为表达抗鸡传染性法氏囊病病毒(IbdV)抗体fAb并检测其中和活性,本研究将抗IbdV抗体的轻链(l)和重链片段(fd)基因分别克隆于P ET-27b(+)载体中,并转化于大肠杆菌rOSETTA(dE3)进行诱导表达。将l和fd片段包涵体蛋白变性后等量混合于复性液中,制备fAb并对其进行活性鉴定。结果显示l和fd蛋白相对分子质量大小分别为25 ku和28 ku。WESTErn blOT和ElISA检测结果表明,获得的抗体fAb大小约为50 ku,并且与VP2蛋白和不同病毒株均具有特异性结合能力。体外中和试验结果表明,获得的IbdV抗体fAb具有中和活性,可以有效阻断IbdV(b87株)对鸡胚成纤维细胞(df1)的感染。本实验获得的IbdV抗体fAb有望成为治疗Ibd的候选生物制剂,为研制治疗Ibd抗体制剂奠定了基础。To express the neutralizing Fab antibody against infectious bursal disease virus(IBDV),the gene of light chain(L)or heavy chain fragment(Fd) against IBDV was cloned into the prokaryotic expression plasmid,respectively,and then the recombinant L or Fd was expressed in E.coli Rosetta(DE3),respectively,and purified through sole denaturation and co-renaturation of inclusion body.Western blot results showed that the Fab was approximately 50 ku.ELISA results showed that the Fab exhibited binding ability and specify to VP2 for different IBDV strains.The results of neutralization test in vitro showed that the Fab exhibited neutralizing activity to IBDV-B87 strainin chicken embryo fibroblast(DF1) cells.The Fab antibody prepared in this study is expected to become a candidate drug for treatment of IBD,which laid the foundation for the treatment of IBD.黑龙江省应用技术研究与开发计划项目(GC13C104
