聚丙烯酰胺凝胶电泳分离恶性疟原虫pfs25膜蛋白的免疫原性研究

Immunogenicity investigation for plasmodium falciparum transmission-blocking vaccine antigen pfs25 isolated by polyacrylamide Gel Electrophoresis

目的:研究聚丙烯酰胺凝胶电泳法纯化的pfs25蛋白的免疫原性变化。方法:利用聚丙烯酰胺凝胶电泳法(SDS-PAGE)分离了毕赤酵母表达的恶性疟原虫pfs25膜蛋白,按照常规程序将分离的pfs25膜蛋白免疫小鼠,以验证其免疫原性。通过荧光光谱分析、酶联免疫吸附实验,分析了聚丙烯酰胺凝胶电泳过程中pfs25蛋白质免疫原性的变化情况。结果:SDS-PAGE方法成功分离了pfs25蛋白,免疫小鼠后不能产生特异性抗体;与pfs25标准品相比,纯化的pfs25蛋白的荧光光谱发生了改变,但是ELISA结果均为阳性。结论:聚丙烯酰胺凝胶电泳法纯化的pfs25蛋白保留了其免疫反应性,丧失了免疫原性。

Objective：pfs25 protein as the candidate antigen of Plasmodium falciparum transmission blocking vaccine was produced by recombinant gene expression in Pichia and purified by polyacrylamide gel electrophoresis（SDS-PAGE）.Methods： Isolation pfs25 protein has been investigated by fluorescence spectra and enzyme-linked immunosorbent assay.Results： According to a conventional procedure,the purified recombinant pfs25 protein immunized mice to verify its immunogenicity.Conclusion： The reasons which pfs25 protein immunogenicity was lost during SDS-PAGE purification were discussed.