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

•A Pb2+ biosensor with high specificity was constructed by using specific DNAzymes.•A Pt@PdNCs and MnTMPyP cooperatively amplified electrochemical response signal.•The signal was amplified by dendritic structure DNA and catalytic hairpin assembly.

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 0.1 pM–200 nM with a detection limit of 0.033 pM.