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DNA Enzyme-Decorated DNA Nanoladders as Enhancer for Peptide Cleavage-Based Electrochemical Biosensor
- Kou, Bei-Bei, Zhang, Li, Xie, Hua, Wang, Ding, Yuan, Ya-Li, Chai, Ya-Qin, Yuan, Ruo
- ACS Applied Materials & Interfaces 2016 v.8 no.35 pp. 22869-22874
- biosensors, catalytic activity, detection limit, electrodes, hybridization chain reaction, hydrogen peroxide, iodine, manganese, nanocarriers, nanoparticles, oxidation, platinum, porphyrins, proteinases, quantitative analysis, single-stranded DNA
- Herein, we developed a label-free electrochemical biosensor for sensitive detection of matrix metalloproteinase-7 (MMP-7) based on DNA enzyme-decorated DNA nanoladders as enhancer. A peptide and single-stranded DNA S₁-modified platinum nanoparticles (P₁-PtNPs-S₁), which served as recognition nanoprobes, were first immobilized on electrode. When target MMP-7 specifically recognized and cleaved the peptide, the PtNPs-S₁ bioconjugates were successfully released from electrode. The remaining S₁ on electrode then hybridized with ssDNA1 (I₁) and ssDNA2 (I₂), which could synchronously trigger two hybridization chain reactions (HCRs), resulting in the in situ formation of DNA nanoladders. The desired DNA nanoladders not only were employed as ideal nanocarriers for enzyme loading, but also maintained its catalytic activity. With the help of hydrogen peroxide (H₂O₂), manganese porphyrin (MnPP) with peroxidase-like activity accelerated the 4-chloro-1-naphthol (4-CN) oxidation with generation of insoluble precipitation on electrode, causing a very low differential pulse voltammetry (DPV) signal for quantitative determination of MMP-7. Under optimal conditions, the developed biosensor exhibited a wide linear ranging from 0.2 pg/mL to 20 ng/mL, and the detection limit was 0.05 pg/mL. This work successfully realized the combination of DNA signal amplification technique with artificial mimetic enzyme-catalyzed precipitation reaction in peptide cleavage-based protein detection, offering a promising avenue for the detection of other proteases.