Mercury or silver was electrodeposited on an Au surface to form an Hg-Au or Ag-Au film. Wavelength-modulation SPR biosensors based on this Hg/Ag-Au film were then used to determine human IgG and rabbit IgG. When direct immunoreactions were performed on the Au sensing surface, the range of concentrations of human IgG and rabbit IgG that could be determined were 2.00-40.00 microg/ml and 2.50-40.00 microg/ml, respectively. When Hg was electrodeposited onto the Au film for 1200 s, the range of concentrations of human IgG and rabbit IgG that could be determined were 0.50-40.00 microg/ml and 0.63-40.00 microg/ml, respectively. When Ag was electrodeposited onto the Au film for 1500 s, the range of concentrations of human IgG and rabbit IgG that could be determined were 0.25-20.00 and 0.42-20.00 microg/ml, respectively. The biosensor based on Ag-Au film was therefore found to be the most sensitive of the three types of biosensor tested, giving limits of determination that were up to eight times lower than those obtained with a biosensor based on Au film alone.