Fig. 5.

The effect of temperature on antibody-antigen unbinding force. a Unbinding forces of antibody-antigen interactions were measured in vitro using anti-vimentin antibody-immobilized AFM probe and vimentin-immobilized substrate. The maximum peak force detected on force curve during cantilever retraction was defined as ‘unbinding force’. b Measured unbinding forces were plotted as a histogram for the cases of 4 and 37 °C, and for negative control (no vimentin modification on the substrate). Fitting curves for the cases at 4 and 37 °C on vimentin-coated surface were obtained assuming multiple Gaussian distribution after first peak at around 80 pN, although single peak was indicated in data of 4 and 37 °C omitting the peak of nonspecific interaction at 80 pN and not clear peaks of multiple unbinding. Fitting curve for negative control data was obtained by assuming single Gaussian distribution