Dual-signal ratiometric electrochemiluminescence biosensor based on Au NPs-induced low-potential emission of PFO Pdots and LSPR-ECL mechanism for ultra-sensitive detection of microRNA-141.

In this study, we have for the first time constructed a ratiometric ECL biosensor for the ultrasensitive detection of microRNAs (miRNAs) using gold nanoparticles (Au NPs) to trigger both the low-potential emission from conjugated polymer poly(9,9-dioctylfluorene-2,7-diyl) dots (PFO Pdots) and the LSPR-ECL effect with sulfur-doped boron nitride quantum dots (S-BN QDs). PFO Pdots were first applied to the Au NPs-modified electrode, followed by covalent binding to capture the hairpin H1. Immediately thereafter, a small amount of miRNA-141 was able to generate a large amount of output DNA (OP) by traversing the target cycle. OP, H3-S-BN QDs, and H4-glucose oxidase (H4-GOD) were then added sequentially to the Au NPs-modified electrode surface, and the hybridization chain reaction (HCR) was initiated. This resulted in the introduction of a large amount of GOD into the system, which catalyzed the in situ formation of the co-reactant hydrogen peroxide (H2O2) from the substrate glucose. Due to the electron transfer effect, the production of H2O2 led to the ECL quenching of PFO Pdots. Meanwhile, H2O2 served as a co-reactant of S-BN QDs, resulting in strong ECL emission of S-BN QDs at the cathode. Furthermore, the cathodic ECL intensity of S-BN QDs was further enhanced by an LSPR-ECL mechanism between Au NPs and S-BN QDs. By measuring the ratio of ECL intensities at two excitation potentials, this approach could provide sensitive and reliable detection of miRNA-141 in the range of 0.1 fM ∼10 nM, with a detection limit of 0.1 fM.

An "off-on-enhanced on" electrochemiluminescence biosensor based on resonance energy transfer and surface plasmon coupled 3D DNA walker for ultra-sensitive detection of microRNA-21.

In this study, a novel electrochemiluminescence (ECL) biosensor was developed to detect microRNA-21 (miRNA-21) with high sensitivity by leveraging the combined mechanisms of resonance energy transfer (RET) and surface plasmon coupling (SPC). Initially, the glassy carbon electrode (GCE) were coated with Cu-Zn-In-S quantum dots (CZIS QDs), known for their defect-related emission suitable for ECL sensing. Subsequently, a hairpin DNA H3 with gold nanoparticles (Au NPs) attached at the end was modified over the surface of the quantum dots. The Au NPs could effectively quench the ECL signals of CZIS QDs via RET. Further, a significant amount of report DNA was generated through the action of a 3D DNA walker. When the report DNA opened H3-Au NPs, the hairpin structure experienced a conformational change to a linear shape, increasing the gap between the CZIS QDs and the Au NPs. Consequently, the localized surface plasmon resonance ECL (LSPR-ECL) effect replaced ECL resonance energy transfer (ECL-RET). Moreover, the report DNA was released following the addition of H4-Au NPs, resulting in the formation of Au dimers and a surface plasma-coupled ECL (SPC-ECL) effect that enhanced the ECL intensity to 6.97-fold. The integration of new ECL-RET and SPC-ECL biosensor accurately quantified miRNA-21 concentrations from 10-8 M to 10-16 M with a limit of detection (LOD) of 0.08 fM, as well as successfully applied to validate human serum samples.

Tripedal DNA Walker as a Signal Amplifier Combined with a Potential-Resolved Multicolor Electrochemiluminescence Strategy for Ultrasensitive Detection of Prostate Cancer Staging Indicators.

