Wash-free photoelectrochemical DNA detection based on photoredox catalysis combined with electroreduction and light blocking by magnetic microparticles

To obtain a sensitive, wash-free photoelectrochemical biosensor based on electron mediation between an electrode and a photoredox catalyst (PC) label, unavoidable O2-related reactions should have no effect or be beneficial, and the rate of electron mediation should depend on the distance between the PC label and electrode. A wash-free photoelectrochemical biosensor that (i) combines photoredox catalysis of a PC label with electrochemical reduction of an electron mediator, and (ii) uses a light-blocking multilayer of magnetic microparticles was developed. O2 participates as an electron acceptor in photoredox catalysis; thus, increasing rather than decreasing the electrochemical signal. Upon photoirradiation from the opposite side of a transparent indium tin oxide (ITO) electrode in contact with the solution, the light intensity in the solution is sharply decreased by the light-blocking multilayer, which increases the contribution of affinity-bound PC labels on the ITO electrode to the electrochemical signal compared to that of unbound PC labels in solution. Utilizing eosin Y (EY2−) and Fe(CN)64− as the PC and electron mediator (i.e., electron donor), respectively, enabled rapid redox cycling based on photoredox catalysis combined with electroreduction. The cathodic charge is mainly related to electron transfer from Fe(CN)64− to excited EY2− (Type I photosensitization), rather than energy transfer from excited EY2− to O2, which generates 1O2 (Type II photosensitization). The developed detection scheme was applied to wash-free detection of a model target DNA. Detection limits of ∼200 pM were obtained in both phosphate-buffered saline and serum without washing. The developed scheme enables simple photoelectrochemical detection. [Display omitted] •Wash-free photoelectrochemical biosensor using a photoredox catalyst label.•Utilizing eosin Y and Fe(CN)64− as the label and electron mediator, respectively.•Dissolved O2 as an electron acceptor in photoredox catalysis.•Use of multilayer of magnetic microparticles for distance-dependent light intensity.•Sharply distance-dependent electron mediation enables wash-free DNA detection.