Topology of bovine rotavirus (RF strain) VP6 epitopes by real-time biospecific interaction analysis.
Topology of bovine rotavirus (RF strain) VP6 epitopes by real-time biospecific interaction analysis.
复制标题
通过实时生物特异性相互作用分析牛轮状病毒（RF 株）VP6 表位的拓扑。
DOI：
10.1006/viro.1994.1505
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发表时间：
1994
期刊：
影响因子：
3.7
通讯作者：
J. Cohen
中科院分区：
文献类型：
--
作者：
G. Tosser;T. Delaunay;E. Kohli;J. Grosclaude;P. Pothier;J. Cohen
设计用于实时测量分子相互作用的自动化生物传感器系统（BIAcore）用于表征针对牛轮状病毒（RF株）的内衣壳蛋白（VP6）产生的单克隆抗体（Mab）。六个单克隆抗体，所有反应在Western印迹和间接免疫荧光测定，映射，使用纯化的重组VP6。这些单克隆抗体被划分为几组抗体。通过BIAcore方法研究了所选单克隆抗体与纯化病毒颗粒的相互作用。我们发现一些单克隆抗体不与单壳颗粒反应。相反，几种单克隆抗体与单壳颗粒反应，一种抗体与单壳和双壳颗粒反应。后一种单抗似乎通过外衣壳的孔与VP6相互作用，并且其与双壳颗粒的相互作用诱导了显著的去衣壳作用。这些结果允许更好地表征VP6的表位。在病毒颗粒中的表位的本地化进行了讨论，与pepscan的研究，确定与来自整个VP6分子的嵌套heptapeptides的单克隆抗体的反应性比较。
An automated biosensor system designed for measuring molecular interactions in real-time (BIAcore) was used to characterize monoclonal antibodies (Mabs) raised against the inner capsid protein (VP6) of the bovine rotavirus (RF strain). Six Mabs, all reactive in Western blot and in indirect immunofluorescence assays, were mapped, using purified recombinant VP6. These Mabs were delineated into several groups of antibodies. Interactions of selected monoclonal antibodies with purified viral particles were studied by the BIAcore methodology. We showed that some Mabs did not react with single-shelled particles. Conversely, several Mabs reacted with single-shelled particles and one antibody reacted with both single-shelled and double-shelled particles. The latter Mab seemed to interact with VP6 through the holes of the outer capsid and its interaction with the double-shelled particles induced a significant decapsidation. These results allowed a better characterization of the epitopes of VP6. The localization of the epitopes in the viral particle is discussed in comparison with a pepscan study that determined the reactivity of Mabs with nested heptapeptides derived from the whole VP6 molecule.