Short-Range Interaction between Adsorbed Layers of Human Serum Albumin

Abstract

The adsorption of human serum albumin (HSA) onto muscovite mica has been investigated by means of surface force and ESCA measurements. The range of protein concentration explored was 0.001-1.0 mg/ml in 10-3M NaCl at pH 5.6. It was found that negatively charged albumin adsorbs onto negatively charged mica. Small structural changes are induced by the adsorption, and further structural changes can be induced by applying an external compressive force. The structure of the adsorbed layer depends on the surface density. As surfaces with a low adsorption density of HSA are brought together, the protein molecules are able to diffuse along the surface, facilitating the merging of the two HSA layers into one single HSA layer in the gap between the surfaces. At high surface density crowding effects impede the rearrangement of the HSA layers and two separate layers of HSA remain in the gap between the surfaces. An adhesion force is present at low-packing but not at high-packing densities. It was found that the short-range interaction between one HSA-coated surface and one bare mica surface was strongly attractive, demonstrating that the adhesion forces likely are induced by HSA molecules bridging between two surfaces and not by attractive interactions between the proteins themselves.