Abstract
We report a free-solution, label-free method for quantitative characterization of macromolecular interactions using dynamic light scattering, a temperature controlled plate reader, and a multiwell concentration gradient. This nondestructive technique enabled determination of stoichiometry of binding, equilibrium dissociation constant, and thermodynamic parameters, as well as the impact of temperature, buffer salinity, and a small-molecule inhibitor. The low volume capability of dynamic light scattering reduced the required sample to 426 pmol/experiment, with detection limits for 150-kDa proteins anticipated to be in the low femtomole range.