluronic F108 surfactant. J Colloids Interface Sci 288:124–128CrossRefGoogle Scholar
  17. 17.
    Albrecht C, Fechner P, Honcharenko D, Baltzer L, Gauglitz G (2010) A new assay design for clinical diagnostics based on alternative recognition elements. Biosens Bioelectron 25:2302–2308CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Karin Fromell
    • 1
    Email author
  • Pontus Forsberg
    • 2
  • Mikael Karlsson
    • 2
  • Karin Larsson
    • 3
  • Fredrik Nikolajeff
    • 2
  • Lars Baltzer
    • 1
  1. 1.Department of Biochemistry and Organic ChemistryUppsala UniversityUppsalaSweden
  2. 2.Department of Engineering SciencesUppsala UniversityUppsalaSweden
  3. 3.Department of Materials ChemistryUppsala UniversityUppsalaSweden

Personalised recommendations