A multicolor electrochemiluminescence (ECL) biosensor based on a closed bipolar electrode (BPE) array was proposed for the rapid and intuitive analysis of three prostate cancer staging indicators. First, [Irpic-OMe], [Ir(ppy)2(acac)], and [Ru(bpy)3]2+ were applied as blue, green, and red ECL emitters, respectively, whose mixed ECL emission colors covered the whole visible region by varying the applied voltages. Afterward, we designed a simple Mg2+-dependent DNAzyme (MNAzyme)-driven tripedal DNA walker (TD walker) to release three output DNAs. Immediately after, three output DNAs were added to the cathodic reservoirs of the BPE for incubation. After that, we found that the emission colors from the anode of the BPE changed as a driving voltage of 8.0 V was applied, mainly due to changes in the interfacial potential and faradaic currents at the two poles of the BPE. Via optimization of the experimental parameters, cutoff values of such three indicators at different clinical stages could be identified instantly with the naked eye, and standard precision swatches with multiple indicators could be prepared. Finally, in order to precisely determine the prostate cancer stage, the multicolor ECL device was used for clinical analysis, and the resulting images were then compared with standard swatches, laying the way for accurate prostate cancer therapy.

Dual "on-off" signal conversion strategy based on surface plasmon coupling and resonance energy transfer for visual electrochemiluminescence ratiometric analysis of MiRNA-141.

An electrochemiluminescence (ECL) biosensor with a pair of new ECL emitters and a novel sensing mechanism was designed for the high-sensitivity detection of microRNA-141 (miRNA-141). Sulfur-doped boron nitrogen quantum dots (S-BN QDs) were initially employed to modify the cathode of the bipolar electrode (BPE), while the anode reservoir was [Ir(dfppy)2(bpy)]PF6/TPrA system. The next step involved attaching H1-bound ultra-small WO3-x nanodots (WO3-x NDs) to the S-BN QDs-modified BPE cathode via DNA hybridization. A strong surface plasmon coupling (SPC) effect was observed between S-BN QDs and WO3-x NDs, which allowed for the enhancement of the red and visible ECL emission from S-BN QDs. After target-induced cyclic amplification to produce abundant Zn2+ and Au NPs-DNA3-Au NPs (Au NPs-S3-Au NPs), Zn2+ could cleave DNA at a nucleotide sequence-specific recognition site to release the WO3-x NDs, resulting in the first diminution of cathode ECL signal and the first enhancement of anode ECL signal. Moreover, the ECL signal at cathode decreased for the second time and the emission of [Ir(dfppy)2(bpy)]PF6 was continuously enhanced after the introduction of Au nanoparticles-S3-Au nanoparticles on the cathode surface. Our sensing mode with a dual "on-off" signal conversion strategy shows a good detection capability for miRNAs ranging from 10-17 to 10-10 M, with a limit of detection (LOD) as low as 10-17 M, which has great application potential in biomedical research and clinical diagnosis.

Ultrasensitive and Visual Electrochemiluminescence Ratiometry Based on a Constant Resistor-Integrated Bipolar Electrode for MicroRNA Detection.

In this work, a new electrochemiluminescence (ECL) platform was constructed for detecting the prostate cancer marker microRNA-141 (miRNA-141) on a constant resistor-integrated closed bipolar electrode (BPE). It consisted of two reservoirs and a constant resistor, and both ends were connected to the anode of the driving electrode and the cathode of BPE. The cathode of BPE was modified with boron nitride quantum dots (BNQDs), and the anode reservoir was the [Ru(bpy)3](PF6)2/TPrA system. After introducing a certain amount of hairpin DNA 3 (H3) and ferrocene-labeled single-stranded DNA (Fc-ssDNA) on the surface of the BNQDs, the ECL emission signal of the BNQDs was difficult to be observed by the naked eye, while [Ru(bpy)3](PF6)2 emitted a strong and visible ECL signal. In the presence of the target, bipedal DNA assembled by catalytic hairpin assembly (CHA) took away the Fc-ssDNA and the ECL intensity of the BNQDs was enlarged, and as the concentration of miRNA-141 increased to the cutoff value, yellow-green light was visible by the naked eye. Meanwhile, the red emission signal of [Ru(bpy)3](PF6)2/TPrA became weakened. Thus, an ultrasensitive "color switch" ECL biosensor for detection of miRNA-141 was constructed and endowed with a wide linear range from 10-17 to 10-7 M and a detection limit of 10-17 M (S/N = 3). This study provides the potential for investigating portable devices in the detection of low-concentration nucleic acids.

