College Veterinary Medicine, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety, Yangzhou University, Yangzhou 225009, China.
Int J Mol Sci. 2024 Aug 15;25(16):8906. doi: 10.3390/ijms25168906.
The African swine fever virus (ASFV) is a large enveloped DNA virus that causes a highly pathogenic hemorrhagic disease in both domestic pigs and wild boars. The ASFV genome contains a double-stranded DNA encoding more than 150 proteins. The ASFV possesses only one protease, pS273R, which is important for virion assembly and host immune evasion. Therefore, the specific monoclonal antibody (mAb) against pS273R is useful for ASFV research. Here, we generated two specific anti-pS273R mAbs named 2F3 and 3C2, both of which were successfully applied for ELISA, Western blotting, and immunofluorescence assays. Further, we showed that both 2F3 and 3C2 mAbs recognize a new epitope of N terminal 1-25 amino acids of pS273R protein, which is highly conserved across different ASFV strains including all genotype I and II strains. Based on the recognized epitope, an indirect ELISA was established and was effective in detecting antibodies during ASFV infection. To conclude, the specific pS273R mAbs and corresponding epitope identified will strongly promote ASFV serological diagnosis and vaccine research.
非洲猪瘟病毒（ASFV）是一种大型包膜 DNA 病毒，可导致家猪和野猪发生高致病性出血性疾病。ASFV 基因组包含编码超过 150 种蛋白的双链 DNA。ASFV 只拥有一种蛋白酶 pS273R，它对病毒粒子组装和宿主免疫逃避很重要。因此，针对 pS273R 的特异性单克隆抗体（mAb）可用于 ASFV 研究。在此，我们生成了两种针对 pS273R 的特异性 mAb，分别命名为 2F3 和 3C2，它们均成功应用于 ELISA、Western blot 和免疫荧光检测。此外，我们表明 2F3 和 3C2 mAb 识别 pS273R 蛋白 N 端 1-25 个氨基酸的新表位，该表位在不同 ASFV 毒株中高度保守，包括所有基因型 I 和 II 毒株。基于识别的表位，建立了间接 ELISA，可有效检测 ASFV 感染期间的抗体。总之，所鉴定的特异性 pS273R mAb 和相应表位将极大地促进 ASFV 的血清学诊断和疫苗研究。
