A Chemiluminescence Sensor for the Detection of α-Fetoprotein and Carcinoembryonic Antigen Based on Dual-Aptamer Functionalized Magnetic Silicon Composite.

In this work, a dual-aptamer functionalized magnetic silicon composite was prepared and used to construct a chemiluminescence (CL) sensor for the detection of α-fetoprotein (AFP) and carcinoembryonic antigen (CEA). First, SiO2@Fe3O4 was prepared, and polydiallyl dimethylammonium chloride (PDDA) and AuNPs were sequentially loaded on SiO2@Fe3O4. Subsequently, the complementary strand of CEA aptamer (cDNA2) and the aptamer of AFP (Apt1) were attached to AuNPs/PDDA-SiO2@Fe3O4. Then, the aptamer of CEA (Apt2) and G quadruplex peroxide-mimicking enzyme (G-DNAzyme) were sequentially connected to cDNA2, leading to the final composite. Then, the composite was used to construct a CL sensor. When AFP is present, it will combine with Apt1 on the composite to hinder the catalytic ability of AuNPs to luminol-H2O2, achieving AFP detection. When CEA is present, it will recognize and bind with Apt2, so G-DNAzyme is released to solution and catalyzes the reaction of luminol-H2O2 to achieve CEA determination. After the application of the prepared composite, AFP and CEA were detected in the magnetic medium and supernatant, respectively, after simple magnetic separation. Therefore, the detection of multiple liver cancer markers is realized through the CL technology without additional instruments or technology, which broadens the application range of CL technology. The sensor for detecting AFP and CEA shows wide linear ranges of 1.0 × 10-4 to 1.0 ng·mL-1 and 0.0001-0.5 ng·mL-1 and low detection limits of 6.7 × 10-5 ng·mL-1 and 3.2 × 10-5 ng·mL-1, respectively. Finally, the sensor was successfully used to detect CEA and AFP in serum samples and provides great potential for detection of multiple liver cancer markers in early clinical diagnosis.

Ultrasensitive Electrochemiluminescence Biosensor with Silver Nanoclusters as a Novel Signal Probe and α-Fe2O3-Pt as an Efficient Co-reaction Accelerator for Procalcitonin Immunoassay.

Herein, a high-efficiency biosensor based on ternary electrochemiluminescence (ECL) system was constructed for procalcitonin (PCT) detection. Specifically, silver nanoclusters (Ag NCs) with stable luminescence properties were prepared with small-molecule lipoic acid (LA) as the ligand, and its ECL emission in persulfate (S2O82-) was first reported. Meanwhile, the prepared Ag NCs possessed ligand-to-metal charge-transfer characteristics, thus transferring energy from LA to Ag+ for luminescence. Based on the small particle size, good biocompatibility, and molecular binding ability, Ag NCs-LA was used as an ideal luminescent probe. In addition, α-Fe2O3-Pt was introduced to facilitate the activation of S2O82-, thereby generating more sulfate radicals to react with the free radicals of Ag NCs to enhance ECL emission. The synergistic effect of the variable valence state of transition metals and high catalytic activity of noble metals endows α-Fe2O3-Pt with excellent catalytic ability for S2O82-. Importantly, the sensing mechanism was systematically demonstrated by UV-vis, fluorescence, and ECL analysis, as well as density functional theory calculations. At last, NKFRGKYKC was designed for specific immobilization of antibodies, thus releasing the antigen binding sites to improve the antigen recognition efficiency. Based on this, the developed biosensor showed high sensitivity for PCT detection, with a wide linear range (10 fg/mL-100 ng/mL) and a low detection limit (3.56 fg/mL), which could be extended to clinical detection of multiple biomarkers.

A novel flower-shaped Ag@ZIF-67 chemiluminescence sensor for sensitive detection of CEA.

