毕赤酵母表达的重组乙肝表面抗原SS1的纯化及性质鉴定  

Purification and Characterization of Recombinant Hepatitis B Virus Surface Antigen SS1 Expressed in Pichia pastoris

作　　者：杨剑莹[1] 金菁[1] 孔玉英[1] 卫军[1] 张祖传[1] 李光地[1] 汪垣[1] 袁汉英[2] 李育阳[2] 

机构地区：[1]中国科学院上海生物化学研究所,上海200031 [2]复旦大学遗传学研究所,上海200433

出　　处：《生物化学与生物物理学报》2000年第5期503-508,共6页

基　　金：国家高技术"863"资助项目 !No .Z18 0 1 0 2 0 2&&

摘　　要：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 。The purification of recombinant hepatitis B surface antigen, SS1 pro tein, expressed in Pichia pastoris, and the investigation of its physiochemi cal characters and immunogenicity were described here. Employing McAb immunoaffinity chromatography, this protein was purified to purity of 95%. The results of ELIS A and Western blotting showed good antigenicity of this purified SS1 protein. Cs Cl gradient centrifugation and electron microscopy assay proved that this purifi ed protein could be assembled into particles similar to the HBV subviral particl es. Strong antibody responses against both the HBs and PreS1 epitopes were induc ed in BALB/c mice immunized with this purified protein. The simultaneous injecti on of a CpG adjuvant induced a Th1-like immune response against both the HBs an d PreS1 epitopes.

关 键 词：乙肝表面抗原 PRES1 毕赤属酵母 纯化 性质 

分 类 号：R392.11[医药卫生—免疫学]