Measurement of Single Molecular Interactions by Dynamic Force Microscopy

  • Martin Hegner
  • Wilfried Grange
  • Patricia Bertoncini
Part of the Methods in Molecular Biology™ book series (MIMB, volume 242)


Unbinding forces of weak, noncovalent bonds have been measured by scanning force microscopy (1) or biomembrane force probes (2). Initially, these scanning force microscopy measurements focused on feasibility studies to measure single biomolecular interactions (3, 4, 5). Recently, however, a few groups showed that these single molecule experiments give a direct link to bulk experiments where thermodynamic data are experimentally acquired (6, 7, 8, 9). In contrast with bulk experiments where averaged properties are measured, a single molecular approach gives access to properties that are hidden in the ensemble. These experiments can give insight into the geometry of the energy landscape of a biomolecular bond (7,9, 10, 11). Some experiments even showed that intermediate states during unbinding (unfolding) exist which only can be detected by single molecule experiments (12, 13, 14).


Force Curve Atomic Force Microscope Cantilever Single Molecule Experiment Unbinding Force Bulk Experiment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Dammer, U., Popescu, O., Wagner, P., Anselmetti, D., Güntherodt, H. J., and Misevic G. N. (1995) Binding strength between cell-adhesion proteoglycans measured by atomic-force microscopy. Science 267, 1173–1175.PubMedCrossRefGoogle Scholar
  2. 2.
    Evans, E., and Ritchie, K. (1997) Dynamic strength of molecular adhesion bonds. Biophys. J. 72, 1541–1555.PubMedCrossRefGoogle Scholar
  3. 3.
    Moy, V. T., Florin, E. L., and Gaub, H. E. (1994) Intermolecular forces and energies between ligands and receptors. Science 266, 257–259.PubMedCrossRefGoogle Scholar
  4. 4.
    Dammer, U., Hegner, M., Anselmetti, D., et al. (1996) Specific antigen/antibody interactions measured by force microscopy. Biophys. J. 70, 2437–2441.PubMedCrossRefGoogle Scholar
  5. 5.
    Hinterdorfer, P., Baumgartner, W., Gruber, H. J., Schilcher, K., and Schindler, H. (1996) Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. Proc. Natl. Acad. Sci. USA 93, 3477–3481.PubMedCrossRefGoogle Scholar
  6. 6.
    Evans, E. (1998) Energy landscapes of biomolecular adhesion and receptor anchoring at interfaces explored with dynamic force spectroscopy. Faraday Dis. 111, 1–16.CrossRefGoogle Scholar
  7. 7.
    Strunz, T., Oroszlan, K., Schäfer, R., and Güntherodt, H. J. (1999) Dynamic force spectroscopy of single DNA molecules. Proc. Natl. Acad. Sci. USA 96, 11277–11282.PubMedCrossRefGoogle Scholar
  8. 8.
    Schwesinger, F., Ros, R., Strunz, T., et al. (2000) Unbinding forces of single antibody-antigen complexes correlate with their thermal dissociation rates. Proc. Natl. Acad. Sci. USA 97, 9972–9977.PubMedCrossRefGoogle Scholar
  9. 9.
    Schumakovitch, I., Grange, W., Strunz, T., Bertoncini, P., Güntherodt, H.-J., and Hegner, M. (2002) Temperature dependence of unbinding forces between complementary DNA strands. Biophys. J. 82, 517–521.PubMedCrossRefGoogle Scholar
  10. 10.
    Evans, E. (2001) Probing the relation between force-lifetime-and chemistry in single molecular bonds. Annu. Rev. Biophys. Biomol. Struct. 30, 105–128.PubMedCrossRefGoogle Scholar
  11. 11.
    Merkel, R. (2001) Force spectroscopy on single passive biomolecules and single biomolecular bonds. Phys. Rep. 346, 343–385.CrossRefGoogle Scholar
  12. 12.
    Merkel, R., Nassoy, P., Leung, A., Ritchie, K., and Evans, E. (1999) Energy landscape of receptor-ligand bonds explored with dynamic force spectroscopy. Nature 397, 50–53.PubMedCrossRefGoogle Scholar
  13. 13.
    Di Paris, R., Strunz, T., Oroszlan, K., Güntherodt, H.-J., and Hegner, M. (2000) Dynamics of molecular complexes under an applied force. Single Mol. 1, 285–290.CrossRefGoogle Scholar
  14. 14.
    Marszalek, P. E., Lu, H., Li, H. B., al. (1999) Mechanical unfolding intermediates in titin modules. Nature 402, 100–103.PubMedCrossRefGoogle Scholar
  15. 15.
    Strunz, T., Oroszlan, K., Schumakovitch, I., Güntherodt, H.-J., and Hegner, M. (2000) Model energy landscapes and the force-induced dissociation of ligand-receptor bonds. Biophys. J. 79, 1206–1212.PubMedCrossRefGoogle Scholar
  16. 16.
    Schwarzenbach, M. S., Reimann, P., Thommen, V., et al. (2002) Interferon α-2a interactions on glass vial surfaces measured by atomic force microscopy. PDA J. Pharm. Sci. Technol. 56, 78–89.PubMedGoogle Scholar
  17. 17.
    Hutter J. L. and Bechhoefer, J. (1993) Calibration of atomic-force micro-scopetips. Rev. Sci. Instrum. 64, 3342–3342.CrossRefGoogle Scholar
  18. 18.
    Izenman, A. J. (1991) Recent developments in nonparametric density estimation. J. Am. Stat. Assoc. 86, 205–224.CrossRefGoogle Scholar
  19. 19.
    Hermanson, G. T. (1996) Bioconjugate Techniques, Academic Press, San Diego.Google Scholar
  20. 20.
    Hegner M. (2000) DNA handles for single molecule experiments. Single Mol. 1, 139–144.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Martin Hegner
    • 1
  • Wilfried Grange
    • 1
  • Patricia Bertoncini
    • 2
  1. 1.NCCR Nanoscale Science, Department of PhysicsUniversity of BaselBaselSwitzerland
  2. 2.Department of Physics, NCCR Nanoscale ScienceUniversity of BaselBaselSwitzerland

Personalised recommendations