A Review on the Progress of Metal-Organic Frameworks in Electrochemiluminescence Sensors
A Review on the Progress of Metal-Organic Frameworks in Electrochemiluminescence Sensors
Electrogenerated chemiluminescence, also called ECL, is a kind of light emission process.[1, 2] It is a production of electrochemical and chemiluminescence (CL) reactions at the surface of electrode. ECL mechanisms are mainly classified into two classes: coreactant mechanism and annihilation mechanism. The former is mostly studied in the research. The ECL is induced by applying voltage or current in a system that contains luminescent and coreactant substance, and new species are generated by the electrochemical reaction of coreactant. Then the new species would react with the luminescent substance and undergo highenergy electron-transfer reactions to produce a light-emitting excited state of luminescent substance, finally the excited state of luminescent substance returns to the ground state and releases light. Compared with CL, the tempus and spatiality of ECL are easily controllable by triggering or pausing the electrochemical reaction. Compared with photo-luminescence, ECL does not suffer from background interference, because no light source is needed. By combining the electrochemical and optical merits, ECL exhibits several fantastic advantages such as excellent sensitivity, fast response, broad and dynamic scope, low background, and simplicity. Due to these benefits, ECL sensors, which are defined as using active molecules as molecular recognition elements and ECL reagents as signal substances to measure the concentration of target molecules by translating a chemical interaction into a quantifiable ECL signal, have been used for a widespread application from experimental to the commercial field.[3, 6] However, with further study of biomolecules, the high requirements for ECL sensors are put forward. In addition to high sensitivity and high specificity, precise detection, in-situ detection, real-time detection, ultra-trace detection, and others attract more and more attention.
MOFs, composed of metal ions or clusters and organic ligands via coordination bonds, are a class of hybrid porous materials with high crystallinity.[7-9] Since first discovery in 1989 by Hoskins and Robson, as the high porosity and possibility for suitably tuning both their properties and structure on demand, [9, 11-17] MOFs are regarded as a promising material for application in a wide range of fields, including gas adsorption and separation, [18-20] energy storage, [21-23] biomedicine,  catalysis, [25-27] and sensors.[28-34] In ECL sensors domain, the tunable properties of MOFs carried out high diversity in the application, which has been used in the construction of sensing interfaces for assembling abundant molecular recognition elements to capture more analytes,  serving as ECL signal emitter material or signal material carriers,  as the co-reaction accelerator to promote the reaction rate between ECL emitter and coreactant or active elements in mimic enzymes,  to cover the high requirements for testing. However, as research on ECL sensors has progressed in width and depth, the shortcomings of MOFs become apparent, e.g. poor conductivity, low water stability, and weak biocompatibility. To address these drawbacks, lots of efforts have been devoted to remedying the performance of MOFs in ECL sensors by altering the synthesis method/condition,  synthesizing new kinds of MOFs,  proper modifying/functionalizing the pristine MOFs,  or deriving MOFs to other nanomaterials. The new generation of MOFs with sufficiently attractive characteristics is extensively applied in ECL sensors recently, followed by the charge or energy transfer progress occurring in MOFs under ECL stimulation which is blooming proposed, such as ligand-to-metal charge transfer (LMCT) progress,  and ligand-to-ligand charge transfer (LLCT) progress. And the fact should arouse our attention that MOFs contain so various characters as to render their modulation to themselves ECL performance relatively more complicated in the application of ECL sensors. Therefore, the summary and analysis of the relationship between characters of MOFs and their ECL performance would point out the light for the rational construction of new MOFs-based ECL sensors.
In this review, we emphasize describing the applications of MOFs in the domain of ECL sensors for the detection of various targets, including metal ions, small molecules, nucleic acids, proteins, bacteria, and viruses. Then, the relationship between ECL performance and the characters of MOFs is briefly discussed in the representative examples. At last, we provide the potential opportunities and challenges faced by MOFs in the realm of ECL sensors, as well as the future perspectives. Owing to the explosion of papers in this active field, we certainly missed many important contributions, so we apologize to the authors in advance that is unintentionally left out.
MOFS IN ECL SENSORS
Metal Ions Sensing. Metal ions are universal in daily life and closely related to human health and environment. The process of human metabolism is inevitable from metal ions, and overdosed or lack of these ions will lead to various diseases. As a result, the detection of metal ions is an important issue in human health. Mercury (Hg) is a heavy metal element with a significant accumulation effect and genotoxicity, mainly existing in the form of mercury ions (Hg2+) in water. A "signal-on" ECL sensor for sensitive detecting Hg2+ is established on the basis of that the tris(bipyridine)ruthenium(II)-functionalized metal-organic frameworks (Ru-MOFs) can be disassembled and release tris(bipyridine)ruthenium(II) (Ru(bpy)32+) in the response of Hg2+ ion (Figure 1). The solvothermal method is used to prepare Ru-MOFs by adding and heating adenine, 4, 4′-biphenyl dicarboxylic acid (BPDC), and zinc nitrate hexahydrate in the mixed solvent. The obtained rod-shaped crystals are Ru-MOFs. Ru(bpy)32+ ions occupy the pores and dope the frameworks as BPDC in the structure of Ru-MOFs, while the interaction between Ru(bpy)32+ and negatively charged MOFs is electrostatic interaction rather than bonding. In the presence of Hg2+, adenine in the solution can strongly and selectively integrate with Hg2+, leading to an increasing positive charge near Ru(bpy)32+. Then, Ru(bpy)32+ releases out from the framework by electrostatic repulsion. With the massive Ru(bpy)32+ released, the structure of MOFs collapsed, even decomposed. The ECL intensity is linearized with the concentration of Hg2+ in the range of 1-100 pM, with a detection limit of 0.53 pM. Since the smartphone-based detection is able to meet the requirements of on-site tests, a colorimetric and ECL dual model analytical platform is fabricated for visual and sensitive detection of Hg2+ by using two thymine (T)-rich DNA probes as capture probes and alkaline phosphatase (ALP) to catalyze peroxydisulfate/metal-polydopamine frameworks@Ru (S2O82-/MPFs@Ru) system. By comparing the two above methods, the use of a DNA capture probe ensures the specificity of the method, and the integration of a smartphone is expected to achieve rapid on-site determination.
