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首页> 美国卫生研究院文献>Clinical and Experimental Immunology >Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1
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Selection of single chain variable fragments (scFv) against the glycoprotein antigen of the rabies virus from a human synthetic scFv phage display library and their fusion with the Fc region of human IgG1

机译：从人合成的scFv噬菌体展示文库中选择针对狂犬病病毒糖蛋白抗原的单链可变片段（scFv）并将其与人IgG1的Fc区融合

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摘要

We have prepared human recombinant antibody molecules against the glycoprotein antigen of the rabies virus (GPRV) based on the single chain variable fragment (scFv) format. Anti-GPRV scFvs were selected from a human synthetic scFv phage display library with a repertoire of approximately 109 specificities. After three rounds of selection against the PV11 strain of the virus, 40% of the clones tested recognized the rabies antigen. Of the 20 positive clones that were sequenced, five distinct sequences were identified. These distinct scFvs were cloned into a mammalian expression vector carrying the human IgG1 Fc region. The specificity of the resulting scFv-Fc molecules for GPRV was established by ELISA, dot blot and western blot analyses and membrane immunofluorescence. Two of the scFv-Fc fusion proteins neutralized the PV11 strain in a standard in vivo neutralization assay where the virus was incubated with the scFv-Fc molecules before intracranial inoculation in mice. These anti-GPRV scFv-Fc molecules have the potential to be used as an alternative to the presently available HRIG, for use in post-exposure preventive treatment.
机译：我们已经基于单链可变片段（scFv）格式制备了针对狂犬病毒（GPRV）糖蛋白抗原的人重组抗体分子。从人合成的scFv噬菌体展示文库中选择具有约10 9 特异性的抗GPRV scFv。经过三轮针对病毒PV11株的选择后，测试的克隆中有40％识别出狂犬病抗原。在测序的20个阳性克隆中，鉴定出5个不同的序列。将这些不同的scFvs克隆到带有人IgG1 Fc区的哺乳动物表达载体中。通过ELISA，斑点印迹和蛋白质印迹分析以及膜免疫荧光确定了所得的scFv-Fc分子对GPRV的特异性。 scFv-Fc融合蛋白中的两种在标准体内中和测定中中和了PV11菌株，在该实验中，将病毒与scFv-Fc分子温育后再进行小鼠颅内接种。这些抗GPRV scFv-Fc分子有潜力用作目前可用的HRIG的替代品，用于暴露后预防治疗。

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