In vitro screening assays are useful techniques for the determination of receptor-mediated activities in environmental samples. In order to define whether environmental chemicals act as an agonist or antagonist to the human estrogen receptor (hER), we have constructed a biosensor based on ligand-inducible interactions between hER and relative proteins on a quartz crystal microbalance (QCM). The his-tagged proteins, which were expressed in E. coli by recombinant DNA technology, were immobilized on an Au-electrode with Ni(II)-mediated chemisorption using the histidine tag and thiol-modified iminodiacetic acid. The resonance-frequency change of the protein-modified electrode was caused by association or dissociation with the hER relative proteins on the surface in the presence of estrogen. These results suggest that this sensor is applicable as a large-scale screening tool for estrogenic compounds.