Guo Wei, Xie Qianqian, Liu Ruipeng, Dong Hu, Zhang Yun, Wang Xiaoqiang, Sun Shiqi, Guo Huichen, Teng Zhidong
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine and Biosafety, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730000, Gansu, China.
College of Biological and Pharmaceutical Engineering, Lanzhou Jiaotong University, Lanzhou 730070, Gansu, China.
Sheng Wu Gong Cheng Xue Bao. 2025 Jul 25;41(7):2682-2693. doi: 10.13345/j.cjb.250156.
Vaccination is a crucial strategy for the prevention and control of infectious diseases. Virus-like particles (VLPs), composed of structural proteins, have garnered significant attention as a novel type of vaccine due to their excellent safety and immunogenicity. However, similar to most vaccine antigens, VLPs exhibit insufficient thermal stability, which not only restricts the widespread application of vaccines but also increases the risk of vaccine inactivation. This study aims to enhance the stability and shelf life of VLPs derived from type A foot-and-mouth disease virus (FMDV) by employing vacuum freeze-drying technology. The optimal lyoprotectant formulation was determined through single-factor and combinatorial screening. Subsequently, the correlation between the immunogenicity of the freeze-dried vaccine and the content of FMDV VLPs was evaluated via a mouse model. The stability of FMDV VLPs before and after freeze-drying was further assessed by storing them at 4, 25, and 37 ℃ for varying time periods. Results indicated that the lyoprotectant formulation No.1, composed of 7.5% trehalose, 0.1% Tween 80, 50 mmol/L glycine, 1% sodium glutamate, and 3% polyvinylpyrrolidone (PVP), effectively preserved the content of FMDV VLPs during the vacuum freeze-drying process. The immunization trial in mice revealed that the levels of specific antibodies, immunoglobulin G1 (IgG1), interleukin-4 (IL-4), and neutralizing antibodies induced by freeze-dried FMDV VLPs were comparable to those induced by non-freeze-dried FMDV VLPs. The heat treatment results showed that the storage periods of freeze-dried FMDV VLPs at 4, 25, and 37 ℃ were significantly longer than those of non-freeze-dried FMDV VLPs. In conclusion, the selected lyoprotectant formulation effectively improved the stability of FMDV VLPs vaccines. This study provides valuable insights for enhancing the stability of novel subunit vaccines.
疫苗接种是预防和控制传染病的关键策略。由结构蛋白组成的病毒样颗粒（VLPs）因其出色的安全性和免疫原性，作为一种新型疫苗受到了广泛关注。然而，与大多数疫苗抗原类似，VLPs的热稳定性不足，这不仅限制了疫苗的广泛应用，还增加了疫苗失活的风险。本研究旨在通过采用真空冷冻干燥技术提高A型口蹄疫病毒（FMDV）来源的VLPs的稳定性和保质期。通过单因素和组合筛选确定了最佳冻干保护剂配方。随后，通过小鼠模型评估冻干疫苗的免疫原性与FMDV VLPs含量之间的相关性。通过在4℃、25℃和37℃下储存不同时间段，进一步评估冻干前后FMDV VLPs的稳定性。结果表明，由7.5%海藻糖、0.1%吐温80、50 mmol/L甘氨酸、1%谷氨酸钠和3%聚乙烯吡咯烷酮（PVP）组成的1号冻干保护剂配方在真空冷冻干燥过程中有效保留了FMDV VLPs的含量。小鼠免疫试验表明，冻干FMDV VLPs诱导的特异性抗体、免疫球蛋白G1（IgG1）、白细胞介素-4（IL-4）和中和抗体水平与非冻干FMDV VLPs诱导的水平相当。热处理结果表明，冻干FMDV VLPs在4℃、25℃和37℃下储存的时间明显长于非冻干FMDV VLPs。总之，所选的冻干保护剂配方有效提高了FMDV VLPs疫苗的稳定性。本研究为提高新型亚单位疫苗的稳定性提供了有价值的见解。
