Enzyme Adsorption on Nanoparticle Surface Probed by Highly Sensitive Second Harmonic Light Scattering

Recent developments in second harmonic light scattering technique and the associated theoretical models have provided a deeper insight of molecular interactions on micro- and nanoparticle surfaces. This technique is extended to probe the thermodynamics of protein adsorption on nanoparticle surface which is crucial for understanding the fate of nanoparticle-based formulations in biomedical applications. A modified Langmuir adsorption model has been applied to extract the thermodynamic parameters from the experimental data. The general applicability of the technique is established by extracting free energy change, association constant, and binding stoichiometry of adsorption of a moderate size protein, alcohol dehydrogenase, and a small size protein, insulin, on gold nanoparticles. The free energy change for the adsorption is found to be of the order of -55kJ/mol, which indicates that the interaction of proteins with the nanoparticle surface involves weak forces. On the other hand, the low value of the free energy change makes the detachment of the protein from the particle surface easier and guarantees reversibility of the binding process. In addition, one gets the binding stoichiometry of the proteins with the nanoparticle surface which opens up the possibility of controlling the payload of the protein- or peptide-based therapeutics in future biomedical applications.