人源抗狂犬病毒G蛋白单链抗体的生物学鉴定及其小鼠体内中和活性测定

目的对从噬菌体抗体库中筛选的人源抗狂犬病毒单链抗体A12进行生物学活性鉴定及小鼠体内中和活性检测。方法用免疫荧光法检测其结合感染狂犬病毒CVS的鼠脑组织的能力,用小鼠中和实验测定A12表达产物的体内中和活性。结果免疫荧光试验显示A12表达产物与感染CVS的鼠脑细胞有强的荧光反应。小鼠中和试验结果表明,A12ScFv样品组小鼠有9只存活,而对照组小鼠全部死亡。A12在729倍稀释时能100%保护小鼠抵抗致死量狂犬病毒的脑内攻击。结论A12对狂犬病毒具有一定的中和活性,有可能被用于暴露后狂犬病的预防。

Biological identification of the humanized monoclonal single chain antibody against the glycoprotein of rabies virus and its in vivo neurtralization capacity in mouse model

The humanized monoclonal antibody ScFV-A12, selected from a human synthetic ScFV phage display library, was identified biologically, and its WTBX]in vivoWTBZ] neutralization capacity was determined in mouse model. The binding activity of ScFv-A12 to mouse brain tissues infected with rabies virus CVS strain was determined by immunofluorescence test, and the neutralization capacity of A12 was measured WTBX]in vivoWTBZ] by a standard neutralization assay in mouse model. It was found that the strong reactivity of ScFV-A12 to the mouse brain cells infected with rabies virus CVS could be demonstrated by IFA. In the mouse neutralization test, each 20 mice were used in the experimental and the control group in which 9 mice survived in ScFv-A12 treated group, while the mice died in the control group. In addition, ScFv-A12 could be 100% protection of mice against lethal challenge with rabies virus CVS inoculated intracerebrally at 1∶729 dilution of ScFv-A12, suggesting its potential use in the post-exposure preventive treatment.