Pb(II) ions (Pb2+), as one of the heavy metal ions, can inactivate enzymes and have great toxicity to human tissue. The detection of Pb2+ is of great importance, many researchers have investigated the MOFs-based ECL sensors for detecting Pb2+. An ECL sensor for detecting Pb(II) is constructed based on a Ru(bpy)32+ doped UiO-66 Zr-MOFs and an amino-functionalized silica nanosphere. With the large specific surface area and high porosity of UiO-66, the loading efficiency of Ru(bpy)32+ is increased and ECL intensity significantly enhanced, the sensitivity improved further, and the ultrasensitive detection limit is 1.0 × 10-7 μM. Other than the above-mentioned single metal ion cases, MOFs-based ECL sensors for multiple determination of metal ions have also been reported. For instance, cadmium selenide quantum dots hybridized MIL-53(Al) composite (MIL-53(Al) @CdTe QDs) and gold nanoparticles (Au NPs) or platinum nanoparticles (Pt NPs) label-aptamer are used to fabricate an aptasensor for Pb2+ and Hg2+ determination. MIL-53(Al)@CdTe QDs composite is used to modify glassy carbon electrode (GCE) to improve the electrical conductivity of GCE, and as an ECL signal material to measure the concentration of Pb2+ and Hg2+ by the interaction of enhanced resonance energy transfer (ERET) and attenuated surface plasmon resonance (SPR) between AuNPs/PtNPs and MIL-53(Al)@CdTe QDs.
MOFs-based ECL sensors can also be extended to probe other metal ions such as hexavalent chromium (Cr6+), cooper(II) (Cu2+), and ferrous(II) (Fe2+). For example, Ma et al. utilized the collisionally ECL quenching effect of Cr6+ to ECL emitter (gold nanoparticles-hybridized Pb(II)-β-cyclodextrin metal-organic framework (Au@Pb-β-CD) composites) for the ultrasensitive determination of Cr6+ in river water (Figure 2). The excellent ECL behavior of Au@Pb-β-CD composites mainly subscribed from Pb-β-CD and the enhancement of Au NPs. Hu et al. designed a dual-modality readout sensing platform for analyzing Cu2+, coenzyme, and histone acetyltransferase based on an electrosynthesized Ru-MOFs. In addition, based on the inhibition effect of ferrous ion on the ECL emitter of copper(II) metalorganic framework ([Cu(L)(H2O)2]nCu-MOF), a highly enhanced ECL sensor is constructed for Fe2+ detection.
Conventionally, the role of MOFs in the ECL detection of metal ions can be categorized as the construction of sensing interfaces to enhance the electrical conductivity of the sensing interface, as ECL signal emitter material because a dramatic change of ECL signal would bring out by the reaction between target with MOFs, or signal material carriers to improve the sensitivity of detection.
Small Molecules Sensing. In a regular physical checkup, small molecules such as dopamine (DA), ascorbic acid (AA), uric acid (UA), and glucose in human blood or urine are important physiological indexes, so accurate and rapid determinations of these small molecules are significant for the evaluation of various diseases. Recently, on the basis of quenching effects, DA, an important neurotransmitter, has been sensitively detected by MOFs-based ECL sensors. An "on-off" ECL sensor based on the dual molecular recognition strategy and the quenching effect of Fe-MIL-88 MOFs on the ECL behavior of 3, 4, 9, 10-perylenetetra-carboxylic acid-hydrogen peroxide (PTCA-H2O2) system has been constructed by Fu et al. Dual molecular recognition consists of two recognition mechanisms. One is that the 3-aminopropyltri-ethoxysilane functionalized PTCA modified GCE (APTES-PTCA/GCE) specifically recognizes and immobilizes target DA via the diol from DA and the oxyethyl group from APTES, and the other is that target DA captures quencher probe Fe-MIL-88 MOFs via amide interaction. In the presence of DA, Fe-MIL-88 MOFs are captured onto the surface of APTES-PTCA/GCE and the resonance energy transfer (ECL-RET) from PTCA to Fe-MIL-88 MOFs is activated, resulting in ECL quenching of PTCA. The ECL intensity of APTES-PTCA/GCE is decreased with the increasing concentration of DA, indicating an "on" to "off" MOFs-based ECL sensor for DA. In another report, Ru(bpy)32+ functionalized Zn(II) metal-organic frameworks are designed as efficient ECL emitters in ECL sensor for DA detection. In the case of ECL detection of DA, the ECL intensity of Ru-MOFs is quenched by the oxidation products of DA and charge/energy transfer between DA and ECL emitters. The MOFs with large internal surface areas encapsulate abundant Ru(bpy)32+ to effectively prevent the leakage of Ru molecules, which results in high stability of ECL sensors.
