Xi Xiangfeng, Mo Xiaobing, Xiao Yan, Yin Bo, Lv Chaochao, Wang Yuzhou, Sun Zhe, Yang Qingyuan, Yao Yali, Xuan Yajie, Li Xiangdong, Yuan Y Adam, Tian Kegong
National Research Center for Veterinary Medicine, Road Cuiwei, High-Tech District, Luoyang 471003, PR China.
National University of Singapore (Suzhou) Research Institute, 377 Lin Quan Street, Suzhou Industrial Park, Jiangsu 215123, PR China; Department of Biological Sciences and Centre for Bioimaging Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore, Singapore.
J Biotechnol. 2016 Apr 10;223:8-12. doi: 10.1016/j.jbiotec.2016.02.025. Epub 2016 Feb 22.
We report the strategies leading to the large-production of soluble non-tag full-length porcine circovirus type 2 (PCV2) Cap protein in Escherichia coli. Under neutral pH condition, the purified recombinant Cap protein derived from E. coli expression self-assembles into homogenous round virus-like particle at the similar size of that of the intact PCV2 virus, which is further characterized by Cryo-EM single particle structure determined at 4.5Å. The engineered PCV2 rCap VLP was tested as a subunit vaccine for the protective efficacy against PCV2 challenge on 3-week old piglets. Similar to commercial available PCV2 vaccine, the Cap VLP-immunized piglets developed specific antibody-mediated response and were protected from the virulent SH PCV2 strain challenge. Hence, the production of E. coli based PCV2Cap-VLP could be applied as a cost-friendly and effective subunit vaccine to control PCV2 spreading in developing countries.
我们报道了在大肠杆菌中大量生产可溶性无标签全长猪圆环病毒2型（PCV2）Cap蛋白的策略。在中性pH条件下，从大肠杆菌表达中纯化得到的重组Cap蛋白自组装成大小与完整PCV2病毒相似的均匀圆形病毒样颗粒，通过冷冻电镜单颗粒结构在4.5埃分辨率下进一步表征。将工程化的PCV2 rCap VLP作为亚单位疫苗，测试其对3周龄仔猪PCV2攻击的保护效果。与市售PCV2疫苗相似，用Cap VLP免疫的仔猪产生了特异性抗体介导的反应，并免受强毒SH PCV2毒株的攻击。因此，基于大肠杆菌生产的PCV2Cap-VLP可作为一种经济友好且有效的亚单位疫苗，用于控制发展中国家PCV2的传播。
