中国兽医科学2024，Vol.54Issue(10)：1351-1358,8.DOI:10.16656/j.issn.1673-4696.2024.0152
猪圆环病毒2d型病毒样颗粒疫苗的制备及发酵工艺研究
Study on preparation and fermentation technology of porcine circovirus 2d virus-like particles vaccine
摘要
Abstract
To develop a porcine circovirus type 2d virus-like particles vaccine and evaluate its immunization effect,the PCV2d Cap protein were expressed by prokaryotic expression system and confirmed through SDS-PAGE and Western-blot analysis.When the prime conditions of fermentation process were screened,the His-SUMO tag of expressed PCV2d Cap was removed to make its self-assembly into VLPs in vitro.The morphological characterization of the VLPs was checked by transmission electron microscopy and nanoparticle-size analyzer.The antibody and cytokine levels in sera were detected at 28 d after mice immunized with the VLPs.The results showed that the highest level of protein expression was achieved at 0.5 mmol/L IPTG and a 12％N source feed rate,and the VLPs were found to have a particle diameter of approximately 20-25 nm.The titers of specific antibody and neutralizing antibody were more than 1∶512 and 1∶32 respectively,and the levels of IL-4 and IFN-γ cytokines were 80 μg/mL,which were significantly higher than those in the PBS group.In conclusion,this study successfully con-structed and obtained PCV2d VLPs using the prokaryotic expression system,which induced good cellular and humoral immunity and laid the groundwork for research on the porcine circovirus 2d subtype and porcine circovirus multivalent vaccine.关键词
猪圆环病毒2d型/病毒样颗粒/大肠杆菌表达系统/发酵工艺Key words
porcine circovirus 2d/virus-like particles/E.coli expression system/fermentation technology分类
农业科技引用本文复制引用
王运航,白满元,敬晓棋,吕熙,段梦迪,魏甜,李毅,张雅雅,陈相杨,郭慧琛,孙世琪..猪圆环病毒2d型病毒样颗粒疫苗的制备及发酵工艺研究[J].中国兽医科学,2024,54(10):1351-1358,8.基金项目
2023兰州市人才创业创新项目(2023-RC-3) （2023-RC-3）
甘肃省在站博士后专项(23JRRA554) （23JRRA554）
甘肃省科技重大专项(23ZD-NA007) （23ZD-NA007）
国家自然基金青年基金项目(32301127) （32301127）
陕西省科技厅重点研发项目(2023-YBNY-143) （2023-YBNY-143）
陕西省教育厅2023年度重点科学研究计划项目-协同创新中心项目(23JY090) （23JY090）
榆林学院研究生创新基金项目(2022YLYCX12) （2022YLYCX12）
国家重点研发计划项目(2022YFD1800603,2021YFD1800300) （2022YFD1800603,2021YFD1800300）