AA is popular for its antioxidant property, which requires in the daily diet and is associated with fertility. A Zn-based porphyrin-based metal-organic framework ((Zn)porphMOF) is used as an efficient ECL emitter to detect AA based on the fact of the dramatic confinement of AA to the energy and electron transfer of (Zn)porphMOF. Meanwhile, an exogenous coreactant-free ECL sensor for UA is established. In this case, zeolitic imidazolate framework-8 (ZIF-8) assembles the carbon dots via π-π stacking and hydrogen bonding as an ECL emitter. Owing to the pore confinement effect, ZIF-8 works as an anchor to pre-concentrate abundant dissolved O2 and catalytic reduction of O2 by the pyridine nitrogen in the structure, generating massive reactive oxygen species (ROSs) to achieve an enhanced ECL intensity without the exogenous coreactant.
H2O2 in living cells can generate hydroxyl radical to damage proteins and tissues, which is a significant intracellular signaling molecule and disease marker. Thus, sensitive detection of H2O2 is of the utmost importance. Tian et al. measured the concentration of H2O2 by using the peroxidase-mimetic N-doped porous carbon-containing Fe (Fe/N-C) nanocomposites to fabricate a noble metal-free electrode. Fe/N-C nanocomposites are from the pyrolysis of NH2-MIL-101(Fe) MOFs, which significantly convert H2O2 to superoxide radical and oxidate luminol to enhance the ECL intensity. Besides, various noninvasive electrodes for the detection of H2O2 are achieved. For example, Jian et al. evaluated a solid-state ECL sensor for in situ H2O2 detection at the cellular level (Figure 3). In this work, TiO2 nanotubes (TiNTs) grow around a Ti wire and act as an ECL emitter, MIL-88B(Fe), as the "enzyme" decomposes H2O2 to superoxide radical, leading to an increasing ECL intensity. The high peroxidase-like activity of MIL-88B(Fe) toward H2O2 is attributed to the high dispersed Fe node in MIL-88B(Fe) structure. Moreover, owing to the photocatalytic activity of MIL-88B(Fe)@TiNTs, the sensors can be self-cleaned and reused. These merits of the ECL sensors pave the way for promising applications in the environment and clinic.
Antibiotic medicine has received a broad range of attention due to its antivirus and anti-inflammation. Overdose intake may pose potential risks to human health, so it is essential to know the exact dose. A sensitive and selective ECL aptasensor for kanamycin detection is constructed via the catalysis of Au@HKUST-1 towards the perylene derivative (PTC-Cys)/S2O82- system. Owing to the large surface area and the potential of binding with sulfhydryl, Au@HKUST-1 is used to fabricate the electrode. The fabricated electrode obtains an enhanced ECL intensity in comparison to the bare electrode, which is a result of the fact that Au@HKUST-1 accelerates the electrochemical reduction process of S2O82- to produce more sulfate radical anions (SO4-•), achieving an enhancement of ECL intensity. After the aptamer of kanamycin binds to the fabricated electrode, the ECL intensity is decreased. In the presence of kanamycin, aptamer on the electrode will be drawn away from the surface of the electrode, and the ECL intensity recovered. As a tendency in analytical chemistry, dual-target analysis is widely studied in ECL sensors. Feng et al. developed a dual gears ECL aptasensing strategy for kanamycin and neomycin detection using MIL-53(Fe) @CdS-PEI as the ECL emitter and metal nanoparticle (Au NPs or Pt NPs) as both ECL quencher and enhancer. The sensitive detection of multiple antibiotic medicines with a single ECL emitter overcomes the difficulty that the similar properties and structure of kanamycin and neomycin couldn't be distinguished directly. Rutin, a biologically active flavonoid glycoside, is a kind of therapeutic medicine that has demonstrated a broad range of physiological functions. Nie et al. constructed GSH-Au NCs@ZIF-8 nanocomposites by utilizing the coordination interaction of Zn2+ with the carboxyl group in glutathione (GSH) protected Au NCs (GSH-Au NCs) and the nitrogen atom of 2-methylimidazole linkers. Then, a signal-off ECL sensor is proposed for rutin detection based on the ECL quenching of rutin toward GSH-Au NCs@ZIF-8 nanocomposites/triethylamine system (Figure 4).
With the increasing awareness of food safety, monitoring pesticide residue has caused widespread concerns. Imidacloprid, a widely used neonicotinoid insecticide, is detected by an "on-off" ECL sensor in the combination of molecularly imprinted polymer with ultrafine mixed-valence Ce-MOF nanowires (Figure 5). The proposed ECL sensor exhibits a high selectivity by using molecular imprinting (MIP) technology, while the sensitivity is not satisfied. Acetamiprid, a type of neonicotinoid insecticide, is detected by an "off-on" ECL aptasensor based on the resonance energy transform from molybdenum disulfide quantum dots (MoS2 QDs) to a Zr-based metal-organic framework NH2-UiO-66. First, the aptamer of acetamiprid is immobilized on the MoS2 QDs fabricated electrodes. NH2-UiO-66 acts as the signal element to link with pDNA which then matches with the aptamer to obtain the "off" status of the aptasensor. With the introduction of acetamiprid, the base pair of pDNA to aptamer is interrupted and produces an "on" ECL signal. With a similar "off-on" design principle, omethoate, a kind of organophosphorus pesticide, is detected by using the MoTe2 nanoparticles doped ZIF-8 to electrocatalytically reduce the coreactant S2O82− to SO4-•. Meanwhile, a sensitive ECL sensor is constructed for malathion detection, another kind of organophosphorus pesticide. In this work, Fe-based metal-organic framework NH2-MIL-88(Fe) is worked as the ECL signal emitter carrier and signal enhancer for the CdTe QDs-S2O82- system. As the variety of pesticides increases, simultaneous detection of multiple pesticide residues is necessary for food safety. Accordingly, a dual-signal ECL sensor is proposed for simultaneous detection of acetamiprid and malathion by utilizing hollow Cu/Co-MOF-luminol and g-C3N4 as ECL emitters. The two signal probes are used in the preparation of ECL sensors for realizing two pesticides detection at the same time, and thus provide a new idea for simultaneous detection of multiple pesticides.
