Directionally In Situ Self-Assembled Iridium(III)-Polyimine Complex-Encapsulated Metal-Organic Framework Two-Dimensional Nanosheet Electrode To Boost Electrochemiluminescence Sensing.

Manufacturing electrochemiluminescence (ECL) electrodes to detect analytes with high performance in the aqueous phase for water-insoluble metal complexes is a great challenge. Here, a directional self-assembling avenue for in situ fabricating iridium(III)-polyimine complex-encapsulated metal-organic framework (MOF) two-dimensional electrode Hf-MOF/Ir2PD/APS/ITO is developed. The electrode displayed bright red ECL emission with high stability in the aqueous phase and specific adsorption toward ssDNA against dsDNA and mNs. That is to say, a "high-performance and multifunctional ECL electrode" is presented and explored for sensitive detection of acetamiprid (Ace) with a limit of detection of 0.0025 nM, where Ace-aptamer recognition-switched Exonuclease III-mediated digestion to make large numbers of Fc-labeled ssDNA transform into Fc-mNs. Furthermore, the proposed method was triumphantly employed to monitor the change in the residual concentration of Ace in pakchoi. This work breaks through the bottleneck of metal complex-based ECL emission in organic solvents and provides a novel strategy to develop high-performance ECL sensors.

Aptamer Recognition-Driven Homogeneous Electrochemical Strategy for Simultaneous Analysis of Multiple Pesticides without Interference of Color and Fluorescence.

Credible and simultaneous determination of multiple pesticides is highly desirable for guaranteeing food safety. However, the current methods are limited to significant interference of color and fluorescence or electrode's modification and mainly focus on the analysis of a single pesticide. Herein, we proposed a novel aptamer-based homogeneous electrochemical system for highly sensitive and simultaneous analysis of multiple pesticides based on target pesticide-switched exonuclease III (Exo III)-assisted signal amplification. The recognition of hairpin probes by target pesticides impels the production of pesticide-DNA complexes, which hybridize with electroactive dye-labeled DNA to form double-stranded DNA, subsequently initiating an Exo III-assisted digestion reaction to generate abundant electroactive dye-tagged mononucleotides. In comparison with pesticide deficiency, two higher differential pulse voltammetry (DPV) currents are measured, which rely on the amount of target pesticides. Therefore, simultaneous analysis of two pesticides is realized with limits of detection of 0.0048 and 0.0089 nM, respectively, comparable or superior to those of known methods that focused on a single pesticide. Moreover, the proposed system is successfully employed to simultaneously evaluate the residual level of acetamiprid and profenofos in Brassica chinensis and thus will find more useful applications for pesticide-related food safety.

N-heterocyclic Ir(III) complex targeting G-quadruplex structure to boost label-free and immobilization-free electrochemiluminescent sensing.

Development of simple, low-cost and highly sensitive electrochemiluminescence (ECL) sensors for various targets is urgent but remains challenging. It is because most of emitters were required to be dissolved in organic solvent or immobilized at electrode's surface to display ECL emission, and suffered from complicated tagging procedures and short emission wavelength. Herein, we synthesize an iridium (III) complex (Ir-ECL) and applied it as a ECL emitter for improved target sensing. ECL emission of Ir-ECL originated from the sensitization of N-heterocyclic ligands on Ir (III). Impressively, Ir-ECL exhibited ECL emission wavelength at 590 nm, and displayed a superior intercalation ability into G-quadruplex DNA against single-stranded DNA and double-stranded DNA (dsDNA). Using such properties, Ir-ECL was applied to enzyme-free, label-free, sensitive and homogeneous ECL analysis of pesticide acetamiprid (Ace) based on aptamer-target recognition-driven hybridization chain reaction (HCR). The recognition of H1 by Ace switched HCR of H2 and H3 to generate a long-chain dsDNA with abundant G-quadruplex DNAs, in which large numbers of Ir-ECL were locked, resulting in falling diffusion toward electrode, declining ECL signal and eventually improving Ace ECL sensing.

Aptamer recognition-promoted specific intercalation of iridium complexes in G-quadruplex DNA for label-free and enzyme-free phosphorescence analysis of kanamycin.

