Biophys J, October 2002, p. 1965-1973, Vol. 83, No. 4

Real-Time Measurement of Spontaneous Antigen-Antibody Dissociation

Simone Kulin,^* Rani Kishore,^* Joseph B. Hubbard, and Kristian Helmerson^*

 ^*Physics Laboratory and  Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, Maryland 20899 USA

We report observations in real time of thermally driven adhesion and dissociation between a monoclonal IgE antibody and its specific antigen N--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.

Biophys J, October 2002, p. 1965-1973, Vol. 83, No. 4
© 2002 by the Biophysical Society   0006-3495/02/10/1965/09  $2.00

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