Nucleic Acid Sensing. As a powerful method, MOFs-based ECL sensors have gradually expanded from the sensitive and selective detection of small molecules to nucleic acid (eg. DNA and RNA). Since the abnormal expression of nucleic acid is an indicator of malignantly cellular physiological and pathological processes, plenty of strategies for sensitively and selectively nucleic acid quantification are developed, which mainly relied on the principle of specific base pairing between capture probe and target nucleic acid by using MOFs as ECL emitters or carriers to construct MOF-based ECL sensors. With the research going on, a series of signal amplification techniques are integrated into the MOF-based ECL sensors for nucleic acids detection, such as hybridization chain reaction (HCR), enzyme-assisted signal amplification, DNA walking machine, etc. By using lanthanum (La) MOFs as an ECL emitter, a MOFs-based ECL sensor for p53 gene detection is established on basis of a double-stranded DNA (dsDNA), triggering the co-quenching effect of crystal violet (CV) to LaMOFs. Target p53 gene works as a bridge for the electron transfer between excited LaMOFs and CV molecules. CV molecules that insert the p53 gene dsDNA grooves are used as an electron acceptor of LaMOFs to further quench the emission of LaMOFs. When the target p53 gene dsDNA is anchored on the LaMOFs modified electrode, the electron transfer between excited LaMOFs and CV molecules is triggered, resulting in a decreased ECL intensity. The breast cancer 1 gene (BRCA1) is a reliable biomarker in early cancer and mutation. Wang et al. present a coreactant-free ECL biosensor for sensitive BRCA1 detection with a flower-like metal-organic framework microsphere (ZnMOF(Ru)) as an ECL emitter. The prepared ZnMOF(Ru) displayed a 53-fold ECL intensity to that of Ru(bpy)32+. Such work would encourage more inspiration to develop novel MOFs in ECL sensors.
MicroRNAs (miRNAs), a diminutive noncoding RNA, are related to cancers. Numerous ECL sensors have been constructed for the selective and sensitive detection of miRNAs. For example, a Faraday-cage ECL biosensor for the sensitive detection of miRNA-141 is reported. In this work, the prepared Ru-MOFs nanosheets are not only used as an ECL emitter but also extend the Outer Helmholtz Plane of the magnetic electrode, thus improving the sensitivity via Faraday cage-type strategy. Signal amplification techniques, as a powerful way to improve sensitivity, have been widely used in ECL biosensors for the detection of miRNA. In combination with the exonuclease III (Exo III)-aided target recycling amplification strategy, Yang et al. take the conductively bimetallic MOFs (NiCo-HHTP) nanorods as an ECL emitter to construct an ultrasensitive ECL biosensor for miRNA-141 detection. MOFs-based ECL sensor integrated with signal amplification strategies is also used for miRNA-21 detection. Because miRNA-21 is a kind of cancer-related microRNA, sensitive detection of miRNA-21 is of prime importance. Jiang et al. designed an ECL biosensor for miRNA-21 detection via a hybrid Co-MOF-ABEI/Ti3C2Tx composite as an ECL emitter and a duplex-specific nuclease (DSN)-assisted signal amplification strategy. Meanwhile, an "off-on" ECL biosensor for miRNA-21 detection is developed by using zinc-based perylene-3, 4, 9, 10-tetracarboxylate MOFs (Zn-PTC MOFs) as a new ECL emitter as well as the exonuclease III-stimulated target cycling and Zn2+-dependent DNAzyme-assisted cycling dual amplification strategies (Figure 6). With the appearance of target miRNA-21, the exonuclease III-stimulated target cycling is triggered to produce a simulated target. Then, the simulated target hybrids with the ferrocene (Fc)-labeled capture probe (Fc-S1) that assembles on the Zn-PTC MOFs modified electrode. At last, with the aid of cofactor Zn2+, the DNAzyme starts to cleave Fc-S1, resulting in Fc being far away from the electrode surface. In this principle, the prepared Zn-PTC MOFs eliminate the aggregation-induced quenching of PTC monomers because the MOFs structure amplified the distance of PTC perylene rings. This method proposed a convenient strategy to enhance ECL by eliminating the aggregation-induced quenching effect of polycyclic aromatic hydrocarbons (PAHs) via coordinative immobilization of PAHs within MOFs and opening a new path for MOFs application in ECL sensors.