Bipolar electrode ratiometric electrochemiluminescence biosensing analysis based on boron nitride quantum dots and biological release system.

In this article, we developed a novel ECL ratiometry on a closed bipolar electrode (BPE) for the sensitively and accurately detection of miRNA-21. High quantum yield and low toxicity BNQDs was synthesized and coated at BPE cathode as an ECL emitter, while the anode of BPE was calibrated via another ECL material, Ir(df-ppy)2(pic) (Firpic). The electron neutrality at both ends of the BPE electrically coupled the reactions on each pole of the BPE. Therefore, one electrochemical sensing reaction could be quantified at one end of the BPE. By the hybridization of target miRNA-21 and hairpin, the glucose blocked in MSNs by the hairpin was released and reacted with glucose oxidase (GOD) to generate H2O2, thereby reducing the ECL signal of the cathode BNQDs/K2S2O8 system and promoting ECL signal of anode Firpic/TPrA. Further, the G-quadruplex formed by unreacted hairpin bases consumed H2O2, which not only recovered the ECL of BNQDs, but also further improved the ECL emission of Firpic. Therefore, the concentration of miRNA-21 could be measured by the ECL ratio of BNQDs and Firpic. The data showed that the detection limit was 10-15 M (S/N = 3) with the linear range of 10-15 M to 10-9 M. The strategy of the BPE-ECL ratio method based on BNQDs showed a good prospect in clinical application.

Electrochemiluminescence Energy Resonance Transfer System between RuSi Nanoparticles and Hollow Au Nanocages for Nucleic Acid Detection.

This paper describes an electrochemiluminescence resonance energy transfer (ECL-RET) system using Ru(bpy)32+-doped silica nanoparticles (RuSi NPs) as the ECL donor and hollow Au nanocages as the ECL acceptor. Tetrahedron DNA (TD) was used to construct the biosensing interface and control the distance (4.8 nm) between the ECL donor-acceptor pairs. The surface plasmon resonance (SPR) nanostructures, Au nanocages were assembled via the hairpin based sandwich assay. Due to the well overlap between the plasmon absorption spectrum of Au nanocages (628 nm) and the ECL emission spectrum of RuSi NPs (620 nm), high efficient energy transfer could occur. Subsequent cyclic DNA amplification further increased the binding amount of Au nanocages. Since the ECL inhibition is closely related with the binding amount of Au nanocages, a general "signal-off" ECL bioassay could thus be tailored with high sensitivity. At the optimized conditions, this ECL-RET system performed well with great stability and repeatability for nucleic acid detection in the range from 1.0 fM to 10 pM. This work manifested the great promise of hollow Au nanocages for an ECL-RET biosensor that to the best of our knowledge has not been reported. We believe that it could inspire more interest in the design and development of numerous other SPR nanostructures for advanced ECL-RET biosensors.

Bidirectional Electrochemiluminescence Color Switch: An Application in Detecting Multimarkers of Prostate Cancer.

A selective excitation of [Ir(df-ppy)2(pic)] and [Ru(bpy)3]2+ through tuning the electrode potential is reported in this work. Bidirectional color change from blue-green to red could be observed along with increase and decrease of the potential, which was ascribed to the dual-potential excitation property of [Ir(df-ppy)2(pic)]. Similar to the three-electrode system, selective excitation of ECL could be achieved at the anode of the bipolar electrode (BPE). Both increase and decrease of the faradic reactions at the cathode of the BPE could induce ECL reporting color at the other pole switched from blue-green to red. We applied a closed BPE device for the bioanalysis of multicolor ECL since the organic solvent containing electrochemiluminophores could be separated from the bioanalytes. On the basis of BPE arrays coupled with the ECL switch, the detection of three biomarkers of prostate cancer, PSA, microRNA-141, and sarcosine were integrated in a same device. The cutoff values of the biomarkers could be recognized directly by the naked eye. Such a device holds great potential in the early diagnosis of prostate cancer.

