Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70 (HSP70)-peptide complexes (PCs) from human melanoma cell lines A375, A875, M21, M14, WM-35, and SK-HEL-1. We named the purified product as M-HSP70-PCs and determined its immunological activities. Autologous HSP70-PCs purified from primary tumor cells of melanoma patients (9 cases) were used as controls. These two tumor antigenic complexes were loaded into dendritic cells (DCs) and used to stimulate an antitumor response against tumor cells in the corresponding patients. Results Mature DCs pulsed with M-HSP70-PCs stimulated autologous T cells to secrete the same levels of type I cytokines as the autologous HSP70-PCs. Moreover, DCs pulsed with M-HSP70-PCs endued CIK cells with an equal ability as autologous HSP70-PCs to kill melanoma cells in the patients. Conclusion M-HSP70-PCs may be used as an efficient and generalized tumor antigen in the treatment of DC-based malignant melanoma.