β-conglycinin nanoparticles were prepared by self-assembly with desolvation method. The effect of protein concentration, volume ratio of ethanol to protein solution and amount of glutaraldehyde on nano-size was studied using turbidity method, and the release behavior of Vc embedded in the nanoparticles was determined. The results showed that the nanoparticles had better dispersity when the protein concentration, the volume ratio of ethanol to protein solution and the amount of glutaraldehyde (1.5 g/L) was 10-12 g/L, 4:1 and 0.5 mL, respectively. The nano-size was mainly around 350-380 nm. Vc in crosslinked nanoparticles released smoothly while Vc in uncrosslinked nanoparticles had a sudden accelerated phenomenon in the release rate, and the release mechanism of Vc in pH3.0 and 7.4 buffer solutions was Fickian diffusion and anomalous transport, respectively.