CAJ | 학술논문

目的筛选针对狂犬病毒人源基因工程抗体.方法采集狂犬疫苗免疫人群外周血并分离淋巴细胞,构建抗狂犬病毒Fab噬菌体抗体库,用纯化狂犬病毒CTN株富集筛选,ELISA初步鉴定阳性克隆,测序确定抗体轻重链型别,构建IgG全抗表达载体,转染293T细胞并纯化IgG抗体,采用竞争ELISA、Western Blot和IFA鉴定抗体功能.结果筛选出1株抗狂犬病毒的人源Fab抗体并表达纯化IgG全抗体,IFA结果显示抗体特异针对狂犬病毒,竞争性ELISA,Western Blot结果显示新筛抗体针对磷蛋白.结论首次通过Fab噬菌体抗体库技术获得针对狂犬病毒磷蛋白人源抗体,为磷蛋白功能机制研究和狂犬病的诊断提供了新的抗体.
목적 사선침대광견병독인원기인공정항체.방법 채집광견역묘면역인군외주혈병분리림파세포,구건항광견병독Fab서균체항체고,용순화광견병독CTN주부집사선,ELISA초보감정양성극륭,측서학정항체경중련형별,구건IgG전항표체재체,전염293T세포병순화IgG항체,채용경쟁ELISA、Western Blot화IFA감정항체공능.결과 사선출1주항광견병독적인원Fab항체병표체순화IgG전항체,IFA결과현시항체특이침대광견병독,경쟁성ELISA,Western Blot결과현시신사항체침대린단백.결론 수차통과Fab서균체항체고기술획득침대광견병독린단백인원항체,위린단백공능궤제연구화광견병적진단제공료신적항체.
Objective To obtain recombinant human anti-RVP genetically engineered antibodies from rabiator-derived antibody phage library.Metbods A combinatorial human Fab library against rabies virus was constructed with antibody genes harvested from peripheral blood of rabiators with high titer of antibody.The library was panned with phage display and selected by purified CTN-1 virus particles.After that the selected antibody was converted to full human IgG antibody with recombinant mammal cell antibodyexpression system.Results Unique 1 human Fab antibodies specific for RVP were obtained and conformed by competitive ELISA,Western Blotting and IFA.IFA,competitive ELISA and Western blotting showed the panned antibody 1A2 bound specifically to RVP.Conclusion Human antibody against RVP was firstly obtained through phage antibody library,and could be used for RVP functional mechanism research and rabies diagnosis.