Bipolar Electrode Based Multicolor Electrochemiluminescence Biosensor.

We report a multicolor ECL device based on closed bipolar electrode (BPE) for the visualized sensing of prostate-specific antigen (PSA) in human blood serum. As the emission color of concomitant electrochemiluminophores is potential resolved, similar to a three-electrode system, selective excitation of ECL could be achieved by tuning the interfacial potential at the poles of BPE. Via modulating the resistance of BPE, multicolor ECL emission of [Ru(bpy)3]2+ and [Ir(ppy)3] mixture using tripropylamine (TPrA) as the co-reactant was observed at the anode and the principle was elaborated. The concept was utilized to the quantification of clinical biomarkers with the color variation. A PSA concentration dependent silver bridge was constructed in the gap of the BPEs as an electric conductivity modulator. On the basis of a multicolor BPE-ECL device, the cutoff values (4.0 and 10.0 ng/mL) of human PSA could be recognized with naked eyes by the green-yellow-red ECL emission changing. As the first multicolor ECL device in biological analysis, BPE may raise the application of potential-resolved ECL to a new level.

Spatial-resolved electrochemiluminescence ratiometry based on bipolar electrode for bioanalysis.

Herein, a spatial-resolved electrochemiluminescene (ECL) ratiometry based on a closed biopolar electrode (BPE) is reported for the highly sensitive detection of prostate specific antigen (PSA). Au@g-C3N4 NCs as one ECL emitter were firstly coated on the cathode of BPE, while the anode of the BPE served for calibration via another ECL substance, Ru(bpy)3(2+). The electroneutrality across the BPE makes the reactions on each pole of BPE electrically coupled. Thus one electrochemical sensing reaction at one pole of BPE could be quantified at both ends. A composite, Pt-PAMAM-DNAzyme was assembled on the surface of cathode via DNA hybridization between probe DNA and PSA aptamer. It acted as an ECL quencher of g-C3N4 via resonance energy transfer (RET) and catalyzing the reduction of O2, the co-reactant of g-C3N4. Meanwhile, it could promote the ECL of Ru(bpy)3(2+) at anode, since the catalytic reduction of O2 at the cathode increased the faradiac current flowing through the BPE. Based on this signal composite, an ECL "off-on" phenomenon was observed at the cathode, after Pt-PAMAM-DNAzyme was "peeled off" by PSA. Conversely, at the anode, an "on-off" ECL changing was obtained. Therefore, a sensitive ratiometry for PSA detection was achieved with a linear range from 0.10 to 200ng/mL. Since the two ECL emitters were physically separated, the ratiometric system was relatively simple and neither optical filters nor spectrometer were required. The strategy combining the ECL ratiometry and BPE broadens the applications of BPE-ECL and shows good perspective in clinical application.

Visual Color-Switch Electrochemiluminescence Biosensing of Cancer Cell Based on Multichannel Bipolar Electrode Chip.

In this work, we developed a visual electrochemiluminescence (ECL) sensing platform based on a dual-bipolar electrode (D-BPE) array chip. The chip was composed of two arrays of BPEs and three separated arrays of reservoirs filled with buffer, Ru(bpy)3(2+)-TPrA and luminol solutions, respectively. Both BPEs served as ECL reporting platforms. By applying 6.0 V voltage, an array of orange electrochemiluminescence (ECL) signals belonging to Ru(bpy)3(2+) turned on. After adding DNAzyme and H2O2 in Ru(bpy)3(2+) and luminol reservoirs, the orange Ru(bpy)3(2+) signals decreased until vanished due to the quenching effect; meanwhile, a new array of blue ECL signals turned on because of the luminol-H2O2 ECL reaction. The designed D-BPE owns superior properties compared with the three-electrode system benefiting from the quantitative relation of bipolar systems, which greatly enhanced the ECL detection sensitivity. Meanwhile, the visual color-switch ECL and the ratiometric detecting principle made the results easier to evaluate and more accurate. Quantitative detection of HL-60 cancer cells from 320 cells/mL to 2.5 × 10(5) cells/mL with two linear ranges was achieved. The detection limit was down to 80 cells/mL. Finally, this D-BPE chip could distinguish the tumor cells from normal cells and provide a prospect for cancer diagnosis in a high-throughput, visual way.

