A Dynamic DNA Machine via Free Walker Movement on Lipid Bilayer for Ultrasensitive Electrochemiluminescent Bioassay

Herein, an ultrasensitive electrochemiluminescent (ECL) strategy was proposed based on a highly efficient dynamic DNA machine based on microRNA triggered free movement on the lipid bilayer interface. Typically, the lipid bilayer is constructed on the electrode surface modified with nafion@ECL luminophore and gold nanoparticles to immobilize the DNA walker labeled with cholesterol and hairpin nucleotides labeled with cholesterol and ferrocene (Fc), based on the cholesterol-lipid interaction. On this state, Fc was close to the ECL luminophore, performing a quenched ECL emission. In the presence of target microRNA 21, it could trigger the entropy beacon-based DNA amplification to convert microRNA to massive special DNA sequences, which could further hybridize with the blocking DNA on DNA walker to reactivate the DNA walker and thus trigger the DNA walker-based amplification to make Fc to be far from the ECL luminophore, performing a recovered ECL emission related with the concentration of microRNA 21. Compared with the conventional DNA walker immobilized on the interface via chemical bonds or physical adsorption, a higher reaction efficiency could be achieved due to the free movements of DNA walker and its substrates on the interface. As expected, satisfactory performances for the detection of microRNA 21 were achieved with a detection limit of 0.4 fM and quantitative estimation in cells. Furthermore, this dynamic DNA machine-based ECL strategy could be readily expanded for the detection of other biomarkers for clinical diagnosis.