In this work, a chemiluminescence (CL) aptasensor for sensitive carcinoembryonic antigen (CEA) detection was constructed based on the CL system of luminol-H2O2-NaOH. Magnetic carbon nanotubes (MCNTs), as the base material, was modified with CEA-aptamer and DNA1, and was combined with the novel flower-shaped Ag@ZIF-67 of modified with DNA2 through the principle of base complementary pairing. CEA combined with aptamer when it existed in the solution. At the same time, MCNTs was adsorbed at the bottom of the container under the influence of external magnetic field, and Ag@ZIF-67 enhanced the CL signal. The CL aptasensor demonstrated high selectivity and sensitivity for CEA in human serum sample with (1-4): a detection limit of 4.53 × 10-3 ng/mL in case the detection range was 0.05-500 ng/mL. Furthermore, the proposed method had been shown great potential in cancer diagnosis.

Zinc-Based Metal-Organic Framework with MLCT Properties as an Efficient Electrochemiluminescence Probe for Trace Detection of Trenbolone.

In this work, we utilized polycyclic aromatic hydrocarbon (PAH) derivatives as ligands to develop a zinc-based metal-organic framework (Zn-MOF) as an effective detection probe to construct an electrochemiluminescence (ECL) sensor for trenbolone detection. As traditional ECL emitters, PAHs and their derivatives have limited luminescence efficiency because of the aggregation-induced quenching (ACQ) effect. Therefore, Zn-PTC was designed by the coordination of 3,4,9,10-perylenetetracarboxylic (PTC) in the MOF to eliminate the ACQ effect. Meanwhile, Zn-PTC formed based on an aromatic ligand possessed the metal-to-ligand charge-transfer (MLCT) effect, which could transfer the energy of Zn2+ to the aromatic ligand for strong luminescence. The ECL efficiency of Zn-PTC was calculated to be approximately 2.2 times that of the ligand (K4PTC). Second, the Ag@Fe core-shell bimetallic nanocrystal was prepared for efficient activation of persulfate (S2O82-), thereby generating more sulfate radicals (SO4•-) to further promote ECL emission. According to ECL characterizations, UV-vis and fluorescence spectra, and density functional theory calculations, the luminescence and signal amplification mechanisms were investigated. In addition, NKFRGKYKC (NKF) was introduced as an affinity ligand to directionally immobilize the target antibodies, thus releasing specific sites in their Fab fragment to enhance binding activity. Based on the above strategies, the constructed biosensor exhibited high sensitivity, realizing trace detection of TBE with a wide detection range (10 fg/mL-100 ng/mL) and a low detection limit (3.28 fg/mL). This study provided an important reference for sensitive monitoring of steroid pollutants in the environment.

A Portable Microfluidic-Based Electrochemiluminescence Sensor for Trace Detection of Trenbolone in Natural Water.

In this study, a portable electrochemiluminescence sensor chip was designed for trenbolone (TBE) trace detection in environmental water. First, a stable ECL signal was obtained with low-toxicity 3,4,9,10-perylenetetracarboxylic acid (PTCA) as a luminophore and persulfate (S2O82-) as a coreactant. Second, hollow-structured Cu2MoS4 was introduced as a coreaction accelerator to catalyze S2O82- reduction. The reversible conversion of the mixed-valence transition metal ions in Cu2MoS4 (Cu+/Cu2+ and Mo4+/Mo6+) greatly promoted the generation of the sulfate radical (SO4•-). Meanwhile, the special porous structure of Cu2MoS4 possessed a large specific surface area, thus enhancing its catalytic performance. Based on these enhancement mechanisms, a strong ECL signal was acquired, which improved the detection sensitivity of the constructed sensor. Importantly, a microfluidic chip was introduced for sensing detection, thereby improving the practicality of the sensor. The developed sensor chip was miniature and portable, exhibiting high sensitivity for TBE detection with a wide linear range (10 fg/mL-100 ng/mL) and lower detection limit (3.32 fg/mL). This was of great significance for timely and rapid analysis of steroid pollutants in natural water.

Highly Efficient PTCA/Co3O4/CuO/S2O82- Ternary Electrochemiluminescence System Combined with a Portable Chip for Bioanalysis.

