非洲猪瘟,由非洲猪瘟病毒(ASFV)诱导,由于其在家猪和野猪中的高杀伤力,对全球养猪业构成了重大威胁。尽管疾病严重,缺乏针对ASFV的有效疫苗和药物。p72蛋白,占总病毒颗粒质量的31%至33%,作为ASFV的主要衣壳蛋白。它是开发ASF亚单位疫苗和血清学诊断方法的关键抗原。在这次调查中,通过用大肠杆菌表达的截短的C末端p72蛋白进行小鼠免疫产生27种单克隆抗体(mAb)。其中,六个单克隆抗体表现出与p72三聚体的结合,它们各自识别的表位被鉴定为542VTAHGINLIDKF553、568GNAIKTP574和584FALKPREEY592。所有三个表位都位于p72的C末端果冻卷桶的功能单元的间隔序列内。值得注意的是,两个表位,568GNAIKTP574和584FALKPREEY592在p72三聚体内部,虽然表位542VTAHGINLIDKF553暴露在三聚体的表面,并且在所有ASFV基因型中始终保守。这些发现增强了我们对p72蛋白的抗原功能和结构的理解,促进p72在ASFV诊断技术开发中的应用。
African swine fever, which is induced by the African swine fever virus (ASFV), poses a significant threat to the global pig industry due to its high lethality in domestic pigs and wild boars. Despite the severity of the disease, there is a lack of effective vaccines and drugs against the ASFV. The p72 protein, constituting 31 to 33% of the total virus particle mass, serves as the primary capsid protein of ASFV. It is a crucial antigen for the development of ASF subunit vaccines and serological diagnostic methods. In this investigation, 27 monoclonal antibodies (mAbs) were generated through mouse immunization with the truncated C-terminal p72 protein expressed by Escherichia coli. Among these, six mAbs exhibited binding to the p72 trimer, with their respective recognized epitopes identified as 542VTAHGINLIDKF553, 568GNAIKTP574, and 584FALKPREEY592. All three epitopes were situated within the interval sequences of functional units of the C-terminal jelly-roll barrel of p72. Notably, two epitopes, 568GNAIKTP574 and 584FALKPREEY592, were internal to the p72 trimer, while the epitope 542VTAHGINLIDKF553 was exposed on the surface of the trimer and consistently conserved across all ASFV genotypes. These findings enhance our comprehension of the antigenic function and structure of the p72 protein, facilitating the utilization of p72 in the development of diagnostic techniques for ASFV.