Mimotope mapping enables the characterization of allergen epitopes for the development of diagnostic and therapeutic approaches. In the present study, a phage display peptide library was used for mimotope mapping based on the binding of antibodies against the recombinant group 5 allergen from the house dust mite Dermatophagoides farinae (Der f 5), an arthropod that causes indoor allergies worldwide. When three monoclonal anti-Der f 5 antibodies were used for biopanning, seven mimotopes were identified. Their common subsequence was '---[-A][-T] W[-S] H[HSFW][LM][PSKR] [TLV][AST] -[DP][-L] -'. When analyzed in combination with predicted discontinous epitopes, amino acids P2, K3, K4, H5, F11, F13, L14, R72, T77, L79, R84, T39, F40, P44, T45 and K46 were identified as key residues in conformational epitopes of Der f 5. Therefore, the seven mimotopes or modification of the key amino acids may facilitate the development of blocking antibodies or epitope-specific immunotherapies for mite allergy.