Herein, we reported an efficient electrochemiluminescence (ECL) biosensor chip for sensitive detection of neuron-specific enolase (NSE). First, 3,4,9,10-perylenetetracarboxylic acid with good luminescence characteristics was used as a luminophore to obtain a stable ECL signal. Subsequently, hollow porous Co3O4/CuO concave polyhedron nanocages (CPNCs) were designed as co-reaction promoters to amplify the luminescence signals for highly sensitive trace detection of NSE. In brief, the rapid cyclic conversion of Co3+/Co2+ and Cu2+/Cu+ redox pairs could continuously catalyze the reduction of persulfate (S2O82-), thus providing a large number of essential active intermediates (SO4•-) for ECL emission. Meanwhile, the unique structure of Co3O4/CuO CPNCs possessed a large specific surface area, which greatly improved its catalytic efficiency. Third, NKFRGKYKC was developed as an affinity ligand for specific antibody fixation, which improved incubation efficiency and protected bioactivity of antibodies. Finally, we independently designed a microchip and applied it for ECL detection to improve the practical application ability of the sensor. The developed biosensor exhibited good sensitivity with a wide linear range (10 fg/mL to 100 ng/mL) and a low detection limit (3.42 fg/mL), which played an active role in the clinical application of sensing analysis.

Efficient ABEI-Dissolved O2-Ce(III, IV)-MOF Ternary Electrochemiluminescent System Combined with Self-Assembled Microfluidic Chips for Bioanalysis.

A signal-amplified electrochemiluminescent (ECL) sensor chip was developed for sensitive analysis of procalcitonin (PCT). Herein, we first prepared a self-enhanced luminophore, which enhanced ECL responses through intramolecular reactions. Second, Au-Pd bimetallic nanocrystals and mixed-valence Ce-based metal-organic frameworks (MOFs) were introduced as co-reaction promoters to facilitate the reduction of dissolved O2. Based on the synergistic catalysis of Au and Pd, the spontaneous cyclic reaction of Ce(III)/Ce(IV), and the high electrochemical active surface area of Ce(III, IV) MOF, a large number of superoxide anion radicals (O2•-) and hydroxyl radicals (OH•) were produced. Therefore, the luminescence efficiency of N-(aminobutyl)-N-(ethylisoluminol)-dissolved O2 (ABEI-O2) systems were greatly improved, providing a new prospect for the application of dissolved O2 in ECL analysis. In addition, the affinity peptide ligands were used for the directional connection of antibodies to provide protection for the bioactivity of the proposed sensor. Finally, the microfluidic technology was applied to ECL analysis to integrate the three-electrode detection system into the self-assembled microfluidic chip, which realized the automation and portability of the detection process. The developed sensor showed high sensitivity for PCT detection with a detection limit of 3.46 fg/mL, which possessed positive significance for the clinical diagnosis of sepsis.

A chemiluminescence aptasensor for sensitive detection of alpha-fetoprotein based on hemin@ZIF-67.

In this work, hemin@ZIF-67 composites were prepared and were used to construct a chemiluminescence (CL) aptasensor for alpha-fetoprotein (AFP) detection. Hemin is a catalytic porphyrin with two carboxylate groups that can covalently bond to metal ions. A hemin/ZIF-67 composite was prepared via covalent bonding between the carboxyl groups of hemin and the cobalt ion of ZIF-67, and these materials were characterized by scanning electron microscopy (SEM), infrared spectroscopy (IR), and X-ray diffraction (XRD). Hemin@ZIF-67 was used as the peroxidase material, and the aptamer of alpha-fetoprotein was modified on its surface by electrostatic adsorption. Then a simple CL aptasensor was constructed based on the CL system of luminol-H2O2-NaOH. Under the optimal conditions, the CL intensity value was linearly proportional to the concentration of AFP in the range of 4 × 10-10 to 200 × 10-10 mg/mL. The detection limit was 1.3 × 10-10 mg/mL. Thus the aptasensor enables highly sensitive and selective detection of AFP.

Chemiluminescence sensing of adenosine using DNA cross-linked hydrogel-capped magnetic mesoporous silica nanoparticles.

