目的建立去除抗肝癌基因疫苗pVAX1-aLCGV内毒素的方法。方法采用Triton X-114液相分离法去除抗肝癌基因疫苗中的内毒素,鲎试剂法检测疫苗中的内毒素含量,家兔法复试,ELISA法检测内毒素入血前后血浆中细胞因子TNF-α和IL-8的含量及疫苗的免疫原性,并比较处理前后的基因疫苗转染SMMC-7721和HepG2细胞的转染效率。结果经Triton X-114处理后,基因疫苗的内毒素含量及热原检测均合格;内毒素入血后,血浆中TNF-α和IL-8的含量均明显升高;疫苗的免疫原性与处理前比较,差异无统计学意义;处理后疫苗质粒的细胞转染效率增高。结论 Triton X-114能有效去除基因疫苗pVAX1-aLCGV中的内毒素,为其临床应用奠定了基础。 Objective To establish a method for removing endotoxin of anti-HCC gene vaccine pVAX1-aLCGV. Methods Triton X-114 liquid phase separation was used to remove the endotoxin in the anti-hepatoma gene vaccine. The LPS assay was used to detect the endotoxin content in the vaccine. Rabbit retesting and ELISA were performed to detect the levels of cytokine TNF- α and IL-8 and the immunogenicity of the vaccine, and compared the transfection efficiency of SMMC-7721 and HepG2 cells transfected with the gene vaccine before and after treatment. Results After treatment with Triton X-114, the levels of endotoxin and pyrogen in the gene vaccines were both good. The content of TNF-α and IL-8 in the plasma was significantly increased after the endotoxin was infused into the blood. The immunogenicity of the vaccine Before treatment, the difference was not statistically significant; the efficiency of transfection of the vaccine plasmids after treatment increased. Conclusion Triton X-114 can effectively remove endotoxin in gene vaccine pVAX1-aLCGV, which lays the foundation for its clinical application.