Enhanced Electrochemiluminescence Efficiency Research Articles

Direct Comparative Studies Revealing the Contribution of TADF Activity of Organic Emitters Towards Efficient Electrochemiluminescence.

Electrochemiluminescence (ECL) featuring thermally activated delayed fluorescence (TADF) properties has attracted considerable interest, showcasing their potential for 100 % exciton harvesting, which marks a significant advancement in the realm of organic ECL. However, the challenge of elucidating the precise contribution of TADF to the enhanced ECL efficiency arises due to the lack of comparative studies of organic compounds with or without efficient TADF properties. In this study, we present four carbazole-benzonitrile molecules possessing similar chemical structures and comparable exchange energy (ΔEST). Despite their comparable properties, these compounds exhibited varying TADF efficiencies, warranting a closer examination of their underlying structural and electronic characteristics governing the optical properties. Consequently, intense ECL emission was only observed from 4CzBN with a remarkable TADF efficiency, underscoring the substantial difference in the ECL signal among molecules with comparable ΔEST and similar spectral properties but varying TADF activity.

Supramolecular assembly-induced electrochemiluminescence enhancement of gold nanoclusters for hemoglobin detection.

Gold nanoclusters (Au NCs) with excellent optical properties and biocompatibility have become one of the most promising electrochemiluminescence (ECL) emitters. However, the low efficiency and poor stability of Au NCs restrict their applications in ECL. Herein, by supramolecular assembly of L-arginine (Arg) and 4-hydroxy-2-mercapto-6-methylpyrimidine (MTU) on the surface of Au NCs, Arg/MTU-Au NCs with enhanced ECL efficiency and stability were prepared. Compared with the MTU-stabilized Au NCs (MTU-Au NCs), the ECL efficiency of Arg/MTU-Au NCs increased by 24.8 times. As a proof-of-concept, a sensitive biosensing platform was constructed for sensitive detection of hemoglobin (Hb) in urine using Arg/MTU-Au NCs as ECL emitters. The proposed ECL detection platform provides a feasible strategy for the detection of biomarkers in urine and has broad application prospects in disease screening and clinical marker detection.

Electrochemiluminescence resonance energy transfer between Ru(bpy)32+@Cu3(HHTP)2 and GO-Au composites for C-reactive protein detection

Designing innovative electrochemiluminescence (ECL) immunosensors is critical for the detection of biomarkers with a low concentration and the precise evaluation of clinical diseases. Herein, a Cu3(hexahydroxytriphenylene)2 (Cu3(HHTP)2) nanoflake-based sandwich-type ECL immunosensor was constructed for C-Reactive Protein (CRP) detection. The Cu3(HHTP)2 nanoflake, an electronically conductive metal–organic framework (MOF), has a periodically arranged porous structure with a cavity size of 2 nm, which not only accommodates a large amount of Ru(bpy)32+ but also confines the spatial diffusion of active species. Therefore, the Ru(bpy)32+-loaded Cu3(HHTP)2 nanocomplex (Ru@CuMOF) as an ECL emitter exhibits an enhanced ECL efficiency. The ECL resonance energy transfer (ECL-RET) was accomplished by combining Ru@CuMOF used as a donor with gold nanoparticles-functionalized graphene oxide nanosheets (GO-Au) utilized as an acceptor. This should be ascribed to the fact that the ECL emission spectrum of Ru@CuMOF shows the strongest signal intensity at 615 nm, overlapping with the absorption spectrum of GO-Au at 580–680 nm. Targeted detection of CRP in human serum samples was achieved by the sandwich-type immunosensor based on the ECL-RET mechanism with a 0.26 pg mL−1 detection limit. The electro-activated hybrids of Cu3(HHTP)2 and ECL emitters provide a new sensing strategy for the high-sensitivity detection of disease markers.

Schiff Base Complexes with Covalently Anchored Luminophores: Self-Enhanced Electrochemiluminescence Detection of Neomycin.

A novel electrochemiluminescence (ECL) amplification strategy was established aiming to overcome the inherent shortcomings of the current oxygen (O2) coreactant ECL systems. Macrocyclic Schiff base Fe complexes were rationally designed as a novel integrated ECL emitter by iminium linkage between N-(4-aminobutyl)-N-ethylisoluminol (ABEI) and 1,10-phenanthroline-2,9-dicarbaldehyde (PDL) and postmetalation of the macrocyclic Schiff base. Covalently combining luminophore ABEI with a catalytic center endowed the novel ECL emitter with both remarkable redox electrocatalytic properties and significantly enhanced ECL efficiency. The high content of ferrous iron and the dominantly active low-spin Fe state greatly contributed to the inherent catalytic activity for O2 activation. The rational modification of luminophore optimized the spatial distribution and simultaneously shortened the species transport distance of coreactant radicals generated in situ from dissolved O2, resulting in significantly self-enhanced ECL efficiency. Neomycin, which posed a growing threat to aquatic biodiversity and environmental safety, as the model antibiotic was successfully detected with a detection limit of 0.21 pM (S/N = 3), clarifying a promising application prospect of this new luminophore-embedded ECL amplification strategy in biological analysis and environmental monitoring.

