Microparticle Deposition on Human Serum Albumin Layers: Unraveling Anomalous Adsorption Mechanism

Abstract

Human serum albumin (HSA) layers are adsorbed on mica under controlled diffusion transport at pH 3.5 and various ionic strengths. The surface concentration of HSA is directly determined by AFM imaging of single molecules. It is shown that the adsorption kinetics derived in this way is quantitatively described using the random sequential (RSA) adsorption model. The electrokinetic characteristics of the HSA layers at various pHs comprising their zeta potential are acquired in situ while using the streaming potential method. It is shown that at pH 3.5 the zeta potential of mica becomes positive for HSA concentrations above 3000 μm−2. At larger pHs, HSA layers exhibit negative zeta potential for the entire range of coverage. Thorough characteristics of these monolayers at various pHs were performed applying the colloid deposition method involving negatively charged polystyrene microparticles. The kinetics of their deposition and their maximum coverage are determined as a function of the HSA layer surface concentration, pH, and ionic strength. An anomalous deposition of microparticles on substrates also exhibiting a negative zeta potential is observed, which contradicts the Derjaguin, Landau, Vervey, Overbeek (DLVO) theory. This effect is interpreted in terms of heterogeneous charge distribution that results from molecule concentration fluctuations. It is also shown that the maximum concentration of microparticles abruptly decreases with the electric double-layer thickness that is regulated by changing ionic strength, which indicates that their deposition is governed by electrostatic interactions. One can argue that the results obtained in this work can be exploited as useful reference data for the analysis of deposition phenomena of bioparticles on protein layers.