In consideration of relevance of antibiotic with food security, it is extremely desirable to propose sensitive and credible methods for antibiotic screening. Nevertheless, most of known approaches are developed based on fluorescence technique, which suffered from the interferences of background fluorescence and autoluminescence, and tedious labeling procedures, ascribing to the deficiency of high-performance and multifunctional dyes. Herein, we developed a novel iridium (III) complex (Ir-QAU)-based aptamer-promoted phosphorescence sensor for label-free, enzyme-free and highly sensitive detection of target antibiotic (kanamycin, Kan) based on target-switched hybridizing chain reaction (HCR). Ir-QAU was elaborately devised to present a signal-on response to G-quadruplex (G4) DNA against other DNAs due to its specific intercalation in G4 DNA and subsequent restriction of intra-molecular rotation. The recognition of H1 by Kan promoted the formation of Kan@H1 complexes, which hybridized with H2 and H3 via toehold-mediated hybridization reaction, subsequently switching HCR to produce large numbers of G4 DNA. Compared to Kan absence, abundant Ir-QAU was locked in G4 DNA to yield a significantly increased luminescence, which switches the luminescence analysis process of Kan with a limit of detection down to 0.38 pM. Furthermore, the Ir-QAU-based sensor was triumphantly applied to detect Kan in milk sample. We anticipate this work will disclose a new way to development of high-efficiency and practical luminescence sensor, and show a great potential for antibiotic-related food security.

Target-triggered dynamic hairpin assembly for signal amplification of microRNA and oncogenes and its application in live-cell imaging.

Based on target-triggered dynamic hairpin assembly (DHA) in both unidirectional and bilateral growth manners DNA nanobrushes are constructed, which realize sensitive and selective detection of short miRNA (miR-21) and long DNA (BRCA1), respectively. Moreover, the unidirectional DHA strategy is readily applied to in situ imaging of miR-21 in different live cells.

DNA logic assembly powered by a triplex-helix molecular switch for extracellular pH imaging.

We demonstrate a strategy for pH-programmable self-assembly of DNA nanostructures by taking advantage of the pH dependence of reverse Hoogsteen interactions for a triplex-helix molecular switch. This strategy is successfully applied to the construction of molecular logic gates and imaging of extracellular pH.

Isothermal exponential amplification techniques: From basic principles to applications in electrochemical biosensors.

As a conventional amplification technique, polymerase chain reaction (PCR) has been widely applied to detect a variety of analytes with exponential amplification efficiency. However, the requirement of thermocycling procedures largely limits the application of PCR-based methods. Alternatively, several isothermal amplification techniques have been developed since the early 1990s. In particular, according to the reaction kinetics, isothermal exponential amplification techniques possess higher amplification efficiency and detection sensitivity. The isothermal exponential amplification techniques can be mainly divided into two categories: enzyme-based isothermal exponential amplification and enzyme-free isothermal exponential amplification. Considering the advantages of high sensitivity and selectivity, high signal-to-noise ratio, low cost and rapid response time, exponential amplification electrochemical biosensors have attracted considerable attention. In this review, we introduce the basic principles of isothermal exponential amplification techniques and summarize their applications in electrochemical biosensors during the past five years. We also highlighted the present challenges and further perspectives of isothermal exponential amplification-based electrochemical biosensors.

Cross-catalytic hairpin assembly-based exponential signal amplification for CRET assay with low background noise.

A toehold-mediated strand displacement (TMSD)-based cross-catalytic hairpin assembly (C-CHA) is demonstrated in this study, achieving exponential amplification of nucleic acids. Functionally, this system consists of four hairpins (H1, H2, H3 and H4) and one single-stranded initiator (I). Upon the introduction of I, the first CHA reaction (CHA1) is triggered, leading to the self-assembly of hybrid H1·H2 that then initiates the second CHA reaction (CHA2) to obtain the hybrid H3·H4. Since the single-stranded region in H3·H4 is identical to I, a new CHA1 is initiated, which thus achieves cross operation of CHA1 and CHA2 and exponential growth kinetics. Interestingly, because the C-CHA performs in a cascade manner, this system can be considered as multi-level molecular logic circuits with feedback mechanism. Moreover, through incorporating G-quadruplex subunits and fluorescein isothiocyanate (FITC) in the product of H1·H2, this C-CHA is readily utilized to fabricate a chemiluminescence resonance energy transfer (CRET) biosensing platform, achieving sensitive and selective detection of DNA and microRNA in real samples. Since the high background signal induced by FITC in the absence of initiator is greatly reduced through labeling quencher in H1, the signal-to-noise ratio and detection sensitivity are improved significantly. Therefore, our proposed C-CHA protocol holds a great potential for further applications in not only building complex autonomous systems but also the development of biosensing platforms and DNA nanotechnology.