Abstract: In order to improve the soluble VP2 protein expression of Porcine parvovirus（PPV） and to develop a vaccine with virus-like particles（VLPs）, in the experiment, after codon optimization, the PPV VP2 gene sequence was synthesized and pET30a-PPV-VP2 recombinant expression plasmid was constructed, and it was co-transformed with three molecular chaperones, pKJE7, pGro7 and pTf16, into E. coli BL21（DE3） competent cells. After induction, the soluble expression level was analyzed and the induction conditions were optimized, and SDS-PAGE and Western-blot were performed for identification. Purified by Ni-NTA affinity chromatography, imidazole was removed by dialysis for in vitro self-assembly, and the particle size and morphology of VLPs were observed by dynamic light scattering and transmission electron microscopy. The immunogenicity of the prepared PPV VLPs vaccine was evaluated by using intramuscular immunization and nasal drip immunization of Kunming rats. The results showed that when pET30a-PPV-VP2 was co-expressed with pTf16, the soluble expression of VP2 protein in E. coli was the highest when 0.1 mmol/L IPTG, 2 g/L l-arabinose, and 30 ℃ incubation for 16 h were used as the induction conditions. The VP2 protein was purified to self-assemble into PPV VLPs with a diameter of approximately 23.52 nm and a hemagglutination potency of 1∶2~9. The expressed VLPs were prepared as a vaccine and immunized by intramuscular injection in Kunming mice; the HI titer was highly significantly higher than that of the inactivated Porcine parvovirus vaccine（P＜0.001）, and the mice were induced to produce higher levels of antibodies（IgG1, IgG2a, IgG2b, IgG3） and cytokines（γ-interferon IFN-γ and interleukin-4 IL-4）. Nasal droplet immunization with VLPs induced the production of high levels of secretory immunoglobulin（sIgA）, and the VLPs had no significant side effects and were safe. The results indicated that the prepared PPV VLPs had good immunogenicity, induced cellular and humoral immunity, balanced Th1/Th2 immune response and efficient mucosal immune response in Kunming rats without adjuvant-assisted nasal drop VLPs vaccine.