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目的为了获取人源性HFRS基因工程抗体。方法直接从人体PBL构建人全套ScFv、VH噬菌体表面呈现文库,以panning方法筛至四级文库,以ELISA方法鉴定各克隆抗汉坦病毒活性。结果获得了14个ScFv阳性克隆和11个VH阳性克隆。其中ScFv最高阳性克隆A410值为0.44,VH最高阳性克隆A410值为0.50。对最高阳性克隆进行了测序分析,证明连接和克隆正确,获得了抗HTV人源性ScFv和VH抗体的基因片段。克隆再转化于HB2151菌株,成功地进行了分泌型抗体片段的表达。结论采用噬菌体抗体库技术直接从人体克隆人源性汉坦病毒噬菌体抗体可行,对HFRS的防治有一定的实际意义
Purpose To obtain human-derived HFRS engineered antibodies. Methods The whole set of human ScFv was constructed directly from human PBL. The library of VH phage was displayed. The library was screened by panning method and the anti-Hantavirus activity of each clone was identified by ELISA. As a result, 14 ScFv positive clones and 11 VH positive clones were obtained. The highest ScFv positive clone A410 value was 0.44 and the highest VH clone positive clone A410 value was 0.50. Sequencing of the highest positive clones was performed to verify that the ligation and cloning were correct, and gene fragments of human anti-HTV-derived ScFv and VH antibodies were obtained. Clones were transformed into HB2151 strain and the expression of secreted antibody fragments was successfully performed. Conclusion It is feasible to clone human phage anti-Hantavirus antibody directly from human using phage antibody library technology, which has some practical significance for the prevention and treatment of HFRS