At present, a host of high-sensitivity and selective tumor marker detection methods play a central role in various research fields and assay platforms. Here, a method for DNA cross-linked hydrogel-capped magnetic core-shell structure mesoporous silica nanoparticles (Fe3O4@nSiO2@mSiO2) based on target stimulus was proposed. Specifically, Fe3O4@nSiO2@mSiO2 nanoparticles with nucleic acid promoter units were prepared. The promoter induces the predesigned modified polyacrylamide DNA strand to carry out hybridization chain reaction on the surface of Fe3O4@nSiO2@mSiO2 nanoparticles, forming a hydrogel coating on the surface of Fe3O4@nSiO2@mSiO2 nanoparticles. Under the stimulation of adenosine, Fe3O4@nSiO2@mSiO2 released the signal molecule luminol. Simultaneously, the MIL-101（Fe）material, a signal amplification molecule, was released from the DNA hydrogel. Compared with the traditional gated mesoporous silica system, the DNA hydrogel-coated Fe3O4@nSiO2@mSiO2 is not easy to leak and has a higher loading capacity. In addition, the optical background of DNA hydrogels is low, combined with MOFs materials, even about 1.4 × 10-10 M adenosine can be detected in this biosensor. Based on the combination of DNA hydrogels, MOFs conjugates and gating systems, the construction of biosensors will be more eye-catching, which will expand new ideas for biosensor platform manufacturing.

Peptide-Based Electrochemiluminescence Biosensors Using Silver Nanoclusters as Signal Probes and Pd-Cu2O Hybrid Nanoconcaves as Coreactant Promoters for Immunoassays.

Metal nanoclusters (NCs) possess high light stability and biocompatibility because of their unique quantum size effect, which has gradually become a new type of electrochemiluminescence (ECL) nanomaterial for immunoassays. However, the luminescence efficiency of metal NCs is too low to meet the needs of trace analysis, which limits its application. Herein, Ag NCs served as signal probes and Pd-Cu2O hybrid nanoconcaves served as coreaction promoters, developing a highly efficient peptide-based biosensor for neuron-specific enolase (NSE) detection. Utilizing the reversible cycle of Cu+/Cu2+ and the reduction characteristics of Pd NPs, Pd-Cu2O greatly accelerates the reduction of S2O82-. Meanwhile, Pd-Cu2O has good hydrogen evolution activity, which promotes the generation of oxygen by improving the redox efficiency of the overall reaction, thus increasing the yield of active intermediates (OH•) to promote the reduction of S2O82-. Specially, this is an effective attempt to use the hydrogen evolution reaction (HER) to accelerate the ECL emission of the S2O82- system. In addition, a short peptide ligand (NARKFYKGC, NFC) was developed to implement the targeted immobilization of antibodies, which can specifically bind to the Fc fragment of antibodies, thereby avoiding the occupation of the antigen binding site (Fab fragment). The introduction of NFC not only improves the binding efficiency of antibodies but also protects its bioactivity, thus significantly improving the sensitivity of the biosensor. Based on these strategies, the proposed biosensor provides a new perspective for the applications of metal NCs in ECL systems.

Peptide-Based Biosensor with a Luminescent Copper-Based Metal-Organic Framework as an Electrochemiluminescence Emitter for Trypsin Assay.

A copper-based metal-organic framework (JUC-1000) has emerged as a promising electrochemiluminescence (ECL) emitter in the domains of bioanalysis and immunoassay. Herein, a highly efficient signal "on-off" peptide-based biosensor was constructed for trypsin (TPN) assay. JUC-1000 synthesized using an organic ligand of H4BDPO was functionalized as the ECL emitter, whose cathodic ECL behavior in aqueous media was first investigated using potassium persulfate (K2S2O8) as the coreactant. To further amplify the ECL signal, highly catalytic Ag@CeO2 nanoparticles were fabricated as both a substrate and an coreaction accelerator, which can efficiently catalyze the reduction of S2O82- to generate more sulfate anion radicals (SO4•-) for ECL enhancement, thereby generating strong and stable ECL signals in a "signal on" state. The functionalized JUC-1000 emitter was connected to the Ag@CeO2 sensing layer though a heptapeptide (HWRGWVC, HGC), and TPN as the target can specifically cleave the carboxyl side of arginine residues in HGC, leading to the release of emitters in a "signal off" state. Based on the efficient signal-switching, the biosensor exhibited linear ECL responses to the added TPN concentration, realizing sensitive detection of TPN in 10 fg/mL to 100 ng/mL with a limit of detection of 3.46 fg/mL. This work proposed an attractive orientation for the fundamental research of applying transition metal-organic frameworks as ECL emitters in bioanalysis and immunoassay.

