Protein adsorption: kinetics and history dependence.

The adsorption of protein and other macromolecular species is often strongly history dependent owing to the slow relaxation of nonequilibrium structures. The rate at which molecules adsorb is exquisitely sensitive to, and thus a sensitive measure of, interfacial structure; adsorption kinetics may therefore serve both to identify and to quantify history dependence. Optical waveguide lightmode spectroscopy (OWLS) in multistep mode, where an adsorbing surface is alternately exposed to a protein solution and one free of protein, enables the comparison of adsorption rates onto interfacial layers of identical composition but different formation histories. Contributions to the overall kinetics from the apparent adsorption rate constant, the desorption rate constants of molecules in different adsorbed states, and the interfacial one-body cavity function (a measure of the probability of a "cavity" on the surface free of protein) may be isolated from OWLS kinetic data. We find the rate of adsorption and the corresponding cavity function to increase significantly-for systems of fibronectin, cytochrome c, and lysozyme adsorbing onto Si(Ti)O(2)-from a first to a subsequent adsorption step and attribute this to a clustering transition among adsorbed protein molecules. From the kinetic data, we determine important structural and temporal information on this event.