Dendritic structure DNA for specific metal ion biosensor based on catalytic hairpin assembly and a sensitive...

In this work, a sensitive electrochemical biosensing to Pb²⁺ was proposed based on the high specificity of DNAzymes to Pb²⁺. The response signal was efficiently amplified by the catalytic hairpin assembly induced by strand replacement reaction and the formation of dendritic structure DNA (DSDNA) by layer-by-layer assembly. Firstly, in the presence of Pb²⁺, the substrate strand (S1) of the Pb²⁺-specific DNAzymes was specifically cleaved by Pb²⁺. Secondly, one of the two fragments (rS1) introduced into the electrode surface was hybridized with a hairpin DNA (H1) and further replaced by another hairpin DNA (H2) by the hybridization reaction of H1 with H2. The released rS1 then induced the next hybridization with H1. After repeated cycles, the catalytic recycling assembly of H2 with H1 was completed. Thirdly, two bioconjugates of Pt@Pd nanocages (Pt@PdNCs) labeled with DNA S3/S4 and electroactive toluidine blue (Tb) (Tb-S3-Pt@PdNCs and Tb-S4-Pt@PdNCs) were captured onto the resultant electrode surface through the hybridization of S3 and H2, S3 and S4, resulting in the formation of DSDNA triggered by layer-by-layer assembly. This formed DSDNA greatly facilitated the immobilization of manganese(III) meso-tetrakis (4-N-methylpyridiniumyl)-porphyrin (MnTMPyP) as mimicking enzyme. Under the synergistic catalysis of Pt@PdNCs and MnTMPyP to H₂O₂ reduction, the effective signal amplification of the developed Pb²⁺ biosensor was achieved. As a result, the sensitive detection of the proposed electrochemical strategy for Pb²⁺ was greatly improved in the range of 0.1pM-200nM with a detection limit of 0.033pM.