Covalent organic framework linked with amination luminol derivative as enhanced ECL luminophore for ultrasensitive analysis of cytochrome c.

Cytochrome c (cyt c) plays a critical role in mitochondrial respiratory chain, whose absence is detrimental to electron transport and reduce adenosine triphosphate. For ultrasensitive detection of cyt c, sheet-like covalent organic frameworks (COFs) were prepared by orderly accumulation of 1,3,5-benzenetricarboxaldehyde (BTA) and p-phenylenediamine (PDA), and further grafted with N-(4-aminobutyl)-N-ethylisoluminol (ABEI) - an electrochemiluminescence (ECL) emitter. Specifically, the morphology and structure of the COFs-ABEI were mainly characterized by transmission electron microscopy (TEM), X-ray diffraction (XRD) analysis, and X-ray photoelectron spectroscopy (XPS). In parallel, the optical properties of the emitter were certified by UV-vis absorbance spectroscopy, Fourier infrared spectroscopy (FTIR), fluorescence (FL), and ECL measurements, showing 2.25-time enhanced ECL efficiency over pure ABEI, coupled by illustrating the interfacial electron transport mechanism. On the above foundation, a label-free "signal off" ECL biosensor was constructed by virtue of the specific immune recognition between the aptamer of the target cyt c with its capture DNA (cDNA) anchored on the biosensing platform, exhibiting a wider linear range of 1.00 fg mL-1-0.10 ng mL-1 (R2 = 0.998) and a lower limit of detection (LOD) down to 0.73 fg mL-1. This work offers some constructive guidelines for sensitive bioassays of disease-related biomarkers in the clinical field.

Quadrilateral Nucleic Acid Frame-Accelerating DNAzyme Walker Kinetics for Biosensing Based on Host-Guest Recognition-Enhanced Electrochemiluminescence.

Depending on the reaction between walkers and tracks, DNA walker is able to output signals continuously, which has attracted great attention from the bioanalytical community. Therefore, how to improve its reaction kinetics for efficient signal readout is of great significance. Herein, a quadrilateral DNAzyme walker was fabricated by colocalizing one walker and three DNA tracks in the quadrilateral nucleic acid frame to form a reaction unit (abbreviated as qDNA walker). Impressively, in contrast to the common free DNAzyme walker, the reaction kinetics of the qDNA walker was 2.3 times faster, which could achieve microRNA detection within 30 min. Meanwhile, an electrochemiluminescence (ECL) emitter of anthracene-cucurbituril supramolecular nanocrystals (Ant-CB SNCs) was obtained based on the self-assembly of cucurbituril (CB, host molecule) and anthracene (Ant, guest molecule). Benefiting from the host-guest recognition effect, the prepared Ant-CB SNCs exhibited enhanced ECL efficiency due to the supramolecular interaction between CB and Ant, which could inhibit vibration and rotation of the Ant molecules. We defined this new enhanced ECL phenomenon as "host-guest recognition-enhanced ECL." As a proof of concept, an ECL biosensor for microRNA-21 (miRNA-21) was constructed by combining the high-efficiency DNAzyme walker and the advanced ECL emitter of Ant-CB SNCs, which showed a linear range from 50 aM to 50 pM with a low limit of detection (11 aM), highlighting the great potential in clinical diagnosis.

Aggregation-Induced Electrochemiluminescence of Tetraphenylbenzosilole Derivatives in an Aqueous Phase System for Ultrasensitive Detection of Hexavalent Chromium.

