In this study, we have demonstrated that hemoglobin can be coupled to acid-treated multiwall carbon nanotubes in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and assembled as hemoglobin-carbon nanotube (Hb-CNT) composites. Our observations of the electrochemical studies demonstrate that the electrochemical response of Hb-CNT assembled in the presence of EDC is much higher than that in the absence of EDC. It is evident that the direct electron transfer of hemoglobin could be effectively accelerated in the Hb-CNT assembly by using EDC on a glassy carbon electrode (GCE), and the relative electron transfer rate constant K(s) is found to be 1.02+/-0.05 s(-1). The results of our studies illustrate that the assembly of hemoglobin-multiwall carbon nanotubes using EDC could provide a novel strategy to effectively facilitate the direct electrochemistry of heme-containing proteins, which could be further utilized as a promising biosensor for some specific biological substrate and related biological process.