毕氏酵母表达的EV71 VP1蛋白免疫原性研究

The immunogenicity of VP1 of enterovirus 71(EV71) expressed in yeast Pichia pastoris

目的研究毕氏酵母表达的肠道病毒71(EV71)VP1的免疫原性,为研究EV71的亚单位疫苗提供实验依据。方法用生物信息学分析EV71的VP1序列,优化在酵母表达的密码子。表达质粒p PICZa A-VP1电转毕氏酵母,并在甲醇诱导下表达VP1,VP1用Ni-NTA亲合层析得到纯化并用Western blot分析鉴定。用小鼠模型评估其免疫原性和疫苗效果。结果重组的VP1能有效诱导BALB/c小鼠产生抗-VP1抗体,免疫后的血清中抗VP1抗体能中和EV71病毒。对新生小鼠的被动保护进一步证实了VP1接种的小鼠抗血清的预防效应。结论重组VP1蛋白保留了其免疫原性,是一种候选的EV71疫苗。

Objective To study the immunogenicity of VP1 expressed in yeast Pichia pastoris, and provide experimental evidence of EV 71 subunit vaccine. Methods VP1 sequences were analyzed using bioinformatics and codes were optimized to express in yeast Pichia pastoris. The expression plasmid p PICZa A-VP1 was transformed and expressed in yeast Pichia pastoris induced by methanol. VP1 in the supernatant was purified using Ni-NTA affinity chromatography and characterized by western blot analysis. Immunogenicity and vaccine efficacy of there combinant VP1 were assessed in mouse models. Results The recombinant VP1 could efficiently induce anti-VP1 antibodies production in BALB / c mice,which were able to neutralize EV71 viruses. Passive protection of neonatal mice further confirmed the prophylactic efficacy of the antisera from VP1 vaccinated mice. Conclusion Recombinant VP1 protein retained good immunogenicity and was a potent EV71 vaccine candidate.