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对毕赤酵母 (Pichiapastoris)表达的重组乙肝表面抗原SS1的纯化以及纯化蛋白质的理化性质及免疫原性进行了进一步的研究。采用单抗亲和层析的方法 ,一步纯化即可得到纯度为 95 %的纯化蛋白质。ELISA和Western印迹鉴定表明 ,纯化的SS1融合蛋白同时具有良好的S和PreS1抗原性。CsCl密度梯度离心和电镜检测的结果则表明 ,这一重组抗原可以形成与天然HBV亚病毒颗粒类似的颗粒结构。在BALB/c小鼠中进行的免疫实验表明 ,纯化的SS1融合蛋白可以同时激起很强的抗HBs和抗PreS1的抗体反应。使用CpG免疫佐剂 ,可以促使免疫小鼠产生针对HBs和PreS1的Th1类免疫反应
Purification of the recombinant hepatitis B surface antigen SS1 expressed by Pichia pastoris and the physicochemical properties and immunogenicity of the purified protein were further studied. Using monoclonal antibody affinity chromatography method, one-step purification to obtain a purity of 95% of the purified protein. ELISA and Western blotting showed that the purified SS1 fusion protein had both good S and PreS1 antigenicity. CsCl density gradient centrifugation and electron microscopy results show that this recombinant antigen can form a particle structure similar to the natural HBV subviral particles. Immunization experiments in BALB / c mice showed that the purified SS1 fusion protein simultaneously elicited strong anti-HBs and anti-PreS1 antibody responses. Using CpG immunoadjuvant, the immune mice can be induced to produce Th1-like immune responses against HBs and PreS1