The electrospray technique was utilized to modify whey protein (WPI) and enhance its suitability for use in food products. Whey protein (3% w/w) in an aqueous/ethanol solution (47 and 50% w/w) was subjected to electrospraying under various conditions, applied voltage 10–20 kV and flow rate 0.25–1 mL/h. By controlling the flow rate and applied voltage, particles in the nano scale range were successfully produced and remained stable for a period up to 5 months. Different levels of denatured WPI were investigated by varying the alcohol content and applying heat, and their physicochemical behavior was monitored using Scanning Electron Microscopy (SEM) and Fourier-transform infrared (FTIR) spectral analysis. The impact of electrosprayed particles on cell viability was assessed on the Caco-2 cell monolayers, revealing that their effects were dose-dependent. No cytotoxic effects were observed except at higher particle concentrations, suggesting their potential safety for use.