Expression and Purification and Particle Self-assembly of Novel Recombinant Nanoparticles Based on HBc: A Model Substrate for Exploring HBc as Anti-tumor Vaccine Carrier
Hepatitis B virus core (HBc) nanoparticles represent useful tools for display foreign epitopes. But little strategy has been reported to date on anti-tumor applicability of the HBc nanoparticle as carrier. Here we report high expression and purification of HBc-derived nanoparticle and exploit its use as a potential tumor vaccine carrier. The gene encoding for target oligopeptides for epidermal growth factor receptor (EGFR) was internally inserted into the HBc carrier by genetic engineering. The aimed plasmid was transformed into E. coli strain Rosseta (DE3) and overexpressed by induction of 1 mM IPTG, designated HBc-1L. The nanoparticle was purified by ammonium sulfate fractionation of the soluble bacterial lysate followed by fraction of particulate and unparticulate forms by sucrose gradient, and then by Sepharose CL-4B gel filtration chromatography. The separation effectiveness was confirmed by transmission electron microscopy, revealing the presence of spherical particles. The immunogenicity of HBc-1L was evaluated the serum of immunized mice by indirect ELISA. The inhibitory effects of antiserum were tested in a variety of human cancer cells by MTT assay. Mouse HBc-1L antiserum can significantly inhibit the growth of cells. These results illustrated the potential application of HBc nanoparticles as the anti-tumor vaccine carrier.
hepatitis B virus core (HBc) nanoparticles epidermal growth factor receptor antitumor
Jing Kong Xiao-Zhao Deng Xu Gu Zheng-yu Diao Hui Zhong Juan-wen Yao
Department of Life Science and Chemical Engineering, Huaiyin Industrial College, Huaiyin, China, 223 Huadong Research Institute for Medicine and Biotechnics, Nanjing, China,210009 School of Life Science and Technology, China Pharmaceutical University
国际会议
The 5th International Forum on Post-genome Technologies(5IFPT)(第五届国际后基因组生命科学技术学术论坛)
苏州
英文
2007-09-10（万方平台首次上网日期，不代表论文的发表时间）