Analytical and Bioanalytical Chemistry

, Volume 391, Issue 5, pp 1861–1869

Effect of the immobilisation of DNA aptamers on the detection of thrombin by means of surface plasmon resonance

  • Veronika Ostatná
  • Hana Vaisocherová
  • Jiří Homola
  • Tibor Hianik
Original Paper

DOI: 10.1007/s00216-008-2133-6

Cite this article as:
Ostatná, V., Vaisocherová, H., Homola, J. et al. Anal Bioanal Chem (2008) 391: 1861. doi:10.1007/s00216-008-2133-6
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Abstract

We report a multichannel surface plasmon resonance (SPR) sensor for detection of thrombin via DNA aptamers immobilized on the SPR sensor surface. A detailed investigation of the effect of the immobilisation method on the interaction between thrombin and DNA aptamers is presented. Three basic approaches to the immobilisation of aptamers on the surface of the SPR sensor are examined: (i) immobilisation based on chemisorption of aptamers modified with SH groups, (ii) immobilisation of biotin-tagged aptamers via previously immobilized avidin, neutravidin or streptavidin molecular linkers, and (iii) immobilisation employing dendrimers as a support layer for subsequent immobilisation of aptamers. A level of nonspecific binding of thrombin to immobilized human serum albumin (HSA) for each of the immobilisation methods is determined. Immobilisation of aptamers by means of the streptavidin–biotin system yields the best results both in terms of sensor specificity and sensitivity.

Keywords

DNA aptamerThrombinDendrimersSurface plasmon resonance

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Veronika Ostatná
    • 1
  • Hana Vaisocherová
    • 2
  • Jiří Homola
    • 2
  • Tibor Hianik
    • 1
  1. 1.Department of Nuclear Physics and BiophysicsComenius UniversityBratislavaSlovak Republic
  2. 2.Institute of Photonics and ElectronicsAcademy of Sciences of the Czech RepublicPragueCzech Republic