Recombinant dengue virus type 2 envelope/hepatitis B surface antigen hybrid protein expressed in Pichia pastoris can function as a bivalent immunogen
Recombinant dengue virus type 2 envelope/hepatitis B surface antigen hybrid protein expressed in Pichia pastoris can function as a bivalent immunogen
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DOI：
10.1016/s0168-1656(02)00181-5
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发表时间：
2002-10-23
影响因子：
4.1
通讯作者：
Khanna, N
中科院分区：
文献类型：
--
作者：
Bisht, H;Chugh, DA;Khanna, N
在甲醇营养型酵母Pichia pastoris中表达了编码前395个氨基酸残基的截短型登革病毒2型包膜蛋白（Den 2 E）以及与全长226个氨基酸的乙型肝炎表面抗原（B HBsAg）蛋白的读框内融合的Den 2 E。两种重组蛋白均显示出形成高分子量聚集体的能力的证据。纯化蛋白的电子显微镜分析表明，虽然Den 2 E显示无定形形态，Den 2 E-HBsAg作为结构良好的病毒样颗粒（VLP）存在。使用免疫金电子显微镜，这些VLP被证明含有Den 2 E-HBsAg杂合蛋白的两种组分。血清学分析表明，杂交VLP可以在体内作为双价免疫原发挥作用，其可以引起针对杂交蛋白的两种组分的免疫应答，如通过ELISA、免疫沉淀和免疫荧光数据所证明的。(C)2002 Elsevier Science B. V.保留所有权利。
A truncated version of the dengue virus type 2 envelope protein (Den2E) encoding the first 395 amino acid (aa) residues, and Den2E fused in-frame with the full-length 226-aa hepatitis B surface antigen (Den2E-HBsAg) protein were expressed in the methylotrophic yeast, Pichia pastoris. Both the recombinant proteins showed evidence of the capacity to form high molecular weight aggregates. Electron microscopic analysis of the purified proteins showed that while Den2E displayed an amorphous morphology, Den2E-HBsAg existed as well-structured virus-like particles (VLPs). Using immuno-gold electron microscopy, these VLPs were demonstrated to contain both components of the Den2E-HBsAg hybrid protein. Seroanalysis showed that the hybrid VLPs could function in vivo as bivalent immunogens, which could elicit immune responses directed against both components of the hybrid protein, as evidenced by ELISA, immunoprecipitation and immunofluorescence data. (C) 2002 Elsevier Science B.V. All rights reserved.