A novel strategy based on antigen-antibody interaction was developed by means of backfilling transferrin on antibody functionalized gold nanoparticles surfaces in the present study. Nano gold particles were immobilized with cysteamine layer by self-assembly, whose surfaces were chemically coupled with anti-transferrin antibodies by using EDC, forming the anti-transferrin-Au immuno-probes. The particles and the nano-probes were characterized by the integrated tools of resonance Rayleigh scattering (RRS), UV-Vis absorption spectra, transmission electron microscopy (TEM) and laser light scattering. The result showed that the nano-probe with the ability of specific recognition of transferrin had good immune activity. The RRS peak at 470 nm was amplified obviously by using this signal amplification, and the antigen recognition was monitored via the enhancement of 470 nm RRS intensity when this binding event occurred. The result showed that the amplification strategy led to a dramatic improvement of the detection sensitivity of transferrin, and the detection of transferrin featured a linear range of 0.85 to 33.9×10-10 mol·L-1 with the detection limit of 8.5×10-11 mol·L-1.