Raux H, Iseni F, Lafay F, Blondel D
Laboratoire de Génétique des Virus, CNRS, Gif Sur Yvette, France.
J Gen Virol. 1997 Jan;78 ( Pt 1):119-24. doi: 10.1099/0022-1317-78-1-119.
Thirty-six monoclonal antibodies (MAbs) specific for the rabies virus P phosphoprotein were obtained from mice immunized with recombinant P (PV strain) produced in E. coli. All MAbs reacted against the corresponding rabies virus protein by ELISA and by Western blot analysis and revealed the presence of cytoplasmic inclusions in rabies virus infected cells. The epitopes of seven MAbs were mapped by testing their reactivity with protein fragments expressed from deletion mutants in transfected cells. Western blotting, immunoprecipitation and immunofluorescence assays were performed. These MAbs recognized epitopes in different domains of the P protein: 60% were directed against a region lying between residues 83-172 suggesting a major antigenic determinant of the rabies virus P protein in this region. Most of the antigenic sites appeared to be composed of linear epitopes. These MAbs will be useful as tools to dissect structural and functional properties of the rabies virus P protein.
用在大肠杆菌中产生的重组P（PV株）免疫小鼠，获得了36种针对狂犬病病毒P磷蛋白的单克隆抗体（MAb）。通过酶联免疫吸附测定（ELISA）和蛋白质免疫印迹分析，所有单克隆抗体均与相应的狂犬病病毒蛋白发生反应，并揭示了狂犬病病毒感染细胞中存在细胞质内含物。通过测试七种单克隆抗体与转染细胞中缺失突变体表达的蛋白质片段的反应性，对其表位进行了定位。进行了蛋白质免疫印迹、免疫沉淀和免疫荧光测定。这些单克隆抗体识别P蛋白不同结构域中的表位：60%针对位于83-172位氨基酸之间的区域，表明该区域是狂犬病病毒P蛋白的主要抗原决定簇。大多数抗原位点似乎由线性表位组成。这些单克隆抗体将作为剖析狂犬病病毒P蛋白结构和功能特性的工具。
