文献类型：期刊文献

英文题名：Highly efficient and economical baculovirus expression system for preparing human papillomavirus type16 virus-like particle

作者：Zheng, J; Ma, J; Yang, XF; Liu, HL; Cheng, HW; Si, LS; Wang, YL

第一作者：Zheng, J

通讯作者：Wang, YL[1]

机构：[1]Xi An Jiao Tong Univ, Key Lab Biomed Informat Engn, Minist Educ, Inst Canc Res,Sch Life Sci & Technol, Xian 710061, Peoples R China

通讯机构：[1]corresponding author), Xi An Jiao Tong Univ, Key Lab Biomed Informat Engn, Minist Educ, Inst Canc Res,Sch Life Sci & Technol, Xian 710061, Peoples R China.

年份：2004

卷号：36

期号：8

起止页码：548-552

外文期刊名：ACTA BIOCHIMICA ET BIOPHYSICA SINICA

收录：;Scopus(收录号：2-s2.0-7244261639);WOS:【SCI-EXPANDED(收录号:WOS:000223682000005)】；

语种：英文

外文关键词：human papillomavirus; HPV16L1; protein purification; virus-like particle (VLP)

摘要：To improve the existing human papillomavirus type 16 (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 6xHis tag, and harvested 72 h postinfection (p.i.) at 27 degreesC. The ProBond(TM) purification system was used for protein purification. The molecular weight of expressed HPV16L1 protein was 5 8 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 2x10(7) 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 6xHis tag could self-assemble intovirions in vitro. Hopefully, the present expression and purification system is promising to be convenient, timesaving and economical for preparation of HPV16 VLP vaccine.