Sensors based on the surface plasmon resonance (SPR) technique are useful devices to detect and monitor interactions between biomolecules in real-time. SPR is a label-free method that monitors the variation of reflectivity of a biochip composed of a metal-coated glass prism and can be applied in several areas, such as biotechnology, food safety and clinical diagnosis. In the last years, several researchers have proven the efficiency of metallic nanoparticles (NPs) in the enhancement of SPR signal. This feature allowed the detection of biomolecules at very low concentration. Aiming to further enhance SPR signal towards the detection of proteins at low concentration and by a simple procedure, the present work compared the performance of gold and platinum bimetallic NPs (AuPtNPs) with that of monometallic gold NPs (AuNPs) in the enhancement of SPR signal. In order to evaluate the NPs, protein peanut agglutin (PNA) was used as target analyte and anti-PNA antibody was used as sensing molecule. Firstly NPs were functionalized with anti-PNA antibody and incubated with a solution containing PNA. Then, the NPs bound to PNA were injected into the SPR equipment containing a biochip previously modified with anti-PNA antibody. The results demonstrated that the AuPtNPs provided a 91-fold increase compared to the direct detection of free PNA in solution. In comparison with AuNPs, the signal generated by AuPtNPs was about 4 times higher. This encouraging result indicated that the application of bimetallic NPs may be a better strategy to further enhance sensitivity of SPR biosensors and could drive the development of new strategies that are not only simple, but also able to detect proteins at low concentrations, which is of great importance, especially in clinical diagnostics.