An anchored monopodial DNA walker triggered by proximity hybridization for amplified amperometric biosensing of nucleic acid and protein

This work designed an anchored monopodial DNA walker to amplify amperometric biosensing signal for sensitive detection of nucleic acid and protein. The biosensing surface was constructed by self-assembling hairpin DNA1 (H1) and small amount of P1-W (probe DNA1 hybridized with walking DNA) on a gold electrode. In the presence of target molecule, the walker could be triggered by the surface proximity hybridization product of P1, target and P2 to induce the cyclic hybridization of H1 with ferrocene modified hairpin DNA2 (H2-Fc), which took electroactive Fc to the electrode surface for amplified amperometric detection of the target. By linking P1 and P2 with dual specific DNA strands, aptamers or antibodies to recognize the target for proximity hybridization of P1 and P2, the walker amplified amperometric strategy could be used for highly sensitive biosensing of different targets. Using DNA and thrombin as the target models, the proposed biosensing methods achieved the linear range from 0.2 pM to 2 nM with a detection limit of 0.11 pM and 1.0 pM to 10 nM with a detection limit of 0.61 pM, respectively. The specific recognition process endowed the strategy with high selectivity and potential applications.