A ratiometric electrochemiluminescence detection for cancer cells using g-C3N4 nanosheets and Ag-PAMAM-luminol nanocomposites.

In this work, a dual-signaling electrochemiluminescence (ECL) ratiometric sensing approach for the detection of HL-60 cancer cells was reported for the first time. G-C3N4 nanosheets and Ag-PAMAM-luminol nanocomposits (Ag-PAMAM-luminol NCs) were prepared and served as reductive-oxidative and oxidative-reductive ECL emitters respectively. DNA probe functionalized Ag-PAMAM-luminol NCs would hybridize with aptamers modified onto magnetic beads. In the presence of HL-60 cells, the aptamer would conjugate with the target cell and release Ag-PAMAM-luminol NCs. After magnetic separation, released Ag-PAMAM-luminol NCs would hybridize with capture DNA on g-C3N4 nanosheets. ECL from g-C3N4 nanosheets coated on ITO electrode at -1.25 V (vs SCE) could be quenched by Ag-PAMAM-luminol NCs due to the resonance energy transfer (RET) from g-C3N4 nanosheets to Ag NPs. Meanwhile, Ag-PAMAM-luminol brought the ECL signal of luminol at +0.45 V (vs SCE). Thus, the concentration of HL-60 cancer cells could be quantified by both the quenching of ECL from g-C3N4 nanosheets and the enhancement of ECL from luminol. By measuring the ratio of ECL intensities at two excitation potentials, this approach could achieve sensitive and reliable detection for cancer cells in a wide range from 200 cells/mL to 9000 cells/mL with the detection limit of 150 cells (S/N=3).

Visual electrochemiluminescence detection of telomerase activity based on multifunctional Au nanoparticles modified with G-quadruplex deoxyribozyme and luminol.

A novel visual electrochemiluminescence (ECL) analysis strategy for detection of telomerase activity is reported on a microarray chip, with G-quadruplex deoxyribozyme (DNAzyme) and luminol modified Au nanoparticles (NPs) as double-catalytic amplification labels.

Perovskite LaTiO3-Ag0.2 nanomaterials for nonenzymatic glucose sensor with high performance.

In this paper, a nonenzymatic glucose biosensor based on perovskite LaTiO3-Ag0.2(LTA) modified electrode was presented. The morphology and the composition of the perovskite LaTiO3-Ag0.2 nanomaterials were characterized by using scanning electron microscopy (SEM) and X-ray diffraction (XRD) respectively. The LaTiO3-Ag0.2(LTA) composite was investigated by electrochemical characterization using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimal conditions, CV and chronoamperometry (I-t) study revealed that, compared with the bare glassy carbon electrode (GCE), the modified electrode showed a remarkable increase in the efficiency of the electrocatalytic oxidation of glucose, starting at around +0.70 V (vs. Ag/AgCl). The prepared sensor exhibited a high sensitivity of 784.14 µAmM⁻¹ cm⁻², a low detection limit of 2.1×10⁻7 M and a wide linear range from 2.5 µM to 4 mM (R=0.9997). More importantly, the LTA modified electrode was also relatively insensitive to commonly interfering species such as ascorbic acid (AA), uric acid (UA), dopamine (DA) in high potential. Moreover, the nonenzymatic sensor was applied to the determination of glucose in human serum samples and the results were in good agreement with clinical data. Electrodes modified with perovskite nanomaterials are highly promising for nonenzymatic electrochemical detection of glucose because of their high sensitivity, fast response, excellent stability and good reproducibility.