Self-assembly of Nafion onto in situ formed silica nanoparticles in ethylene glycol-water mixture solvent has been investigated in this study. It was found that the formation of silica nanoparticles depends on the concentration of Nafion in dispersions. At relatively low concentration, 0.8% in weight in this case, the existing Nafion is not sufficient to prevent further growth of the initially formed silica nanoparticles, leading to large aggregates of silica particles. When the concentration of Nafion increased to 2% in weight, self-assembled Nafion layer on the surface stabilizes the initial formed silica nanoparticles and silica particles with average diameters of 4.2+/-0.5 nm were found to be uniformly distributed in the dispersion. With further increasing the concentration of Nafion, the number of Nafion aggregates increases and silica nanoparticles were mainly formed inside the entangled Nafion chains, resulting in an observation of clusters of silica nanoparticles.