Gopinath Shreya, Hosamani Madhusudan, Basagoudanavar Suresh, Sreenivasa B P, Joseph Biljo V, Patil Sharanagouda S
CHRIST UNIVERSITY, Bengaluru, Karnataka, India.
ICAR-Indian Veterinary Research Institute, Bengaluru, Karnataka, India.
Vet Immunol Immunopathol. 2025 Aug;286:110965. doi: 10.1016/j.vetimm.2025.110965. Epub 2025 Jun 13.
Classical swine fever (CSF), caused by the classical swine fever virus (CSFV; genus Pestivirus), is a highly transmissible disease in pigs that leads to severe economic losses for pig farmers worldwide. Early and accurate diagnosis of CSFV infections is crucial for effective containment of the disease, especially in endemic areas. In this study, the full-length viral protein E was successfully expressed in E. coli. The recombinant protein was purified using Ni-NTA affinity chromatography, and its antigenicity confirmed by western blot and indirect ELISA (iELISA) using CSFV-specific antiserum. The purified E protein was then used as an immunogen to generate murine hybridomas producing monoclonal antibodies (mAbs). A panel of 15 mAbs targeting this protein was characterized using iELISA and western blotting, for their ability to react with native antigens in cultures infected with CSFV. These mAbs have the potential to be used in the development of diagnostic tools, particularly for the detection and titration of virus-infected cultures through endpoint dilution assays.
经典猪瘟（CSF）由经典猪瘟病毒（CSFV；瘟病毒属）引起，是猪的一种高度传染性疾病，给全球养猪户造成严重经济损失。对CSFV感染进行早期准确诊断对于有效控制该疾病至关重要，尤其是在流行地区。在本研究中，病毒全长蛋白E在大肠杆菌中成功表达。重组蛋白通过镍-亚氨基二乙酸（Ni-NTA）亲和层析进行纯化，并用CSFV特异性抗血清通过蛋白质免疫印迹法和间接酶联免疫吸附测定（iELISA）确认其抗原性。然后将纯化的E蛋白用作免疫原，以产生分泌单克隆抗体（mAb）的小鼠杂交瘤。使用iELISA和蛋白质免疫印迹法对一组针对该蛋白的15种mAb进行了表征，以评估它们与感染CSFV的培养物中的天然抗原发生反应的能力。这些mAb有潜力用于开发诊断工具，特别是通过终点稀释法检测和滴定病毒感染的培养物。