Herein, aggregation-induced electrochemiluminescence (AIECL) of tetraphenylbenzosilole derivatives in an aqueous phase system with the participation of a co-reactant was systematically investigated for the first time. All organics that we studied exhibit excellent stability and dramatically enhanced electrochemiluminescence (ECL) and photoluminescence (PL) emission when the water fraction increases. The influence of substituents in the structure of tetraphenylbenzosilole derivatives on AIECL performance was proved by fluorescence, cyclic voltammetry, and related theoretical calculation. Among them, 2,3-bis(4-cyanophenyl)-1,1-diphenyl-benzosilole (TPBS-C) with strong electron-withdrawing cyano groups exhibits the best ECL behavior with the highest ECL efficiency (184.36%). The strongest ECL emission of TPBS-C not only stems from the aggregated molecules that restrict the intramolecular motion of peripheral phenyl groups, which inhibits the nonradiative transition, but also comes from the fact that TPBS-C has the lowest reduction potential, and twice the reduction process of TPBS-C occurs to produce more anion radicals (TPBS-C·-). Significantly, the ECL sensor based on TPBS-C nanoaggregates exhibits excellent detection performance for toxic Cr(VI) with a wide linear range from 10-12 to 10-4 M and an extremely low detection limit of 0.83 pM. This work developed an efficient luminophore with unique AIECL properties and realized the ultrasensitive detection of Cr(VI) in the aqueous phase system.

In Situ Controllable Generation of Copper Nanoclusters Confined in a Poly-l-Cysteine Porous Film with Enhanced Electrochemiluminescence for Alkaline Phosphatase Detection.

Copper nanoclusters (Cu NCs) as emerging luminescent metal NCs are gaining increasing attention owing to the comparatively low cost and high abundance of the Cu element in nature. However, it remains challenging to manipulate the optical properties of Cu NCs. Unlike most dispersed Cu NCs, whose luminescence efficiency was restricted by nonexcited relaxation, the Cu NCs confined in a porous poly-l-cysteine (poly-l-Cys) film were generated controllably with enhanced electrochemiluminescence (ECL) by in situ electrochemical reduction. Specifically, poly-l-Cys provided a porous structure to regulate the generation of Cu NCs within its holes, which not only increased the restriction on the intramolecular vibration and rotation of the ligands but also expedited the electron transfer near the electrode surface, reflecting in an enhancement of the ECL signal and efficiency. As an application of the confined Cu NCs, an ECL biosensor with high performance was constructed skillfully for highly sensitive detection of alkaline phosphatase (ALP), which adopted Cu NCs as the ECL luminophore and poly-l-Cys as a coreaction accelerator in a novel ECL ternary system (Cu NCs/S2O82-/poly-l-Cys). Furthermore, an ingenious target amplification based on the combination of a DNA walker and click chemistry was developed to convert ALP to DNA strands efficiently, achieving great improvement in the recognition efficiency. As a result, the biosensor had a low detection limit (9.5 × 10-7 U·L-1) and a wide linear range (10-8-10-2 U·L-1) for ALP detection, which showed great promise for the detection of non-nucleic acid targets and the diagnosis of diseases.

Oxygen Vacancy-Enhanced Electrochemiluminescence Sensing Strategy Using Luminol Thermally Encapsulated in Apoferritin as a Transducer for Biomarker Immunoassay.

Oxygen vacancies (OVs) enhanced electrochemiluminescence (ECL) biosensing strategy using luminol thermally encapsulated in apoferritin (Lum@apoFt) as an efficient transducer was investigated for ultrasensitive biomarker detection. By applying the oxygen-defect engineering (ODE) strategy, the OVs enriched cobalt-iron oxide (r-CoFe2O4) was fabricated as the sensing substrate to boost the electron mobility and catalyze the generation of superoxide anion radical (O2•-) for signal amplification. It should be noted that r-CoFe2O4 with higher OVs density dramatically accelerated the ECL reaction between O2•- and luminol anionic radicals, achieving 6.5-fold stronger ECL output than CoFe2O4 with no or low OVs density. Moreover, facile encapsulation of approximate 412 luminol molecules in a single apoFt cavity was first realized by an efficient thermal-induction method. The obtained Lum@apoFt complexes exhibited well-maintained ECL efficiency and excellent biocompatibility for biological modifications. On this basis, a biosensor was developed for early diagnostics of squamous cell carcinomas by detecting its representative biomarker named cytokeratin 19 fragment 21-1 (CYFRA 21-1), from which excellent linearity was achieved in 0.5 pg/mL to 50 ng/mL with a detection limit of 0.14 pg/mL. This work not only put forward a novel idea of creating OVs enriched sensing interface with excellent signal-amplification function but also proposes a facile and robust methodology to design apoFt-based transducers for developing more practical nanoscale biosensors in early diagnostics of diseases.

