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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 of enterovirus 71 (EV71) expressed by Pichia pastoris and provide experimental evidence for the study of subunit vaccine of EV71. Methods The VP1 sequence of EV71 was analyzed by bioinformatics to optimize codons for expression in yeast. The plasmid p PICZa A-VP1 was transformed into Pichia pastoris and expressed VP1 under methanol induction. VP1 was purified by Ni-NTA affinity chromatography and identified by Western blot analysis. The mouse model was used to evaluate its immunogenicity and vaccine efficacy. Results Recombinant VP1 effectively induced anti-VP1 antibody in BALB / c mice, and the anti-VP1 antibody in the immunized serum could neutralize EV71 virus. Passive protection of neonatal mice further confirmed the prophylactic effect of VP1 vaccinated mouse antisera. Conclusion The recombinant VP1 protein retains its immunogenicity and is a candidate EV71 vaccine.