Real-time measurement of spontaneous antigen-antibody dissociation

Biophys J. 2002 Oct;83(4):1965-73. doi: 10.1016/S0006-3495(02)73958-1.

Abstract

We report observations in real time of thermally driven adhesion and dissociation between a monoclonal IgE antibody and its specific antigen N-epsilon-2,4-dinitrophenyl-L-lysine. Both molecules were attached to the surfaces of different polystyrene microspheres trapped by optical tweezers. Monitoring spontaneous successive attachment and detachment events allowed a direct determination of the reaction-limited detachment rate k(off) for a single bond and also for multiple bonds. We observed both positive and negative cooperativity between multiple bonds depending on whether the antigen was linked to the microsphere with or without a tether, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigen-Antibody Reactions*
  • Biophysical Phenomena
  • Biophysics
  • Cell Adhesion
  • Dose-Response Relationship, Drug
  • Flow Cytometry
  • Haptens / chemistry
  • Immunoglobulin E / chemistry*
  • Immunoglobulin E / metabolism
  • Kinetics
  • Lysine / analogs & derivatives*
  • Lysine / chemistry*
  • Lysine / metabolism
  • Time Factors

Substances

  • Haptens
  • epsilon-dinitrophenyllysine
  • Immunoglobulin E
  • Lysine