The purpose of the study is to identify the novel approach of hsp110-heparanase vaccine in virtue of the chaperoning properties of hsp110, and to explore the antitumor potential. To test it, the recombinant hsp110 and heparanase proteins form into complex vaccine during heat shock, and the antitumor immunity is observed. We demonstrate that hsp110 and heparanase can form into complex vaccine, and the vaccine can enhance the specific IFN-gamma production and cytotoxic T cell response. Additionally, immunization of the vaccine can also significantly suppress the established tumor growth and prolong the life span of tumor-bearing mice. Thus, the use of the recombinant hsp110 and heparanase proteins to form into complex vaccine represents a novel and promising way for the therapeutical strategy of tumor.