The aim of the study was to generate B cell epitope maps of the hexon L1 using monoclonal antibodies (mAbs) and bioinformatics tools. In this study, the hexon L1 (230 amino acids) was expressed and used to generate 10 clones of mAbs against the relative protein peptide. Furthermore, we defined the linear peptide epitopes recognized by these mAbs using a series of partially overlapping peptides derived from the FAdV-C hexon protein amino acid sequence to map mAbs reactivity. Finally, a common B cell epitope (31PLAPKESMFN40) for all species FAdVs and two FAdV-C-specific epitopes (79KISGVFPNP87and181DYDDYNIGTT190) were identified. These mAbs and their defined epitopes may support the development of the universal or species-specific differential diagnostic methods of FAdVs.