A three-dimensional DNA nanomachine with target recycling amplification technology and multiple electrochemiluminescence resonance energy transfer for sensitive microRNA-141 detection

An electrochemiluminescence (ECL) three-dimensional (3D) DNA nanomachine is developed for microRNA-141 (miRNA-141) detection by coupling Pb2+ dependent DNAzyme assisted target recycling amplification technology with multiple ECL resonance energy transfer (ECL-RET) system. Firstly, Pb2+ dependent DNAzyme is formed by three single strand DNA (ssDNA): A1, A2 and the target miRNA-141. In the presence of Pb2+, the specific recognition site of the DNAzyme is cleaved and a large number of secondary targets (A3) are released. Secondly, the 3D DNA nanomachine consists of four ssDNA: H1, H2, H3 and the probe (two ends are labeled with alexa fluor (AF) and a nanocomposite (PtNCs@Ru(dcbpy)32+) which is prepared by polyethyleneimine platinum nanoclusters and tris(4,4′-dicarboxylicacid-2,2′-bipyridyl) ruthenium(II) dichloride). Then, the 3D DNA nanomachine is assembled on the gold nanoparticles modified glassy carbon electrode. Afterwards, A3 is employed to hybridize with the probe, triggering the movement of the nanomachine and forming the multiple ECL-RET system. In this system, AF, serves as an effective energy transfer donor, which can transfer energy to PtNCs and Ru(dcbpy)32+directly. Meanwhile, PtNCs, both as the acceptor and donor, can accept energy from AF and transfer it to Ru(dcbpy)32+. As a result, The biosensor achieves enhanced ECL efficiency, which is 1.78 times that of the classic tris(2,2′-bipyridyl)ruthenium(II) dichloride (Ru(bpy)32+) and exhibits good responses to miRNA-141 in the linear range from 10 aM to 100 nM with a detection limit of 3.3 aM. Also, the obtained biosensor can be employed to detect miRNA-141 in human serum samples, which will be of great significance in bioanalysis.

Restriction of intramolecular motions (RIM) by metal-organic frameworks for electrochemiluminescence enhancement:2D Zr12-adb nanoplate as a novel ECL tag for the construction of biosensing platform

Herein, a new phenomenon of enhanced electrochemiluminescence (ECL) emission by restricting intramolecular motion in the 2D ultra-thin Zr12-adb (adb = 9,10-anthracene dibenzoate) metal-organic framework (MOF) nanoplate was discovered for the first time. The coordination immobilization of adb in porous ultra-thin Zr12-adb nanoplate endowed the Zr12-adb excellent ECL performance, including stronger ECL signal and higher ECL efficiency relative to those of H2adb monomers and H2adb aggregates. In the 2D Zr12-adb nanoplate, the bridging ligand adb was stretched and fixed between two Zr12 clusters, which restricted intramolecular rotations and suppressed unnecessary energy loss caused by self-rotation, thereby remarkably improved the ECL intensity and efficiency. More importantly, the porous ultra-thin structure of Zr12-adb MOF nanoplate not only allowed the coreactants to diffuse into the MOF interior, making both internal and external adb be excited, but also shortened the migration distance of electrons, ions, coreactants and coreactant intermediates, which further improved the ECL efficiency of Zr12-adb and overcame the shortcoming of H2adb aggregates in which the internal luminophores were not easily excited. Regarding the excellent ECL properties above, Zr12-adb nanoplate was selected as a new ECL emitter incorporated with the bipedal walking molecular machine together to fabricate a biosensor for sensitive detection of mucin 1. The enhanced ECL by restriction of intramolecular motions in MOFs provided a new pathway to improve ECL intensity and efficiency, which lighted up a lamp for the design and manufacture of high-performance ECL materials based on MOFs, thus offering new opportunities to develop ultrasensitive ECL biosensors.

Ultrasensitive electrochemiluminescence biosensor for organophosphate pesticides detection based on carboxylated graphitic carbon nitride-poly(ethylenimine) and acetylcholinesterase

A signal on electrochemiluminescence (ECL) biosensor was designed for sensitive organophosphate pesticides (OPs) detection based on a novel composite of carboxylated graphitic carbon nitride-poly(ethylenimine) (C-g-C3N4-PEI) and acetylcholinesterase (AChE). The C-g-C3N4-PEI nanocomposite which was prepared through covalent bonding between the COOH of C-g-C3N4 and the NH2 of PEI exhibited significantly enhanced ECL efficiency and stability. K2S2O8 as the coreactant of C-g-C3N4-PEI could be consumed by thiocholine, produced by the hydrolysis of acetylthiocholine (ATCl) in the presence of AChE. Since OPs are one of AChE inhibiter, the consumption of coreactant K2S2O8 decreased with the increasing concentration of OPs, thus enhancing ECL signal. Under the optimum conditions, the proposed biosensor for OPs detection (using ethyl paraoxon as a model OPs) exhibited a wide linear ranging from 1.0 pM to 5.0μM with a low detection limit of 0.3 pM. The novel strategy has the advantages of fine practicality, good stability and reproducibility, which might provide a new promise for OPs detection in real-life samples.

