http://www.abbs.info E-mail: [email protected]
ISSN
1672-9145
Acta Biochim Biophys Sin
2004, 36(8): 548–552
CN 31-1940/Q
Highly Efficient and Economical Baculovirus Expression System for Preparing Human Papillomavirus Type16 Virus-like Particle
Jin ZHENG, Jun MA, Xiao-Feng YANG, Hong-Li LIU, Hong-Wei CHENG, Lu-Sheng SI, and Yi-Li WANG*
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
Abstract 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 2107 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.
Key words human papillomavirus; HPV16L1;
protein purification; virus-like particle (VLP)
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Received: March 31, 2004 Accepted: June 23, 2004
This study was supported by a grant from the Ministry of Science and Technology of China (No. 2001AA215221)
*Corresponding author: Tel, 86-29-82655499; Fax, 86-29-82655499; E-mail, [email protected]