Analytical and Bioanalytical Chemistry

, Volume 408, Issue 14, pp 3675–3680 | Cite as

Nanocrystalline diamond sensor targeted for selective CRP detection: an ATR-FTIR spectroscopy study

  • Per Ola AnderssonEmail author
  • Pernilla Viberg
  • Pontus Forsberg
  • Fredrik Nikolajeff
  • Lars Österlund
  • Mikael Karlsson
Rapid Communication


Protein immobilization on functionalized fluorine-terminated nanocrystalline (NCD) films was studied by attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy using an immobilization protocol developed to specifically bind C-reactive protein (CRP). Using an ATR-FTIR spectroscopy method employing a force-controlled anvil-type configuration, three critical steps of the ex situ CRP immobilization were analyzed. First, the NCD surface was passivated by deposition of a copolymer layer consisting of polyethylene oxide and polypropylene oxide. Second, a synthetic modified polypeptide binder with high affinity to CRP was covalently attached to the polymeric film. Third, CRP dissolved in aqueous buffer in concentrations of 10–20 μg/mL was added on the functionalized NCD surface. Both the amide I and II bands, due to the polypeptide binder and CRP, were clearly observed in ATR-FTIR spectra. CRP amide I bands were extracted from difference spectra and yielded bands that agreed well with the reported amide I band of free (non-bonded) CRP in solution. Thus, our results show that CRP retains its secondary structure when it is attached to the polypeptide binders. Compared to previous IR studies of CRP in solution, about 200 times lower concentration was applied in the present study.

Graphical Abstract

Direct non-destructive ATR-FTIR analysis of C-reactive protein (CRP) selectively bound to functionalized nanocrystalline diamond (NCD) sensor surface


Infrared spectroscopy ATR-FTIR Nanocrystalline diamond CRP Protein binders Biosensor 



Dr. Karin Fromell and Prof. Lars Baltzer (both at Uppsala University) are acknowledged for their help with binder and CRP samples. The work was supported financially by grants from Uppsala Berzelii Technology Centre for Neurodiagnostics and VR-project 621-2014-5959.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Per Ola Andersson
    • 1
    • 2
    • 3
    Email author
  • Pernilla Viberg
    • 2
  • Pontus Forsberg
    • 2
  • Fredrik Nikolajeff
    • 2
    • 3
  • Lars Österlund
    • 2
    • 3
  • Mikael Karlsson
    • 2
    • 3
  1. 1.CBRN Defence and SecurityFOI Swedish Defence Research AgencyUmeåSweden
  2. 2.Department of Engineering SciencesUppsala UniversityUppsalaSweden
  3. 3.Molecular Fingerprint Sweden ABUppsalaSweden

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