Target-catalyzed hairpin assembly and intramolecular/intermolecular co-reaction for signal amplified electrochemiluminescent detection of microRNA.

Herein, a new electrochemiluminescence (ECL) strategy for enzyme-free microRNA-21 (miR-21) amplified detection was designed based on target-catalyzed hairpin assembly by combining the signal-amplification capability of both intramolecular and intermolecular ECL co-reaction. In this strategy, two hairpin DNA probes of H1 and H2 were designed as capture probes and detection probes, respectively. To be specific, the capture probes of H1 were immobilized on the multilayer interface of AuNPs and thiosemicarbazide (TSC) assembly on the single-walled carbon nanohorns decorated electrode, while the detection probes of H2 was anchored on the nanocarriers of gold nanoparticals functionalized reduced graphene oxide (Au-rGO) which were tagged with the self-enhanced ruthenium complex (PEI-Ru(ΙΙ)) in advance. Based on the target-catalyzed hairpin assembly, target miR-21 could trigger the hybridization of H1 and H2 to further be released for initiating the next hybridization process to capture a large number of H2 bioconjugates on the sensing surface. Herein, the TSC was used not only as a coupling reagent to attach the AuNPs via Au-S and Au-N bonds but also as a novel intermolecular coreactant to enhance the ECL intensity, and the PEI-Ru(ΙΙ) as emitters exhibited enhanced ECL efficiency. Therefore, a strong ECL signal was achieved by the dual amplification strategies of target recycle and the intramolecular/intermolecular co-reaction of PEI-Ru(ΙΙ) and TSC. The designed protocol provided an ultrasensitive ECL detection of miR-21 down to the sub-femtomolar level with a linear response about 6 orders of magnitude (from 1.0 × 10(-16)M to 1.0 × 10(-11)M) with a relatively low detection limit of 0.03 fM (S/N=3).

Ultrasensitive Apurinic/Apyrimidinic Endonuclease 1 Immunosensing Based on Self-Enhanced Electrochemiluminescence of a Ru(II) Complex

An alternative "signal on" immunosensor for ultrasensitive detection of apurinic/apyrimidinic endonuclease 1 (APE-1) was designed utilizing the self-enhanced electrochemiluminescence (ECL) of a novel Ru(II) complex functionalized coil-like nanocomposite as signal labels. The desirable self-enhanced ECL luminophore was achieved by combining the coreactant of poly(ethylenimine) (PEI) and the luminophor of bis(2,2'-bipyridine)-5-amino-1,10-phenanthroline ruthenium(II) [Ru(bpy)2(5-NH2-1,10-phen)(2+)] to form one novel Ru(II) complex, which exhibited significantly enhanced ECL efficiency and stability. Moreover, the carbon nanotubes (CNTs) were employed as nanocarriers for self-enhanced Ru(II) complex loading via π-π stacking to obtain the coil-like nanocomposite to act as signal probe. Compared with traditional ECL immunoassay, our proposed strategy is simple and sensitive, avoiding the adding of any coreactant into testing solution for signal amplification, and shows a detection limit down to subfemtogram per milliliter level under the optimized experimental condition.

The enhanced electrochemiluminescence of lucigenin by some hydroxyanthraquinones

Several hydroxyanthraquinones (emodin, rhein and physcion) were found to strongly enhance the cathodic electrochemiluminescence (ECL) of the lucigenin by scanning under the mode of differential pulse voltammetry. The enhanced intensity was linear with the concentrations of rhein, emodin and physcion. The linear calibration ranges of 8.0 × 10 −8 to 2.0 × 10 −6, 3.0 × 10 −7 to 8.0 × 10 −6 and 1.0 × 10 −7 to 1.0 × 10 −6 mol/L, the detection limits of 2.1 × 10 −8, 1.6 × 10 −7 and 5.2 × 10 −8 mol/L were obtained for rhein, emodin and physcion, respectively. Potential-resolved ECL was used to study the possible mechanism of the enhancement effect. An electron transfer pathway was found to be involved in the ECL process. It has been confirmed that the reduced hydroxyanthraquinones reacted with dioxygen to give superoxide radical anion which increased the ECL of the lucigenin. Furthermore, these three hydroxyanthraquinones revealed the different enhanced ECL efficiency in the following order: rhein > physcion > emodin.