West Nile virus is a widespread mosquito-borne human and animal pathogenic virus of increasing importance. The E protein of the viral envelope is critical for attachment and entry into the host cell and has been the target for vaccine design and small molecule inhibitors. Furthermore, the detection of anti-E IgM and IgG antibodies is widely used in serology to diagnose these infections. Here we describe a strategy for the production of recombinant antibodies against the E protein of West Nile virus for research and immunodiagnostic purposes. Initially the fast and easy protocol previously developed for the similar Tick-borne encephalitis virus has been adapted to West Nile virus E antigen production and purification. A human naïve scFv phage library has been selected on the produced antigen, identifying a panel of highly specific anti-E protein antibodies. Once produced as scFv-Fc recombinant proteins, the selected antibodies have been characterized by mapping their binding sites and by defining their affinity for the target. The impact on neutralizing virus attachment and entry has been also evaluated. The obtained results demonstrate the potential of the produced reagents for research and diagnostic applications.

中文翻译：

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针对西尼罗河病毒E蛋白的高度特异性重组抗体的选择和表征。

西尼罗河病毒是一种广泛传播的蚊媒人类和动物致病性病毒，日益重要。病毒包膜的E蛋白对于附着和进入宿主细胞至关重要，并且已成为疫苗设计和小分子抑制剂的目标。此外，抗E IgM和IgG抗体的检测在血清学中被广泛用于诊断这些感染。在这里，我们描述了一种针对西尼罗河病毒E蛋白的重组抗体的生产策略，用于研究和免疫诊断。最初，先前为类似的Tick传播性脑炎病毒开发的快速简便的方案已经适应于西尼罗河病毒E抗原的生产和纯化。已在产生的抗原上选择了人类纯净的scFv噬菌体文库，鉴定一组高度特异性的抗E蛋白抗体。一旦产生为scFv-Fc重组蛋白，就可以通过绘制其结合位点并定义其对靶标的亲和力来表征所选抗体。还评估了对中和病毒附着和进入的影响。获得的结果证明了所生产的试剂在研究和诊断应用中的潜力。