Determination of Graphene Oxide Adsorption Space by Lysozyme Uptake—Mechanistic Studies

[Image: see text] The interaction between graphene oxide (GO) and lysozyme (LYZ) in aqueous solution was investigated for GO specific surface area determination and for the thermodynamic description of the process. It was experimentally proved that LYZ is a much better adsorbate than the most common methylene blue, allowing the determination of genuine GO surface area. Our fluorescence spectroscopy results indicate that LYZ molecules interact with GO at high- and low-affinity sites depending on the surface coverage, reflecting the protein mono- and multilayer formation, respectively. The lack of the secondary structure changes confirms LYZ usability as a model adsorbate. The calculated values of thermodynamic parameters (Δ(ΔH(0)) = −195.0 kJ/mol and Δ(ΔS(0)) = −621.3 J/molK) indicate that the interactions are exothermic, enthalpy-driven. All the reported results reveal the physical nature of the LYZ–GO interaction at the studied concentration ratios.