Thermodynamic analysis of r-hGH-polymer surface Interaction using isothermal titration calorimetry.

Delivery of protein drugs would be an effective approach if mechanistic understanding of protein adsorption at solid/liquid surface is obtained and nonspecific adsorption can be controlled. This study involves evaluation of the thermodynamic parameters for interaction between recombinant human growth hormone (r-hGH) and nanoparticles of poly (lactic- co- glycolic) acid polymer of different molecular weight. Energy changes related to protein-nanoparticle interactions are usually very small and isothermal titration calorimetry (ITC) is the only technique that determines the binding constant (Ka), enthalpy, (ΔH) and stoichiometry, (n) in a single experiment. Therefore, we used ITC to study the energetics of the binding of r-hGH with PLGA nanoparticles. The largely negative ∆H and ∆S suggested that binding was driven by non-classical hydrophobic effect for interaction with PLGA 50501A and PLGA 8515E nanoparticles at pH 7.2 where higher surface coverage was noted for the latter. Endothermic, entropically driven reaction was observed upon interaction of r-hGH with PLGA 5050 5E nanoparticles at pH 7.2, PLGA 5050 1A nanoparticles at pH 5.3 and pH 4.0. Moreover, negative value of ΔCp for PLGA 50501A NPs at pH 7.2 indicated cooperative disorder of hydrogen-bonding networks and no evidence of hydrophobic elements. ITC proved to be very efficient method in studying the thermodynamics of the protein polymer interaction. In agreement with results from previous studies using fluorescence spectroscopy, circular dichroism and dynamic light scattering in our laboratory, this study demonstrated that adsorption can be controlled by selecting the polymer of low to moderate hydrophobicity depending on the pH of media.