Rational design of the Z-scheme hollow-structure Co9S8/g-C3N4 as an efficient visible-light photocatalyst for tetracycline degradation.

The development of photocatalysts with high catalytic activity that are capable of full utilization of solar energy is a challenge in the field of photocatalysis. Accordingly, in the present study, an efficient Z-scheme cage-structured Co9S8/g-C3N4 (c-CSCN) photocatalyst was constructed for the degradation of tetracycline antibiotics under visible-light irradiation. The Z-scheme charge-transfer mechanism accelerates the separation of photogenerated charge carriers and effectively improves photocatalytic activity. Moreover, c-CSCN has a hollow structure, allowing light to be reflected multiple times inside the cavity, thereby effectively improving the utilisation efficiency of solar energy. As a result, the photocatalytic activity of c-CSCN is 1.5-, 2.5-, and 5.8-times higher than those of sheet-type Co9S8/g-C3N4 (s-CSCN), c-Co9S8, and g-C3N4, respectively, for the degradation of tetracycline. c-CSCN maintains favourable photocatalytic activity over five consecutive degradation cycles, demonstrating its excellent stability. In addition, c-CSCN performs efficient tetracycline removal in different water substrates. Moreover, c-CSCN exhibits excellent ability to remove tetracycline under direct natural sunlight. This work fully demonstrates that c-CSCN has high catalytic activity and the potential for practical application as a wastewater treatment material.

Electrochemical assays for determination of H2O2 and prostate-specific antigen based on a nanocomposite consisting of CeO2 nanoparticle-decorated MnO2 nanospheres.

A nanocomposite consisting of CeO2 nanoparticle-decorated MnO2 nanospheres (CeO2@MnO2) was synthesized for the first time via a hydrothermal method. CeO2@MnO2 was exploited to construct an electrochemical assays for detecting H2O2 and prostate-specific antigen (PSA) with square wave voltammetry (SWV). The electrochemical results proved that CeO2@MnO2 owned a better electrocatalytic effect towards H2O2 reduction than pure MnO2 NS and CeO2 NP due to the synergistic effect between MnO2 NS and CeO2 NP. Under optimized conditions, CeO2@MnO2-based assay can be applied to detect H2O2 in the range 1 to 3.0 × 103 µmol L-1. The label-free electrochemical immunoassay based on CeO2@MnO2 displayed linearly with concentrations of PSA from 0.005 to 50.0 ng mL-1. The electrochemical assays also possessed acceptable sensitivity, selectivity, and stability. The study showed that CeO2@MnO2 hold great potential as a biosensing platform and the clinical determination of tumor markers in human serum. Graphical abstract A nanocomposite consisting of CeO2 nanoparticles decorated MnO2 nanospheres (CeO2 @MnO2) was firstly synthesized via a hydrothermal method. CeO2@MnO2 was firstly exploited to construct electrochemical assays for detecting H2O2 and prostate-specific antigen (PSA) with square wave voltammetry (SWV), respectively. The electrochemical results proved that CeO2@MnO2 owned better electrocatalysis towards H2O2 reduction than pure MnO2 NS and CeO2 NP due to the synergistic effect between MnO2 NS and CeO2 NP. Under optimized conditions, CeO2@MnO2 based assay relative to the H2O2 system can be applied to detect H2O2 with range from 1 to 3.0 × 103 µmol L-1. The label-free electrochemical immunoassay based on CeO2@MnO2 relative to the H2O2 system displayed linearly with concentrations of PSA from 0.005 to 50.0 ng mL-1. The electrochemical assays also possessed acceptable sensitivity, selectivity and stability. The study showed that CeO2@MnO2 hold great potential for biosensing platform and the clinic determination of tumor markers in human serum.

A chemiluminescence assay for determination of lysozyme based on the use of magnetic alginate-aptamer composition and hemin@HKUST-1.

