Batten Carrie A, Clarke Ian N, Kempster Sarah L, Oliver Stefan L, Bridger Janice C, Lambden Paul R
Molecular Microbiology Group, University Medical School, Southampton General Hospital, Southampton SO16 6YD, UK.
Virology. 2006;356(1-2):179-87. doi: 10.1016/j.virol.2006.07.034. Epub 2006 Aug 24.
The Southampton norovirus (SV) capsid protein was expressed as VLPs by recombinant baculoviruses in insect cells and was used to immunize mice for the production of monoclonal antibodies (mAbs). One mAb, CM54, showed broad cross-reactivity to genogroup I (GI) noroviruses, but was not reactive to GII capsid proteins. Interestingly mAb CM54 reacted to a bovine norovirus capsid protein. Immunoblot analysis indicated the binding site for CM54 was located in the shell domain between amino acid residues 102-225 of the SV capsid protein. The epitope was mapped to high resolution using a peptide array and was located to the sequence LEDVRN at amino acid residues 162-167. Alignment of norovirus capsid protein sequences confirmed the epitope sequence was common to particular groups of human and bovine noroviruses. Modeling of the epitope onto the recombinant NV capsid protein revealed it was located to the inner surface of the shell domain.
南安普敦诺如病毒(SV)衣壳蛋白通过重组杆状病毒在昆虫细胞中表达为病毒样颗粒(VLPs),并用于免疫小鼠以产生单克隆抗体(mAb)。一种单克隆抗体CM54对基因I群(GI)诺如病毒表现出广泛的交叉反应性,但对GII衣壳蛋白无反应。有趣的是,单克隆抗体CM54与一种牛诺如病毒衣壳蛋白发生反应。免疫印迹分析表明,CM54的结合位点位于SV衣壳蛋白氨基酸残基102 - 225之间的壳结构域。使用肽阵列将表位定位到高分辨率,其位于氨基酸残基162 - 167处的序列LEDVRN。诺如病毒衣壳蛋白序列比对证实,该表位序列在特定组的人类和牛诺如病毒中是常见的。将该表位在重组NV衣壳蛋白上进行建模显示,它位于壳结构域的内表面。