Antibody-unfolding and metastable-state binding in Force spectroscopy and recognition imaging

Parminder Kaur, Qiang-Fu, Alexander Fuhrmann, Robert Ros, Linda Obenauer Kutner, Lumelle A. Schneeweis, Ryman Navoa, Kirby Steger, Lei Xie, Christopher Yonan, Ralph Abraham, Michael J. Grace, Stuart Lindsay

Research output: Contribution to journalArticlepeer-review

16 Scopus citations


Force spectroscopy and recognition imaging are important techniques for characterizing and mapping molecular interactions. In both cases, an antibody is pulled away from its target in times that are much less than the normal residence time of the antibody on its target. The distribution of pulling lengths in force spectroscopy shows the development of additional peaks at high loading rates, indicating that part of the antibody frequently unfolds. This propensity to unfold is reversible, indicating that exposure to high loading rates induces a structural transition to a metastable state. Weakened interactions of the antibody in this metastable state could account for reduced specificity in recognition imaging where the loading rates are always high. The much weaker interaction between the partially unfolded antibody and target, while still specific (as shown by control experiments), results in unbinding on millisecond timescales, giving rise to rapid switching noise in the recognition images. At the lower loading rates used in force spectroscopy, we still find discrepancies between the binding kinetics determined by force spectroscopy and those determined by surface plasmon resonance-possibly a consequence of the short tethers used in recognition imaging. Recognition imaging is nonetheless a powerful tool for interpreting complex atomic force microscopy images, so long as specificity is calibrated in situ, and not inferred from equilibrium binding kinetics.

Original languageEnglish (US)
Pages (from-to)243-250
Number of pages8
JournalBiophysical journal
Issue number1
StatePublished - Jan 5 2011

ASJC Scopus subject areas

  • Biophysics


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