The African Swine Fever Virus (ASFV) poses a significant threat to the global pig farming industry. The major icosahedral capsid protein p72 plays a key role in the assembly of virus particles and infection process. As one major antigen of ASFV, p72 has been widely utilized as a marker for diagnosing infection. In this study, five p72 specific monoclonal antibodies (mAbs) were generated through the immunization of mice followed by cell fusion. Among the five hybridomas, clones 1B7, 2F3, 5D3, and 5D4 recognized a novel epitope of