The adsorption of bovine serum albumin (BSA) in a planar poly(acrylic acid) (PAA) brush layer has been studied by fixed-angle optical reflectometry. The influence of polymer length, grafting density, and salt concentration is studied as a function of pH. The results are compared with predictions of an analytical polyelectrolyte brush model, which incorporates charge regulation and excluded volume interactions. A maximum in adsorption is found near the point of zero charge (pzc) of the protein. At the maximum, BSA accumulates in a PAA brush to at least 30 vol %. Substantial adsorption continues above the pzc, that is, in the pH range where a net negatively charged protein adsorbs into a negatively charged brush layer, up to a critical pH value. This critical pH value decreases with increasing ionic strength. The adsorbed amount increases strongly with both increasing PAA chain length and increasing grafting density. Experimental data compare well with the analytical model without having to include a nonhomogeneous charge distribution on the protein surface. Instead, charge regulation, which implies that the protein adjusts its charge due to the negative electrostatic potential in the brush, plays an important role in the interpretation of the adsorbed amounts. Together with nonelectrostatic interactions, it explains the significant protein adsorption above the pzc.

Colloidal probe atomic force microscopy was used to determine the electric double layer interactions between a gold electrode and a spherical silica probe. The double layer properties of the gold/solution interface were varied through the pH and salt concentration of the electrolyte, as well as by externally applying an electric potential. The double layer potentials ψ(d) of the gold surface were obtained by fitting the force-distance curves according to the DLVO (Derjaguin-Landau-Verwey-Overbeek) theory, using earlier obtained values for the double layer potential of the silica probe as input parameter. It was found that the gold electrode combines the features of reversible and polarizable interfaces; i.e., its charge and potential are determined by both the solution pH and the external potential. The pH dependence is attributed to proton adsorption and desorption from oxidic groups on the gold surface. In the potential range studied, ψ(d) varies linearly with the applied potential; the variation in ψ(d) is roughly 10% of that in the applied potential. The potential of zero force (the external potential at which ψ(d) = 0) varies with pH. The various features of the gold/electrolyte interface are described well by an amphifunctional double layer model. The results of this study form the basis of the interpretation of adsorption studies on gold as a function of pH and externally applied potential.

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Protein encapsulation with polymers has a high potential for drug delivery, enzyme protection and stabilization. Formation of such structures can be achieved by the use of polyelectrolytes to generate so-called complex coacervate core micelles (C3Ms). Here, encapsulation of enhanced green fluorescent protein (EGFP) was investigated using a cationic-neutral diblock copolymer of two different sizes: poly(2-methyl-vinyl-pyridinium)41-b-poly(ethylene-oxide)205 and poly(2-methyl-vinyl-pyridinium)128-b-poly(ethylene-oxide)477. Dynamic light scattering and fluorescence correlation spectroscopy (FCS) revealed a preferred micellar composition (PMC) with a positive charge composition of 0.65 for both diblock copolymers and micellar hydrodynamic radii of approximately 34 nm. FCS data show that at the PMC, C3Ms are formed above 100 nM EGFP, independent of polymer length. Mixtures of EGFP and nonfluorescent GFP were used to quantify the amount of GFP molecules per C3M, resulting in approximately 450 GFPs encapsulated per micelle. This study shows that FCS can be successfully applied for the characterization of protein-containing C3Ms.