Synthesis of nanoparticles with desired size/morphology has enormous importance, especially in the compelling field of nanotechnology. In this case, a novel kind of raspberry-type hybrid nanoparticles was prepared by hybridization of chitosan (CS) with thioethyl pendant ligand (TPL) modified Au nanoparticles (Au-TPL@CS NPs). Such method was based on ionic gelation using sodium tripolyphosphate as a counterion. The blood compatibility of Au-TPL@CS NPs was characterized by coagulation tests, plasma recalcification time, hemolysis assay, morphological changes of red blood cells (RBCs) and complement activation in vitro. The results showed that Au-TPL@CS NPs exhibited good blood compatibility. The possible underlying mechanism was also present. Finally, the direct electron transfer reactivity of the Hemoglobin/Au-TPL@CS NPs/multi-walled carbon nanotubes/glassy carbon electrode was investigated with cyclic voltammetry measurements. The biosensor exhibited a good electrocatalytic activity to the reduction of H2O2. Such new type of Au-TPL@CS NPs provides a promising platform of biological system for early illness detection and treatment in future.