Human monoclonal antibodies in single chain fragment variable format with potent neutralization activity against influenza virus H5N1
Human monoclonal antibodies in single chain fragment variable format with potent neutralization activity against influenza virus H5N1
复制标题
DOI：
10.1016/j.antiviral.2009.05.005
复制
发表时间：
2009-09-01
影响因子：
7.6
通讯作者：
Cianfriglia, Maurizio
中科院分区：
文献类型：
--
作者：
Ascione, Alessandro;Capecchi, Barbara;Cianfriglia, Maurizio
需要有效的诊断和治疗策略来控制和对抗高致病性禽流感病毒 (AIV) H5N1 亚型。为此，我们开发了针对H5N1禽流感病毒的单链片段可变（scFv）形式的人单克隆抗体（mAb），为针对人类H5N1病例的免疫治疗的开发获得新的见解。使用基于生物淘选的方法，从人类半合成 ETH-2 噬菌体抗体库中分离出大量针对 H5N1 病毒的 scFv。具有独特的可变重链 (VH) 和轻链 (VL) 基因序列的 H5N1 ELISA 阳性 scFv 在 H5N1 病毒株中表现出不同的生化特性和中和活性。特别地，scFv克隆AV.D1和AV.C4在基于假型的中和测定中对H5N1 A/Vietnam/1194/2004病毒感染发挥显着抑制作用。有趣的是。这两种scFv相对于A/百日咳/蒙古/244/2005和A/印度尼西亚/5/2005菌株表现出跨进化枝中和活性。这些研究证明了这样一个概念：通过使用灭活的 H5N1 病毒作为诱饵对完全人工抗体库进行生物淘选，可以轻松快速地分离出具有明确 H5N1 识别模式和体外中和活性的 scFv 形式的人单克隆抗体。 (C) 2009 Elsevier B.V. 保留所有权利。
Effective diagnostic and therapeutic strategies are needed to control and combat the highly pathogenic avian influenza virus (AIV) subtype H5N1. To this end, we developed human monoclonal antibodies (mAbs) in single chain fragment variable (scFv) format towards the H5N1 avian influenza virus to gain new insights for the development of immunotherapy against human cases of H5N1. Using a biopanning based approach a large array of scFvs against H5N1 virus were isolated from the human semi-synthetic ETH-2 phage antibody library. H5N1 ELISA-positive scFvs with unique variable heavy (VH) and light (VL) chain gene sequences showed different biochemical properties and neutralization activity across H5N1 viral strains. In particular, the scFv clones AV.D1 and AV.C4 exerted a significant inhibition of the H5N1 A/Vietnam/1194/2004 virus infection in a pseudotype-based neutralization assay. Interestingly. these two scFvs displayed a cross-clade neutralizing activity versus A/whooping swan/Mongolia/244/2005 and A/Indonesia/5/2005 strains. These studies provide proof of the concept that human mAbs in scFv format with well-defined H5N1 recognition patterns and in vitro neutralizing activity can be easily and rapidly isolated by biopanning selection of an entirely artificial antibody repertoire using inactivated H5N1 virus as a bait. (C) 2009 Elsevier B.V. All rights reserved.