HIV-1病毒样颗粒的纯化及其免疫原性

目的纯化HIV-1病毒样颗粒,并检测其免疫原性。方法稳定表达HIV-1结构蛋白Gag和Env的重组293细胞经大规模培养后,收集培养上清,经30%蔗糖垫两次纯化,免疫BALB/c小鼠,Western blot检测小鼠血清中抗体水平。结果重组293细胞培养上清的纯化产物经SDS-PAGE分析,可见目的蛋白的表达。Western blot检测显示,纯化蛋白具有良好的反应原性。免疫小鼠后能够诱导产生针对目的蛋白的抗体,其抗体水平呈剂量依赖关系。结论纯化的HIV-1病毒样颗粒能够诱导小鼠产生抗体,具有良好的免疫原性。

Purification and Immunogenicity of HIV-1-like Particles

Objective To purify HIV-1-like particles and determine their immunogenicity.Methods Culture the recombi-nant 293 cells for stable expression of HIV-1 structural protein Gag and Env in a large scale,collect the culture supernatant and puri-fy by ultracentrifugation using 30% sucrose.Immunize BALB / c mice with the purified protein and determine the serum neutralizing antibody by Western blot.Results SDS-PAGE showed that target protein was expressed in the culture supernatant of recombinant 293 cells.The purified expressed protein showed good reactogenicity as proved by Western blot and induced specific antibody in mice in a dose-dependent mode.Conclusion HIV-1-like particles induced antibody in mice and showed good immunogenicity.