抗博尔纳病病毒核蛋白多克隆抗体的制备和鉴定

Preparation and identification of the polyclonal antibody against Borna disease virus nucleoprotein

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 prepare and identify the polyclonal antibody against Borna disease virus（BDV） nucleoprotein using recombinant BDV nucleoprotein. Method The recombinant vector PET14b-p40 was transformed into Escherichia coli BL21.The expression of recombinant nucleoprotein was induced by IPTG,and purified with His affinity chromatography.The purified target protein was used as antigen to immunize New Zealand rabbits to prepare the polyclonal antibody against BDV nucleoprotein,which was then characterized by indirect ELISA and Western blotting. Result The polyclonal antibody showed high affinity and obvious specificity and its titer was above 1︰256000.It had reacted specifically with the BDV nucleoprotein expressed both in prokaryotic and eukaryotic cells respectively. Conclusion This polyclonal antibody was prepared successfully,which may lay a foundation for further studies on BDV serum immunology detection methods.