Detection of circulating Asian H5N1 viruses by a newly established monoclonal antibody
Detection of circulating Asian H5N1 viruses by a newly established monoclonal antibody
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DOI：
10.1016/j.bbrc.2008.11.022
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发表时间：
2009-01-09
影响因子：
3.1
通讯作者：
Nakaya, Takaaki
中科院分区：
文献类型：
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作者：
Du, Anariwa;Daidoji, Tomo;Nakaya, Takaaki
产生了针对最近出现的亚洲H5 N1病毒（A/crow/京都/53/2004）的单克隆抗体（MAb）。从五种抗血凝素（HA）单克隆抗体中，四种抗体（3C 11、4C 12、3 H12和3 H4）在体外广泛识别并中和H5亚型，包括H5 N1。相比之下，4G 6单克隆抗体特异性地与H5 N1-HA反应，而不与来自先前流行的H5 N2-或H5 N3-HA反应。4G 6单克隆抗体可用于免疫荧光分析，但不能用于免疫印迹，表明该抗体识别H5 N1-HA蛋白的构象表位。一个密集的表位作图分析表明，4G 6单克隆抗体识别Asp 59，这是目前流行的H5 N1谱系中高度保守的。此外，基于4G 6的抗原捕获酶联免疫吸附测定检测到H5 N1，甚至是来自病毒分离前感染的鸡肺的进化枝2.2（A/chicken/Egypt/CL-61/2007）的H5 N1。总而言之，这些结果表明，已建立的单克隆抗体，尤其是4G 6，可用于快速、特异性检测亚洲H5 N1病毒。(C)2008年爱思唯尔公司All rights reserved.
Monoclonal antibodies (MAbs) against the recently emerged Asian H5N1 virus (A/crow/Kyoto/53/2004) were generated. From five anti-hemagglutinin (HA) MAbs, four antibodies (3C11, 4C12, 3H12, and 3H4) broadly in vitro recognized and neutralized H5 subtypes, including H5N1. By contrast, the 4G6 MAb specifically reacted with H5N1-HA and not with H5N2- or H5N3-HAs from previous epidemics. The 4G6 MAb was useful for immunofluorescence assays but not for immunoblotting, suggesting that this antibdy recognizes as conformational epitope of the H5N1-HA protein. An intensive epitope-mapping analysis demonstrated that the 4G6 MAb recognizes Asp59, which is highly conserved among currently circulating H5N1 lineages. Further, a 4G6-based antigen capture enzyme-linked immunosorbent assay detected H5N1 even that derived from clade 2.2 (A/chicken/Egypt/CL-61/2007) from infected chicken lung before virus isolation. Taken together, these results suggest that the established MAbs, especially 4G6, are useful for rapid and specific detection of Asian H5N1 viruses. (C) 2008 Elsevier Inc. All rights reserved.