Time-Resolved Optical Waveguide Spectroscopy for Studying Protein Adsorption Kinetics

Abstract

A time-resolved evanescent wave absorption technique is employed in studying the adsorption of a mammalian protein (cytochrome c) from an electrolyte-free solution on a hydrophilic glass surface. The method combines the sensitivity of a fast-scan spectral analyzer and surface specificity of slab optical waveguide (SOWG) technique. While proteins are viewed to undergo an overall irreversible adsorption process brought about by structural changes that occur after the initial adsorption phase, we have used an SOWG technique to investigate the first stage of the adsorption process while no noticeable changes have taken place. Absorbance data within the first two seconds after contact fit well into the reversible adsorption model characteristic of Langmuir-type adsorption but the goodness-of-fit diminishes as the protein molecules spend more time on the surface.