The current studies on molecular mechanism of bivalves interacting with bacteria are mainly on mRNA and recombinant protein levels. These works provide little information on natural proteins, which limit further understandings. In this study, we conducted a pioneer work to promote researches on the anti-Vibrio immune response of Zhikong Scallop Chlamys farreri through proteomic techniques. Firstly a reference map was constructed for the hepatopancreas of C. farreri. Totally 65 protein spots were included in the reference map, while 46 of them were identified. Gene ontology analysis revealed high activities of metabolism and immunity in hepatopancreas. Furthermore, hepatopancreas of C. farreri injected with Vibrio harveyi at 24 h post-injection (hpi) were used for comparative proteomic analysis. Totally 27 differentially expressed proteins spots after challenge were screened; and 15 were successfully identified. These proteins include some immune-related proteins, metabolism enzymes, and new molecules which were not paid attentions in previous immunity studies in C. farreri. The results indicated that molecular chaperons and the antioxidant system are key elements in the anti-Vibrio immune response of hepatopancreas of C. farreri. The identification of new molecules provides indications for further studies. The results of this work provide a new insight into the anti-Vibrio immune response of marine bivalves.