Protein adsorption from flowing solutions on pure and maleic acid copolymer modified glass particles

Abstract

The adsorption of human serum albumin (HSA) and lysozyme (LSZ) on pure as well as maleic acid (MA) copolymer coated spherical soda lime glass particles was investigated under flowing conditions. Coating the glass particles with two different maleic acid copolymers alters the properties of the particle surface concerning its charge and hydrophobicity in a well-defined gradation. Frontal chromatography was used to determine the surface concentration of the adsorbed proteins and to establish adsorption isotherms. The introduced methodology was demonstrated to provide a powerful means to study protein adsorption at solid/liquid interfaces. Investigations with virginal and protein-preadsorbed glass particles revealed that even under streaming conditions HSA is irreversibly adsorbed, whereas LSZ partially desorbs. For LSZ and HSA the adsorbed amounts and the isotherms strongly depend on the surface “history”, i.e. the presence or absence of preadsorbed protein layers, and the kind of surface modification of the glass. Compared to the soda lime glass surface the adsorption of HSA was strongly increased on surfaces modified with a hydrophobic maleic acid copolymer indicating a strong hydrophobic protein–surface interaction. By coating the surface with a hydrophilic and more negatively charged maleic acid copolymer the adsorption of HSA to that surface was lower and comparable to the adsorption onto plain glass due to the electrostatic repulsion between HSA and the modified surface. In contrast the affinity to any of the investigated particle surfaces was generally higher for LSZ than for HSA which can be mainly attributed to the electrostatic attraction between LZS and the surface. The adsorbed amount of LSZ on the copolymer coated particle surfaces was much higher than on the pure soda lime glass particles indicating superposed hydrophobic interactions in the case of the hydrophobic MA copolymer layer and an increased density of anionic sites as well as interactions of LSZ within the three-dimensional (swollen), hydrophilic MA copolymer layer.