A simple, sensitive SERS-based immunoassay realized in aqueous solution is demonstrated with a sandwich immune protocol. In such an immunoassay, antibodies-immobilized silica nanoparticles served as the immune substrate while 4MBA-labeled immuno-Au nanoparticles are used as the immune sensors. According to the TEM images, it is clear that the immune gold nanoparticles are embedded onto the surfaces of the silica nanoparticles specifically after the immunoreaction. As a result, the aggregations of gold nanoparticles have been formed with SERS-active "hot spots" on the dimers or multimers. The SERS results confirm that the method proposed in this paper is an effective way for SERS-based aqueous immunoassay and that the detection limit is as low as 0.1 ng/mL.