Lysozyme aptamer-functionalized magnetic alginate hydrogel was prepared for separation and enrichment of lysozyme. Luminol-labeled aptamer was used as a signal tag, and the signal tag was adsorbed on magnetic carboxylated carbon nanotubes based on the π-interaction. When lysozyme was added, the aptamer specifically binds to the lysozyme, causing the signal tag to detach from the magnetic carboxylated carbon nanotubes. When the aptamer/lysozyme complex bound to the complementary single strand of aptamer on the hemin@HKUST-1, lysozyme was released. The released lysozyme can be recombined with the signal tag adsorbed on the magnetic carboxylated carbon nanotube, allowing more signal tag to be dispersed into the solution. Determination of lysozyme was achieved by releasing the luminol-labeled aptamer to generate a chemiluminescence signal at a wavelength of 425 nm. It was proved by experiments that the synthesized hemin@HKUST-1 had a strong catalytic effect on the luminol-NaOH-H2O2 system. The chemiluminescence signal was increased nearly 100 times. The complementary pairing allowed the luminol to be immobilized on the surface of hemin@HKUST-1. The generation and consumption of short-lived reactive oxygen species were concentrated on the surface of the MOFs, which improves the chemiluminescence efficiency. The introduction of hemin@HKUST-1 and DNA solved the defects of chemiluminescence analysis. The chemiluminescence assay was able to detect lysozyme with linear range of 1.05 × 10-6 U∙mg-1 (6.00 × 10-13 mol∙L-1)-1.25 × 10-2 U∙mg-1 (7.14 × 10-9 mol∙L-1); the detection limit was 3.50 × 10-7 U∙mg-1 (2.00 × 10-13 mol∙L-1) (R2 = 0.99). The recovery of lysozyme in spiked saliva samples was 97.4-102.8%. Graphical abstract Schematic presentation of chemiluminescence assay. Lysozyme (Lys) was captured by aptamer-modified magnetic sodium alginate (M-Alg-Apt); Glycine (pH = 2) as eluent for Lys. Luminol-modified Apt (Apt-luminol) as signal tag; magnetic carbon nanotubes (MCNTs) as adsorption matrix; cDNA was complementary to Apt; hemin@HKUST-1 as catalyst.

Triple Amplification of 3,4,9,10-Perylenetetracarboxylic Acid by Co2+-Based Metal-Organic Frameworks and Silver-Cysteine and Its Potential Application for Ultrasensitive Assay of Procalcitonin.

In this work, a triple-amplified biosensor with a bioactivity-maintained peculiarity was constructed for quantitative procalcitonin (PCT) detection. As everyone knows, a strong electrochemiluminescence (ECL) signal is the premise to ensure high sensitivity for trace target detection. Hence, a valid tactic was developed to achieve signal amplification of luminophor by using Co2+-based metal-organic frameworks (ZIF-67) and silver-cysteine (AgCys). The ZIF-67 particles, which have more atomically dispersed Co2+, could play the role of a co-reaction accelerator to catalyze S2O82- to generate abundant Co3+ and sulfate radical anions (SO4•-). Afterward, a mass of Co3+ was reduced to more hydroxyl radicals (OH•) by H2O, thus ulteriorly reducing S2O82- to generate more SO4•-. Remarkably, S2O82- was reduced to SO4•- continuously with the recycling of Co2+ and Co3+, which realized an effective signal amplification. Meanwhile, the AgCys complex with superior catalysis and biocompatibility was prepared to further improve the ECL signal and maintain the bioactivity of the biomolecule. Furthermore, HWRGWVC, a heptapeptide that was used for combining the Fc fragments of an antibody by Au-S bonding to achieve the fixed point fixation, could not only maintain bioactivity of an antibody but also improved its incubation efficiency, thus further enhancing biosensor sensitivity. Under optimum conditions, the proposed biosensor realized highly sensitive assay for PCT with a wide dynamic range from 10 fg/mL to 100 ng/mL and a detection limit as low as 3.67 fg/mL. With superior stability, selectivity, and repeatability, the prepared biosensor revealed immense potential application of ultrasensitive assay for PCT in human serum.

Novel Chemiluminescence Sensor for Thrombin Detection Based on Dual-Aptamer Biorecognition and Mesoporous Silica Encapsulated with Iron Porphyrin.

