Development of an amperometric immunosensor for the quantification of staphylococcus aureus using self-assembled monolayer-modiried electrodes as immobilization platforms

Amperometric immunosensors for the detection and quantification of S. aureus using MPA self-assembled monolayer modified electrodes for the immobilization of the immunoreagents are reported. Two different immunosensor configurations were compared. A competitive mode, in which protein A-bearing S. aureus cells and antiRbIgG labeled with horseradish peroxidase (HRP) compete for the binding sites of RbIgG immobilized onto the 3mercaptopropionic acid (MPA) modified electrode, was evaluated. Moreover, a sandwich configuration in which S. aureus cells were immobilized onto the MPA SAM, and RbIgG and antiRbIgG labeled with HRP were further linked to the electrode surface, was also tested. In both cases, TTF was used as the redox mediator of the HRP reaction with H2O2, and it was co-immobilized onto the MPA-modified gold electrode. After optimization of the working variables for both configurations, the analytical performance of the amperometric measurements carried out at 0.00 V (vs. Ag/ AgCl) showed that the competitive immunosensor exhibited a lower limit of detection (1.6 x 105 S aureus cells mL(-1)), as well as a better repeatability and reproducibility of the measurements.