Artificial receptors for the electrochemical detection of bacterial flagellar filaments from Proteus mirabilis

In this paper for the first time we successfully detect bacterial flagellar filaments from Proteus mirabilis using molecularly imprinted artificial receptors. These receptors acted as a sensing layer of the biosensors, assembled by imprinting flagellar proteins onto a polymeric backbone of electropolymerized phenol. In short, flagellar filaments were absorbed onto a carbon support, phenol was electropolymerized around it through the carbon conductive matrix to create the protein molecular molds, and finally the flagellar proteins were removed by enzymatic and electrochemical action. Each removed flagellar protein gave rise to an imprinted site with eventual rebinding ability. Electrical impedance spectroscopy (EIS) and square wave voltammetry (SWV) were employed to measure the interaction of flagellar filaments with the sensing layer assembled on commercial screen-printed electrodes, providing low detection limits, high precision and selectivity toward the targeted protein. The detection limit was 0.7 ng/mL by EIS and 0.9 ng/mL by SWV. The artificial receptors were further assembled on home-made paper-printed electrodes, with the three-electrode system printed on a paper substrate, offering the possibility of detecting flagellar filaments at as low as 0.6 ng/mL with a disposable and cost-effective portable device. To the best of our knowledge this is the first sensing device where molecularly imprinted artificial receptors are tailored on home-made electrode based on paper substrates with three electrodes assembled together, which is a suitable approach for the fabrication of easy and cost-effective tailored electrodes.