Highly Efficient and Economical Baculovirus Expression System for Preparing Human Papillomavirus Type16 Virus-like Particle

作     者：Jin ZHENG, Jun MA, Xiao-Feng YANG, Hong-Li LIU, Hong-Wei CHENG, Lu-Sheng SI, and Yi-Li WANGThe Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute of Cancer Research,School of Life Science and Technology, Xi’an Jiaotong University, Xi’an 710061, China Jin ZHENG, Jun MA, Xiao-Feng YANG, Hong-Li LIU, Hong-Wei CHENG, Lu-Sheng SI, and Yi-Li WANGThe Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute of Cancer Research,School of Life Science and Technology, Xi’an Jiaotong University, Xi’an 710061, China

作者机构：The Key Laboratory of Biomedical Information Engineering of Ministry of Education Institute of Cancer Research School of Life Science and TechnologyXi'an Jiaotong University Xi'an 710061 China 

出 版 物：《Acta Biochimica et Biophysica Sinica》 (生物化学与生物物理学报(英文版))

年 卷 期：2004年第36卷第8期

页      面：548-552页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：This study was supported by a grant from the Ministry of Science and[(T)64.3(echnology of China \(No. 2001AA215221\))] 

主　　题：human papillomavirus HPV16L1 protein purification virus-like particle (VLP) 

摘      要：To improve the existing human papillomavirus type16 (HPV16) virus-like particle (VLP) preparation, a highly efficient, economical and timesaving system was established. Sf-9 cells were infected with recombinant baculovirus containing the target gene encoding HPV16L1 protein with 6×His tag, and harvested 72 h postinfection (p.i.) at 27 °C. The ProBondTM purification system was used for protein purification. The molecular weight of expressed HPV16L1 protein was 58 kD as revealed by SDS-PAGE, and confirmed by Western blot. The purity of denatured and native HPVL1 proteins that were prepared were 91.9% and 71.5%, respectively, which corresponded to a yield of 2.26 mg denatured protein and 1.84 mg native protein per 2×107 cells. The proteins were further analyzed by mouse erythrocyte hemagglutination assay and hemagglutination inhibition assay, and there effects on VLP formation were also visualized by transmission electron microscopy. Results showed that the native protein purified was biologically active as natural HPVL1 protein, inducing the murine erythrocyte agglutination and VLP formation. In addition, the purified recombinant HPV16L1 native protein with 6×His tag could self-assemble into virions in vitro. Hopefully, the present expression and purification system is promising to be convenient, timesaving and economical for preparation of HPV16 VLP vaccine.