Development of the Sequential Binding Model and Application for Anticooperative Protein Adsorption onto Charged Dendrimers.

The Langmuir binding model provides one of the simplest and elegant methods for characterizing an adsorption process. Despite its wide-ranging applications, enormous effort has been spent to further integrate complexity onto the standard Langmuir isotherm to incorporate a wide breadth of binding kinetics with the heterogeneity and cooperative effect among ligands and receptors. Here, we use statistical mechanics as a convenient theoretical framework to depict several adsorption processes on a Langmuir-like description. With regard to the system with a two-component mixture of macromolecular binders, we have derived the two-group sequential binding isotherm as an important extension of the original sequential model with more applications, including systems of non-identical binders. Via comparison of the Langmuir equilibrium with the Boltzmann equilibrium, for the first time the binding free energy defined in the Langmuir-like models can be meaningfully compared with simulations. In a practical example of the adsorption between the lysozyme protein and charged dendrimer, we have demonstrated how the calorimetry data of this system could be interpreted by the binding models described above, with an accurate description of the adsorption process, including the cooperative effect and dendrimer heterogeneity. Using the computer simulation as a benchmark, we also reveal and discuss the strengths and limitations of the proposed binding models. The entire analysis serves as a starting point for extending the standard Langmuir model to access more complicated binding processes.