Highly efficient and economical baculovirus expression system for preparing human papillomavirus type16 virus-like particle.
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 6xHis tag, and harvested 72 h postinfection (p.i.) at 27 degrees. The ProBond(TM) 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 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 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.
The 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
J
Zheng
Jun
J
Ma
Xiao-Feng
XF
Yang
Hong-Li
HL
Liu
Hong-Wei
HW
Cheng
Lu-Sheng
LS
Si
Yi-Li
YL
Wang
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D002460:Cell Line; D015870:Gene Expression; D005814:Genes, Viral; D005822:Genetic Vectors; D006801:Humans; D007181:Inclusion Bodies, Viral / Q000648:ultrastructure; D008854:Microscopy, Electron; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D018411:Spodoptera; D014162:Transfection
D036022:Capsid Proteins; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; C063317:L1 protein, Human papillomavirus type 16