Thrombin is a marker of blood-related diseases, and its detection is of great significance in the fields of medical and biological research. Herein, a novel chemiluminescence (CL) sensor for thrombin detection was prepared based on dual-aptamer biorecognition and mesoporous silica encapsulated with iron porphyrin. Mesoporous silica encapsulated with hematin by aptamer1 (Apt1/hematin/M-SiO2) and magnetic microspheres modified with aptamer2 (Apt2/NH2-MS) were successfully prepared, and the two materials were used to construct a CL sensor to detect thrombin. Primarily, Apt2/NH2-MS is used for pretreatment separation of thrombin samples by the biorecognition effect between the aptamer (Apt2) and target (thrombin). Then, thrombin/Apt2/NH2-MS is again recognized with Apt1 on the surface of Apt1/hematin/M-SiO2 and Apt1/thrombin/Apt2/NH2-MS is formed, so dual-aptamer biorecognition is realized. Meanwhile, the generated Apt1/thrombin/Apt2/NH2-MS makes Apt1 shed off the surface of M-SiO2 and release hematin. The released hematin can catalyze the luminol-H2O2 CL reaction. Therefore, a sandwich-type CL sensor was constructed based on dual-aptamer biorecognition and hematin catalysis for the detection of thrombin. The sensor has a linear range of 7.5 × 10-15 to 2.5 × 10-10 mol·L-1 and a detection limit of 2.2 × 10-15 mol·L-1 and also exhibits excellent selectivity, reproducibility, and stability. The sensor was successfully used for the detection of thrombin in serum samples, which makes it possible to apply the sensor in the detection of thrombin in actual samples.

A "signal-on" chemiluminescence biosensor for thrombin detection based on DNA functionalized magnetic sodium alginate hydrogel and metalloporphyrinic metal-organic framework nanosheets.

A "signal-on" chemiluminescence biosensor was established for detecting thrombin. The thrombin aptamer1-functionalized magnetic sodium alginate (Malg-Apt1) hydrogel was synthesized by physical interaction between sodium alginate and Ca2+, and it was used in the biosensor for separating and enriching thrombin. Ethylenediamine tetraacetic acid (EDTA) was used to chelate with Ca2+ to dissolve the hydrogel and release thrombin. A metalloporphyrinic metal-organic framework nanosheet, named as Cu-TCPP(Co) MOFs, was prepared as signal amplification strategy. Cu-TCPP(Co) MOFs/Au-ssDNA (ssDNA: single-strand DNA) was synthesized for controllable further amplification of chemiluminescent signal. The thrombin aptamer2-functionalized magnetic carbon nanotubes (MCNTs-Apt2) were used as a matrix, and Cu-TCPP(Co) MOFs/Au-ssDNA was adsorbed on the MCNTs by the complementary pairing of the partial bases between ssDNA and Apt2. Compared with ssDNA, Apt2 has a stronger interaction with thrombin. Therefore, thrombin can trigger the release of Cu-TCPP(Co) MOFs/Au-ssDNA to achieve signal amplification. Under the optimal conditions, the biosensor could detect thrombin as low as 2.178 × 10-13 mol/L with the range from 8.934 × 10-13 to 5.956 × 10-10 mol/L and exhibited excellent selectively. Moreover, the "signal-on" chemiluminescence biosensor showed potential application for the detection of thrombin in body fluids.

Enhanced electrochemiluminescence of luminol based on Cu2O-Au heterostructure enabled multiple-amplification strategy.

Herein, a credible construction strategy to improve electrochemiluminescence (ECL) of luminol was developed based on Cu2O-Au heterostructures. Summarily, gold nanoparticles (AuNPs) were anchored on surface of Cu2O nanocube (Cu2O@AuNPs) by spontaneous reduction reaction. Then, luminol molecules were concentrated on Cu2O@AuNPs using L-Cysteine (Cys) as covalent linkage to build the composite emitter (Cu2O@AuNPs-Cys-luminol). The enhancement mechanism was realized by following aspects: (I) Cu2O@AuNPs worked as electrocatalyst for glucose to generate coreactant of H2O2 in situ, avoiding the instability of direct addition of H2O2. (II) luminol molecules were firmly attached on Cu2O@AuNPs to achieve centralized and strong luminescence at low consumption. (III) Cys acted as an intramolecular coreactant and directly linked to luminol to increase luminous efficiency. To validate the effectiveness, a sandwiched immunoassay was built using concanavalinA (ConA) as analyte. Electroreduced graphene film as substrate provided phenoxy-derivatized dextran (DexP) with abundant binding sites and improved conductivity. To improve the specificity, DexP was used to identify ConA via the specific carbohydrate-ConA interaction. Then, Cu2O@AuNPs-Cys-luminol was modified on electrode as ECL signal indicator. The ECL immunosensor achieved determination of ConA with low detection limit of 2.9 × 10-5 ng/mL and excellent stability of continuous potential scan for 8 cycles. Experimental results demonstrated that the proposed construction strategy made considerable progress in ECL efficiency and stability of luminol. The creational pattern of construction strategy achieves high detection capabilities to ConA and expands the applicability of luminol in ECL system. It is expected to have more potential application value in immunoassay with universality.