MOFs-based ECL sensor integrated with signal amplification strategies is also used for other miRNA detection. For example, a dual potential ratiometric ECL biosensor is proposed for miRNA-133a detection with HCR amplification reaction and Zn-MOFs. The prepared Zn-MOFs are utilized as a single ECL emitter while it exhibits both cathode and anode emissions. With the increase of miRNA-133a concentration, the amount of N, N-diethylethylenediamine (DEAEA) which is introduced by HCR amplification reaction gradually increases, leading to the ECL intensity of the anode being increased and the cathode being decreased. Further, the ECL intensity ratio of the anode to cathode is extremely increased. This study extends the perspective for the design of novel MOFs in the application of ratiometric ECL biosensors.
Protein Sensing. MOFs-based ECL sensors have been widely applied in the realm of protein detection as the sensitive detection of the overexpressed protein is quite important for early clinical diagnosis. According to the principle of specific recognition between aptamers and target protein or specific immunoreactions between antibody and antigen as well as an ECL reporter that enhances or quenches ECL signal, a series of MOFs-based ECL sensors is established. For instance, procalcitonin (PCT) with a 116-amino-acid protein which is a biomarker of septicemia is detected by a near-infrared ECL biosensor. The biosensor is fabricated by a combination of the antenna effect of ECL emitter Eu-MOFs and the efficient catalysis of the coreaction accelerators CoS2 hollow nanoboxes. Meanwhile, a dual-quenching sandwich-type ECL biosensor for PCT detection is proposed based on the ECL-RET from isoreticular metal-organic framework-3 composite (NGQDs@IRMOF-3@N-GQDs) to Au nanoparticles modified zinc oxide nanorod (ZnO@Au). Also, a similar dual-quenching strategy is used to realize PCT detection where the catechol or benzoquinone units of PDA structure and Cu2+ in Fe3O4@PDA-CuxO-Ab2 quenching the ECL emission of Ru(bpy)32+ doped hollow MIL-101(Al)-NH2 (Ru@MIL-101). Insulin plays a crucial role in the regulation of carbohydrate and fat metabolism and can be medically used for diabetes and related diseases. Ma et al. measured insulin sensitively through a sandwich-type ECL biosensor by energy transfer from the ECL emitter Au nanoparticle-doped Pb(II)-β-cyclodextrin MOFs (Au@Pb-β-CD) to chitosan/Ru(bpy)32+/silica nanoparticles (CRuSi NPs). In another work, Ru(bpy)32+ encapsulated UiO-67 (UiO-67/Ru(bpy)32+) nanocomposite and gold nanoparticles doped silica (Au@SiO2) are used as the ECL−RET donor and acceptor, respectively, for insulin detection.
Cardiac troponin I (cTnI) is considered as an important biomarker of acute myocardial infarction. In consequence, 2D Ru-MOFs nanosheet (Ru-MOFs NSs) is prepared for a "signal-on" ECL detection of cTnI by using tris(4, 4′-dicarboxylic acid-2, 2′-bipyridyl) ruthenium(II) (Ru(dcbpy)32+) as an organic ligand to coordinate with zinc metal, and polyvinylpyrrolidone (PVP) as a structure-directing agent to promote the formation of MOFs nanosheet (Figure 7). The highly exposed Ru(dcbpy)32+ reactive sites are beneficial to enhancing the ECL intensity and improving the sensitivity of the biosensor. Except for the 2D MOFs, octahedral Co2+-based ZIF-67 MOFs decorated with luminolcapped Ag nanoparticles (luminol-AgNPs@ZIF-67) exhibit an outstanding ECL performance in aqueous solution (about 115-fold enhancement beyond the traditional luminol system). The application in a label-free ECL biosensor realizes the ultrasensitive detection of cTnI. In addition, because the large surface can load massive ECL emitter, rod-like amino-functional MOFs (NH2-MIL(Fe)) are decorated with carbon nitride nanosheet (CNNS) to form CNNS@NH2-MIL(Fe). The prepared CNNS@NH2-MIL(Fe) works as an effective ECL emitter and the aminofunctionalized titanium carbide (N-Ti3C2) nanosheet acts as an excellent substrate in constructing a highly sensitive ECL biosensor. This biosensor displays a satisfactory sensitivity for the detection of cTnI.
Thrombin (TB) is a specific serine protease and is regarded as a biomarker in many blood-related diseases, such as hemostasis, cardiovascular disease, and thrombosis.[88, 89] Hence, its sensitive detection is significantly important for disease diagnosis. Fang et al. report a label-free ECL aptasensor for TB determining. In this work, aptamer, a single nucleic acid that can specifically recognize and bind to the target, is utilized to recognize and bind TB. Zinc proto-porphyrin IX grafted ZIF-8 (ZnP-NH-ZIF-8) is synthesized as an enhanced ECL emitter, and its high ECL efficiency is contributed by the efficient oxygen reduction reaction activity. The ECL intensity of ZnP-NH-ZIF-8 modified electrode is decreased with the increase of TB concentration because the steric hindrance is produced by TB which is recognized and bound with aptamers. With similar strategy, a porphyrin Zr-MOFs (PCN-222) is applied in the ECL aptasensor for TB detection. Due to the good conductivity, abundant porosity, and coreactant acceleration of ECL emitter PCN-222, the electrode fabricated with PCN-222 displays an excellent performance. This method provides a simple strategy for the synthesis of PCN-222 with an excellent ECL performance but the stability is susceptible. Combined with the RecJf exonuclease assistant target recycling amplification technology, the precise detection of TB is realized by using a ZIF-8 nanocomposite (Ru-PEI-L-lys-ZIF-8). In the meantime, the enzyme-free amplification technology of catalytic hairpin assembly (CHA) and a hollow Zr6-based MOFs nanocomposite (HHRu-UiO-66-NH2) is applied in measuring TB. The hierarchical-pore shell and hollow cavity of hollow hierarchical UiO-66-NH2 provide large space for immobilizing plentiful Ru(bpy)2(mcpbpy)2+ (mcpbpy = 4-(4′-methyl-[2, 2′-bipyridin]-4-yl) butanoic acid) and permit the electron and coreactant diffusion, further promoting the ECL efficiency.
