Regardless of the promising use of nanoparticles (NPs) in biomedical applications, several toxic effects have increased the concerns about the safety of these nanomaterials. Although the pathways for NPs toxicity are diverse and dependent upon many parameters such as the nature of the nanoparticle and the biochemical environment, numerous studies have provided evidence that direct contact between NPs and biomolecules or cell membranes leads to cell inactivation or damage and may be a primary mechanism for cytotoxicity. In such a context, this work focused on developing a fast and accurate method to characterize the interaction between NPs, proteins and lipidic membranes by surface plasmon resonance imaging (SPRi) technique. The interaction of gold NPs with mimetic membranes was evaluated by monitoring the variation of reflectivity after several consecutive gold NPs injections on the lipidic membranes prepared on the SPRi biochip. The interaction on the membranes with varied lipidic composition was compared regarding the total surface concentration density of gold NPs adsorbed on them. Then, the interaction of gold and silver NPs with blood proteins was analyzed regarding their kinetic profile of the association/dissociation and dissociation constants (koff). The surface concentration density on the membrane composed of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine and cholesterol (POPC/cholesterol) was 2.5 times higher than the value found after the injections of gold NPs on POPC only or with dimethyldioctadecylammonium (POPC/DDAB). Regarding the proteins, gold NPs showed preferential binding to fibrinogen resulting in a value of the variation of reflectivity that was 8 times higher than the value found for the other proteins. Differently, silver NPs showed similar interaction on all the tested proteins but with a variation of reflectivity on immunoglobulin G (IgG) 2 times higher than the value found for the other tested proteins.

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.