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为制备高效价、高特异性的裂谷热病毒（(Rift Valley fever virus， RVFV）非结构蛋白NSs多克隆抗体，并评估其应用价值，本研究采用大肠杆菌原核表达系统表达并纯化了RVFV NSs重组蛋白。以该纯化蛋白作为免疫原免疫BALB/c小鼠，成功获得了抗NSs多克隆抗体。经间接ELISA检测，该抗体效价达1：512000以上；Western blot及间接免疫荧光实验结果显示，所获抗体能同时特异性识别原核和真核系统表达的NSs蛋白，表明其对变性及天然构象的NSs蛋白均具有良好的反应特异性。综上，本研究成功制备了高效价、高特异性的RVFV NSs小鼠多克隆抗体，为深入研究NSs蛋白的亚细胞定位、在病毒感染致病过程中的功能以及筛选抗病毒药物靶点提供了关键工具。
Abstract：Polyclonal antibodies are essential tools for studying viral protein function. However， a high-quality antibody against the nonstructural protein NSs of Rift Valley fever virus (RVFV)， a key virulence determinant， remains unavailable. In this study， we expressed and purified recombinant NSs protein of RVFV (strain ZH501) using an E. coli prokaryotic expression system. The purified protein was then used as an immunogen to immunize BALB/c mice， successfully generating a murine anti-NSs polyclonal antibody. Indirect ELISA demonstrated that the antibody titer exceeded 1:512，000. Furthermore， Western blot and indirect immunofluorescence assays demonstrated that the obtained antibody specifically recognized the NSs protein expressed in both prokaryotic and eukaryotic systems.， confirming its strong reactivity against both denatured and native conformations of NSs. In conclusion， we have generated a high-titer， highly specific polyclonal antibody against RVFV NSs protein. This antibody serves as a valuable tool for investigating NSs subcellular localization， elucidating its role in viral pathogenesis， and screening for antiviral drug targets.
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基本信息:
中图分类号:S852.65
引用信息:
[1]王丹,肖萌萌,贺怀悦,等.裂谷热病毒非结构蛋白NSs的原核表达、纯化及多克隆抗体制备[J].经济动物学报().
基金信息:
国家自然科学基金项目（32473121）
2026-03-20
2026-03-20
2026-03-20