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Insulin-loaded nanoparticles were prepared by ionotropic pre-gelation of alginate with calcium chloride followed by complexation between alginate and chitosan. The influence of the pH and stoichiometry relationship between polyelectrolytes providing individual particles with a nano-scale size was assessed by photon correlation spectroscopy (PCS) and scanning electron microscopy (SEM). Insulin–polyelectrolyte interactions at varying pH and polyelectrolytes stoichiometry were assessed by differential scanning calorimetry (DSC) and Fourier-transform infrared (FTIR) studies. Individual and smaller sizing nanoparticles, around 800nm, were obtained at pH 4.7 with an alginate:chitosan mass ratio of 6:1. Thermograms of insulin-loaded nanoparticles originated shifts on same unloaded nanoparticle peaks and suggested polyelectrolytes–protein interactions at pH around 4.5–5.0. FTIR spectra of insulin-loaded …