Department of Genetic Engineering, Graduate School of Biotechnology, Kyung Hee University, Yongin, 446-701, Korea.
Plant Cell Rep. 2011 Aug;30(8):1513-21. doi: 10.1007/s00299-011-1062-6. Epub 2011 Mar 27.
We describe the expression and immunogenicity of a recombinant chimeric protein (HAV VP1-Fc) consisting of human hepatitis A virus VP1 and an Fc antibody fragment using a replicating vector based on Beet curly top virus (BCTV) in Agrobacterium-infiltrated Nicotiana benthamiana leaves. Recombinant HAV VP1-Fc was expressed with a molecular mass of approximately 68 kDa. Recombinant HAV VP1-Fc, purified using Protein A Sepharose affinity chromatography, elicited production of specific IgG antibodies in the serum after intraperitoneal immunization. Following vaccination with recombinant HAV VP1-Fc protein, expressions of IFN-γ and IL-4 were increased in splenocytes at the time of sacrifice. Recombinant VP1-Fc from infiltrated tobacco plants can be used as an effective experimental immunogen for research into vaccine development.
我们使用基于甜菜曲顶病毒（BCTV）的复制载体在农杆菌浸润的黄花烟草叶片中描述了由人甲型肝炎病毒 VP1 和 Fc 抗体片段组成的重组嵌合蛋白（HAV VP1-Fc）的表达和免疫原性。重组 HAV VP1-Fc 的分子量约为 68 kDa。使用 Protein A Sepharose 亲和层析法纯化重组 HAV VP1-Fc，在腹腔免疫后血清中产生特异性 IgG 抗体。用重组 HAV VP1-Fc 蛋白免疫接种后，在牺牲时脾细胞中 IFN-γ 和 IL-4 的表达增加。浸润烟草植物的重组 VP1-Fc 可作为疫苗开发研究的有效实验免疫原。
