Electrochemiluminescence: insights into the mechanisms and nanomaterial-based applications

Biomarkers are biological indicators of human health conditions. Their ultra-sensitive quantification is of paramount importance in clinical monitoring and early disease diagnosis. Biosensors are simple and easy-to-use analytical devices and, in their world, electrochemiluminescence (ECL) is one of the most promising analytical techniques that needs an ever-increasing sensitivity for improving its clinical effectiveness. Scope of this project was the investigation of the ECL generation mechanisms for enhancing the ECL intensity also through the identification of suitable nanostructures. The combination of nanotechnologies, microscopy and ECL has proved to be a very successful strategy to improve the analytical efficiency of ECL in one of its most promising bioanalytical approaches, the bead-based immunoassay. Nanosystems, such as [Ru(bpy)3]2+-dye-doped nanoparticles (DDSNPs) and Bodipy Carbon Nanodots, have been used to improve the sensitivity of ECL techniques thanks to their advantageous and tuneable properties, reaching a signal increase of 750% in DDSNPs-bead-based immunoassay system. In this thesis, an investigation of size and distance effects on the ECL mechanisms was carried out through the innovative combination of ECL microscopy and electrochemical mapping of radicals. It allowed the discovery of an unexpected and highly efficient mechanistic path for ECL generation at small distances from the electrode surface. It was exploited and enhanced through the addition of a branched amine DPIBA to the usual coreactant TPrA solution for enhancing the ECL efficiency until a maximum of 128%. Finally, a beads-based immunoassay and an immunosensor specific for cardiac Troponin I were built exploiting previous results and carbon nanotubes features. They created a conductive layer around beads enhancing the signal by 70% and activating an ECL mechanism unobserved before in such systems. In conclusion, the combination of ECL microscopy and nanotechnology and the deep understanding of the mechanisms responsible for the ECL emission led to a great enhancement in the signal.