The mechanism of separation methods, for example, liquid chromatography, is realized through rapid multiple adsorption‐desorption steps leading to the dynamic equilibrium state in a mixture of molecules with different partition coefficients. Sorting of colloidal particles, including protein complexes, cells, and viruses, is limited due to a high energy barrier, up to millions kT, required to detach particles from the interface, which is in dramatic contrast to a few kT for small molecules. Such a strong interaction renders particle adsorption quasi‐irreversible. The dynamic adsorption‐desorption equilibrium is approached very slowly, if ever attainable. This limitation is alleviated with a local oscillating repulsive mechanical force generated at the microstructured stimuli‐responsive polymer interface to switch between adsorption and mechanical‐force‐facilitated desorption of the particles. Such a dynamic regime enables the separation of colloidal mixtures based on the particle‐polymer interface affinity, and it could find use in research, diagnostics, and industrial‐scale label‐free sorting of highly asymmetric mixtures of colloids and cells.