Rapid fabrication of electrode for the detection of alpha fetoprotein based on MnO2 functionalized mesoporous carbon hollow sphere.

The concentration of alpha-fetoprotein (AFP) rises greatly in patients with liver cancer and it is a challenge to construct a sensitive AFP detection method with wide range. Therefore, an easy and label-free sensing electrochemical platform for AFP detection with wide concentration range had been designed in this work. Firstly, MnO2 functionalized mesoporous carbon hollow sphere (MCHS@MnO2) with optimal performance was synthesized by regulating experimental conditions and characterized by scanning electron microscope (SEM), high resolution transmission electron microscopy (HRTEM), x-ray diffraction (XRD), electrochemical impedance spectroscopy (EIS), and cyclic voltammetry (CV), etc. Then, it was immobilized on glassy carbon electrode (MCHS@MnO2/GCE) to build an immunosensor for the detection of AFP. The MCHS@MnO2/GCE can catalyze decomposition of H2O2 to generate electrochemical signal, and the signal will decrease after capturing AFP. Due to good electrocatalytic activity of MCHS@MnO2 to H2O2, the immunosensor achieved indirect detection of AFP with wide sensing range from 0.10 ng mL-1 to 420 ng mL-1 and a detection limit of 0.03 ng mL-1. Furthermore, the method had been proven to be satisfactory selectivity and reproducibility, and it was successfully applied to determine the content of AFP in human serum samples with satisfactory results. This method is expected to be used for early diagnosis and prognosis examination of liver cancer patients.

A nanocomposite prepared from bifunctionalized ionic liquid, chitosan, graphene oxide and magnetic nanoparticles for aptamer-based assay of tetracycline by chemiluminescence.

A nanocomposite was prepared from a bifunctionalized ionic liquid, chitosan on magnetic nanoparticle-modified graphene oxide (IL/Chit@MGO). It was used in a chemiluminescencc (CL) assay for tetracycline. The materials were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray powder diffraction, nitrogen adsorption-desorption isotherm, vibrating sample magnetometry and zeta potentials. Subsequently, a tetracycline-binding aptamer (TC-Apt) acting as a recognition element, and G-quadruplex DNAzyme (G-DNAzyme) acting as a signal amplification component were modified on IL/Chit@MGO. So, the bifunctional G-DNAzyme/TC-Apt/IL/Chit@MGO was prepared. The IL/Chit@MGO is found to possess excellent loading capability for TC-Apt. This is attributed to the large specific surface and abundant charge on the surface of IL/Chit@MGO. The composite was used to construct a CL assay for tetracycline. Tetracycline binds to TC-Apt and causes the release of the G-DNAzyme. The latter catalyzes the CL of luminol-H2O2 CL system at pH 7.4. Under optimized conditions, the blue CL at the emission wavelength of 425 nm increases linearly in the 0.16 pM to 2.0 nM concentration range, and the detection limit is 21 fM (at 3σ). The assay is selective, reproducible and stable. The assay was applied to tetracycline detection in practical samples. The apparent recoveries are 98.0% to 101.3% for the milk sample and 97.0% to 102.2% for the water sample. Graphical abstractG-quadruplex DNAzyme (G-DNAzyme) and tetracycline aptamer (TC-Apt) bifunctionalized ionic liquid/chitosan@magnetic graphene oxide (IL/Chit@MGO) was prepared. The nanocomposite was used to construct a chemiluminescence (CL) assay for tetracycline.