Trypsin is a vital diagnostic indicator for renal failure and pancreatic cancer, and it is eagerly anticipated to detect it efficiently. Accordingly, an "on-off" peptide-based biosensor is proposed by using a copper-based MOFs nanocomposite (JUC-1000-Fe3O4@Au) and core-shell Ag@CeO2 NPs. JUC-1000-Fe3O4@Au nanocomposite assembles a heptapeptide (HWRGWVC, HGC) that can be specifically recognized and cleaved by trypsin to develop a simple and reliable trypsin identification system (JUC-1000-Fe3O4@Au-HGC). Ag@CeO2 NPs modified GCE (Ag@CeO2 NPs/GCE) is used as the sensing interface to capture the JUC-1000-Fe3O4@Au-HGC. Then, with the introduction of trypsin, HGC is cleaved by trypsin, leading to JUC-1000-Fe3O4@Au being released from the sensing surface, and the ECL intensity is decreased. This work provides a primary reference for the monitoring of trypsin.
Prostate-specific antigen (PSA) is the most sensitive biomarker for the detection of prostate and breast cancer. A labelfree ECL biosensor is designed by using a silver nanoparticles-doped Pb(II) metal-organic framework (Ag@Pb(II)-β-CD) as substrate material to modify GCE and form a sensing platform. Moreover, Fe(II)-MOFs/Au/G-quadruplex are utilized as both ECL quencher and enhancer to develop a target-triggered ratiometric ECL biosensor for the quantification of PSA based on the competition of target PSA and a DNA linker. When introducing PSA, the competition between PSA and DNA linker is triggered, resulting in the ECL intensity of the luminol-H2O2 system being decreased and the QDs recovered. As a result, the accurate detection of PSA is realized. With the amplification of artificial mimic enzyme NH2-MIL(53)-Fe nanocomposite (C3N4/NMF) toward luminol-H2O2 system and the enrichment of magnetic nanoparticles, a sandwich-type ECL biosensor is rationally designed for PSA detection.
Carcinoembryonic antigen (CEA) is a glycosylated protein and is considered as an available indicator for liver and colorectal cancer. Accurate evaluation of CEA is significant and necessary for understanding cancer progression. Therefore, a ZIF-8@GO composite derived nanomaterial (AuNP@NPCGO) acts as a substrate for immobilizing CEA antibody and Ru(phen)32+-doped silica nanoparticles act as ECL emitter to fabricate a sandwichtype ECL biosensor for CEA detection. In another work, CdSe QDs decorated MIL-101 (MIL-101-CdSe) composite was synthesized by hydrothermal method for sensitive detection of CEA. With increasing the concentration of CEA, the steric hindrance of electron transfer and K2S2O8 to MIL-101-CdSe is increased, leading to a decreasing ECL intensity. By utilizing the similar above-mentioned design basis, both Cu2+ doped terbium luminescent metal-organic framework (Cu: Tb-MOF) and H2O2 exploit as a co-reaction accelerator to develop a label-free ECL biosensor for progastrin-releasing peptide (ProGRP, a vital biomarker of small-cell lung cancer) detection.
Amyloid-β protein (Aβ) is a 39-43 amino acid residue that serves as a predictive biomarker for Alzheimer's disease (AD). Consequently, it is of great significance to provide an effective strategy for Aβ qualification. Wang et al. proposed an ECL biosensor for Aβ in which Fe3O4@PPy-Au nanoparticles are adopted to fabricate the sensing platform and the Co-MOFs/ABEI is not only adopted as an ECL emitter but also a co-reaction promoter to generate more ROSs for the amplification of ECL response. At the same time, an effective dual-quenching ECL tactic is established based on the ECL-RET behavior between bimetallic NiFe-based nanocube MOFs nanocomposite (Au@NiFe MOFs) and three-dimensional Ru(bpy)32+/zinc oxalate MOFs. Except for energy transfer, the electron transfer also causes the quenching effect. In virtue of this fact, Zr-MOFs are used as an ECL emitter to fabricate the sensing platform for epinephrine (EP) detection.
Protein kinase A (PKA) is one kind of protein kinase that catalyzes peptide phosphorylation in the regulation of the metabolism and has been regarded as an indicator for disease diagnosis.[105, 106] Zhang et al. achieved sensitive detection of PKA using zirconium-(zinc)porphyrin MOFs (MOF-525-Zn) as a multiplex functional platform. First, Zr-O clusters of MOF-525-Zn serve as the recognition sites to recognize and bond with the PKA-catalyzed peptide phosphorylation through phosphate groups. Then, zinc tetrakis(carboxyphenyl)-porphyrin (ZnTCPP) in MOF-525-Zn reacts with O2 enriched by the MOFs porous structure to generate 1O2. Finally, ZnTCPP also works as an ECL emitter to produce ECL emissions under the 1O2 and the presence of tetraoctylammonium bromide (TOAB). Hence, a multiplex functional platform for the detection of PKA is realized. The platform possesses oxygen nanocage, electron media, and a binding site in the analysis system which would have potential applications in accurate clinic diagnostics.
