目的利用重组博尔纳病病毒核蛋白进行动物免疫,制备多克隆抗体并对其进行鉴定。方法将重组载体pET14b-p40转化至感受态大肠埃希菌I,PTG诱导融合蛋白的表达,His-tag亲和层析纯化重组核蛋白并作为抗原免疫新西兰大白兔,收集免疫后血清,制备和纯化多克隆抗体,ELISA测定抗体效价,并进行Western-blot鉴定。结果成功制备出核蛋白多克隆抗体,ELISA检测效价高达1︰256000;该抗体与原核和真核系统中表达的核蛋白均能发生特异性反应。结论成功制备了效价和特异性良好的抗重组核蛋白多克隆抗体,为博尔纳病病毒血清免疫学检测方法的建立奠定了基础。 Objective To immunize animals with recombinant Borna disease virus nucleoprotein to prepare polyclonal antibodies and identify them. Methods Recombinant vector pET14b-p40 was transformed into competent Escherichia coli I, the expression of fusion protein was induced by PTG. His-tag affinity chromatography was used to purify recombinant nucleoprotein and immunized New Zealand White rabbits as antigen. The polyclonal antibody was purified, the antibody titers were determined by ELISA and identified by Western-blot. Results The polyclonal antibody of nucleoprotein was successfully prepared. The ELISA titer was up to 1︰256000. The antibody reacted specifically with the expressed nucleoprotein in prokaryotic and eukaryotic cells. Conclusion The polyclonal antibody against recombinant nucleoprotein with good titer and specificity was successfully prepared and laid the foundation for the establishment of immunological detection of Borna disease virus.