In this study, we compare the preparation of ovalbumin (OVA) and α-lactalbumin (α-LA) nanoparticles using different desolvating agents (ethanol, acetone, and methanol) and water: desolvating agent volume ratios (1:3, 1:4, 1:5, 1:10, and 1:20). Also the effects of protein solution temperature (25, 50, and 80 ℃) on the size of nanoparticles and the stability of crosslinked nanoparticles for 30 d were studied. OVA and α-LA were shown to be good candidates for nanoparticulation and nanoparticles in the range of 60 to 230 nm were obtained. The comparison between the 2 proteins offers guidance to optimize OVA and α-LA nanoparticle fabrication and to efficiently obtain nanoparticles with desired characteristics. The particle sizes of OVA nanoparticles were found to be in the range of 60 to 160 nm, and the particle sizes of α-LA were between 150 and 230 nm. The sizes varied with different desolvating agents: for OVA, ethanol, and methanol both produced nanoparticles smaller than 100 nm; for α-LA, methanol produced the smallest nanoparticles. Water: desolvating agent ratios, in the studied range, did not show a significant effect on the particle sizes for both OVA and α-LA nanoparticles. The size and morphology of the nanoparticles were found to change when the protein solutions were heated up to 50 and 80 ℃ and cooled down before nanoparticulation and most nanoparticles had a smaller diameter.