Mucin-1 (MUC-1) is a sort of transmembrane glycoprotein that is widely recognized as a predominant tumor marker for prostate cancer, implicating the importance of sensitive detection of MUC-1 for early evaluation of prostate cancer. An ABEI functionalized Fe-based MOF (ABEI/MIL-101(Fe) with intrinsic mimic peroxidase activity and bimetallic AgPt hollow nanospheres (AgPt HNSs) with a large surface area is utilized for MUC-1 detection. In another report, on the basis of the quenching effect of thionine on the zinc meso-tetra(4-sulfonatophenyl) porphine (Zn-TP) linked Zn-based MOFs (Zn-TP@Zn-Bp-MOFs), a label-free ECL biosensor is constructed. It is worth noting that the ECL efficiency of Zn-TP@Zn-Bp-MOFs is greatly enhanced due to the synergistic effects of high electrocatalysis and more excited electrons harvested. Amplification technology is integrated into the ECL biosensors for MUC-1 detection to improve sensitivity. For instance, a mesoporous Ru-functionalized Zr-based MOF (Ru-PCN-777) is used to fabricate a sensitive ECL biosensor that combines the proximity-induced intramolecular DNA strand displacement and nicking endonuclease Nt.BbvCI assisted DNA recycling amplification. Ru-PCN-777 provides a large effective site for the sensing platform to capture the abundant DNA probe and is used as the ECL emitter carrier to prevent the leakage of Ru. Apart from a single amplification, multiple amplification technologies are also used in MOFs-based ECL biosensors for MUC-1. Yao et al. use a Ru doped 2D MOFs nanoplate (Ru@Zr12-BPDC) as the ECL emitter to establish an ultrasensitive ECL biosensor that is based on the double amplification strategies of Exo III-assisted recycling and CHA. Likewise, a new ECL emitter (Hf-based MOFs (Hf-TCBPE)) is discovered and applied in an "off-on" ECL biosensor by twice using the Exo III-assisted recycling amplification. Hf-TCBPE exhibits an outstanding ECL performance primarily because the rigid MOFs structure restricts the intramolecular rotation of phenyl rings and loads plentiful tetraphenylethylene.
Alpha-fetoprotein (AFP) is an oncofetal glycoprotein and is treated as a hepatocellular carcinoma indicator. CdSnS NCs functioned magnetic MOFs (MMOF@CdSnS) nanoparticles are applied in AFP detection. MMOF@CdSnS nanoparticles behave as both the ECL emitter and the AFP antibody carrier, which specifically recognize and bind with AFP to form MMOF@CdSnS@Ab@Ag. Magnetic ITO electrode is fabricated with Ag NPs modified reduced the graphene oxide (Ag NPs@rGO) nanocomposite with high conductivity and magnetically enriches the MMOF@CdSnS@Ab@Ag. With this principle, AFP is detected with the merits of easy separation and enriched efficiency. Alternatively, ultrathin Co/Ni-based MOFs nanosheets are used for AFP detection. The use of Co/Ni-based MOFs nanosheets improves the ECL performance of the luminol-AgNPs system.
Bacteria, and Virus Sensing. Vibrio parahaemolyticus (VP) is a bacterial pathogen, food pollution with VP may lead to food poison. Thus, the rapid and reliable detection of VP is crucial for protecting the health of human beings. Wei et al. construct a Faradic cage-type aptasensor for dual-mode detection of VP. The dual-mode detection is performed by using a VP recognition aptamer Apt2 labeled two-dimensional Pb2+-Ru-MOF as the signal unit (Pb2+-Ru-MOF@Apt2), in which Ru produces the ECL signal and Pb2+ can be reduced to produce the electrochemical signal. When VP exists, the aptamer1 on the electrode selectively captures VP, and then Pb2+-Ru-MOF@Apt2 specific recognizes VP and binds to it, so that the two-dimensional signal unit is close to the electrode to form a Faraday cage-type aptasensor. The use of two-dimensional MOF provides good conductivity and a channel for electron transfer by forming the Faraday cage. This method can also be extended for the detection of other large-volume biological targets.
As a member of the Flaviviridae family virus, Zika virus (ZIKV) with a single-stranded RNA has formidable neurotropic toxicity and teratogenicity. Zhang et al. use a g-C3N4 and Au NPs functioned Zr-based metal-organic gel (Au NPs & g-C3N4@Zr-MOG) as electrode matrix and Fe-MIL-88 MOFs as nanotag to measure the ZIKV concentration. It should be noted that the Fe-MIL-88 MOFs own double quenching effect for the ECL reaction because they not only behave as ECL acceptors of Au NPs & g-C3N4@Zr-MOG in ECL-RET but also consume the coreactant. Porcine epidemic diarrhea virus (PEDV) is a fast infectious coronavirus that causes serious diarrhea and dehydration. Ma et al. proposed an ultrastable ECL biosensor for PEDV detection based on the ECL emitter porphyrin Zr-based MOFs (PCN-224) and the accelerating function of TiO2 nanoparticles.
Escherichia coli (E. coli) causes urinary tract infections and arthritis in certain conditions, which has aroused significant interest involving its sensitive detection. Recently, a variety of ECL biosensors are constructed for the sensitive detection of E. coli. Sun et al. proposed an ECL biosensor for E. coli BL21 detection based on a Ru-Con A coated NH2-MIL-53(Al) as the sensing interface (Figure 8). Ru-Con A is formed by a ruthenium complex tagged Con A. Attaching Ru-Con A to the NH2-MIL-53(Al) surface remarkably enhanced the ECL signal, which conduced to the sensitivity of detection. The proposed biosensor simultaneously assessed the antibacterial susceptibility of several antibiotics to normal and drug-resistant E. coli strains.
CONCLUSION AND PERSPECTIVES
By virtue of the distinct properties of large surface area, tunable porosity and structure, high molecule loading capacity, versatile functionality, and improved biocompatibility, MOFs are found to be a promising candidate and employed in the ECL sensors for diverse analytes. This summary presents the recent advances in MOFs-based ECL sensors for various target detection with representative examples. The target covers the metal ions, small molecules, nucleic acids, proteins, bacteria, and viruses in the realm of the hotspot. In this regard, MOFs are utilized as ECL emitter, carrier of ECL emitter, coreactant accelerator, material for the construction of sensing interfaces, an active site in the mimic enzyme, ECL quenching material, and the acceptor or donor in the ECL-RET system. The relationship between ECL performance and the characters of MOFs is also illustrated. First, the large surface area of MOFs is beneficial for loading massive functional materials to fabricate sensing platforms or loading plentiful ECL emitters to improve the sensitivity of ECL sensors. Second, due to the high porosity and adjustable structure of MOFs, the active sites are accessible and the diffusion of the electrons and energies is rapid by the channels, further improving the efficiency of ECL reaction and enhancing the ECL intensity. Third, the metal nodes of MOFs exhibit excellent catalytic activity and synergic effect in multiple components for signal transduction in MOFs-based ECL sensors which also contribute to the ECL reaction efficiency and ECL intensity. Although great achievements are obtained, the MOFs-based ECL sensors are actually in their infancy. Many challenges and opportunities of MOFs are faced in ECL sensor applications. To further facilitate the ECL sensing application and improve the performance in ECL sensors, the following aspects deserved to be carefully considered in the future: (1) To avoid the erroneous response triggered by randomness, MOFs with two or multiple emission centers should be further exploited to construct ratiometric MOFs-based sensors for accurate target detection. (2) To find more aqueous stable and low toxic MOFs for ECL sensors, theoretical calculations should be incorporated into the design process of MOFs to decrease the cost of practice studies. (3) More MOFs with different ECL spectra or potentials should be synthesized for simultaneous multiple target detection by integrating the functional materials or groups on MOFs. (4) For achieving high sensitivity of MOFs-based ECL sensors, more amplification strategies should be considered. (5) The current developed MOFs-based ECL sensors are in the laboratory stage and lack of practical applications. Therefore, to minimize the gap between laboratory study and practical application, portable electrode or instrument should be taken into account. For instance, MOFs are used to fabricate the paper-based electrode, ITO electrode, printing electrode, or wearable flexible electrode, and then record and analyze the signal with a workstation similar to the PalmSens USB workstation or phone to realize the practical detection of the target. In a word, MOFs-based ECL sensors have made significant progress in research, while further study is still needed to establish sensitive, accurate, rapid, and cost-effective sensors and expand the potential applications in more extensive and broader fields. We insist a productive future for MOFs-based ECL sensors can be expected with the collision and strengthen cooperation between different subjects.
ACKNOWLEDGEMENTS: Financial support from the Foundation of Suzhou University of Science and Technology (No. 332114415), the National Natural Science Foundation of China (21904092), and Jiangsu Laboratory for Biochemical Sensing and Biochip is acknowledged. COMPETING INTERESTS
The authors declare no competing interests.
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Figure 1 The mechanism for Hg2+-responsive disassembly of Ru-MOFs and release of the guest material of Ru(bpy)32+. Reproduced with permission from Ref.
Figure 2 The schematic diagram of Au@Pb-β-CD applied in the ECL sensor for the detection of Cr6+. Reproduced with permission from Ref.
Figure 3 Schematic representation of the preparation of (A) MIL-88B(Fe)@TiNTs/Ti; (B) Formation mechanism of MIL-88B(Fe)@TiNTs/Ti; (C) Schematic illustration of the solid-state ECL sensor detecting H2O2 in living cells. Reproduced with permission from Ref.
Figure 4 (A) Schematic diagram for the preparation of GSH-Au NCs@ZIF-8 and the ECL-enhanced mechanism; (B) Schematic illustration of ECL detecting rutin based on GSH-Au NCs@ZIF-8/TEA system. Reproduced with permission from Ref.
Figure 5 (a) Biomimetic construction of the UMV-Ce-MOF nanowires and (b) Preparation of an imidacloprid MIECS based on the UMV-Ce-MOF nanowires. Reproduced with permission from Ref.
Figure 6 (A) Preparation of Zn-PTC. (B) Exo III-stimulated target cycling process. (C) The application of Ru@MIL-101 in PCT detection. Reproduced with permission from Ref.
Figure 7 The schematic diagram for the fabrication process of ECL immunesensor. Reproduced with permission from Ref.
Figure 8 (A) Schematic illustration for the ECL biosensor for detection of E. coli BL21 and (B) antimicrobial susceptibility assays of representative antibiotics against E. coli BL21 and NDM-1 E. coli BL21. Reproduced with permission from Ref.
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