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BIOSENSORS & BIOELECTRONICS · 2017 · IF 7.78

Multiple amplified enzyme-free electrochemical immunosensor based on G-quadruplex/hemin functionalized mesoporous silica with redox-active intercalators for microcystin-LR detection

Cuifen Gan,Bingfeng Wang,Junying Huang,Aori Qileng,Zuyu He,Hongtao Lei,Weipeng Liu,Yingju Liu

A novel multiple amplified enzyme-free immunosensor was developed for competitive immunoassay of microcystin-LR (MC-LR). Classical electrochemical immunosensors usually employ enzymes as biocatalysts to afford amplified signals, but the proteolytic degradation and poor stability are still crucial problem. Herein, monodisperse core-shell mesoporous silica (SiO 2 @MSN)-functionalized DNAzyme concatamers were synthesized to load hemin and methylene blue (MB) as the mimic enzyme . Firstly, the surface of SiO 2 @MSN was conjugated with secondary antibody as the recognition of MC-LR antibody and with a DNA strand as the initiator. Two auxiliary DNA strands were then selected for the in-situ propagation to form a double-helix DNA through hybridization chain reaction (HCR), forming numerous DNAzymes (G-quadruplex/hemin) after the addition of hemin. Secondly, MB was inserted into the formed double-helix DNA, and also loaded in the brush-like structure of mesoporous SiO 2 @MSN. The molecular docking study showed that electrons can transfer more effectively with π-π stack of hemin/G-quadruplex and intercalation of MB/DNA, thus the catalytic ability of DNAzymes can be greatly improved. With the aid of MB, DNAzymes can catalyze the reduction of H 2 O 2 to produce the electrochemical signal. This enzyme-free electrochemical immunosensor can successfully detect MC-LR in a range of 0.5   ng/L and 25   μg/L with a detection limit of 0.3   ng/L. This stable, sensitive and selective nonenzymatic electrochemical immunoassay shows promise for applications in food and environmental monitoring.

研究领域: 分析化学 · 环境科学 · 生物技术 · 纳米技术
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MICROCHIMICA ACTA · 2016 ·Vol. 183 ·期 11 ·2965-2971 · IF 4.831

Toehold-aided DNA recycling amplification using hemin and G-quadruplex reporter DNA on magnetic beads as tags for chemiluminescent determination of riboflavin

Meng Yan,Hun Xu,Zhang Yue,Luo Xiliang

A toehold-aided DNA recycling amplification technology was developed based on the combination of toehold-aided DNA recycling and the hemin/G-quadruplex label. The dsDNA formed between aptamer and DNA1 was first immobilized on magnetic beads. On addition of target analyte (exemplified here for riboflavin), the aptamer-riboflavin complex is formed and DNA1 is released by the beads. After magnetic separation, the supernatant containing the released DNA1 is added to a solution containing the hairpin capture DNA on magnetic beads. DNA1 will hybridize with the hairpin capture DNA via toehold binding and branch migration. This process will open the hairpin structure, and an external toehold is formed in the newly formed dsDNA. On addition of reporter DNA containing the G-quadruplex, it will interact with the formed dsDNA via toehold binding and branch migration, resulting in the releasing of DNA1 and capturing of reporter DNA on the magnetic beads. The released DNA1 will bind to another hairpin capture DNA which can start another round of DNA1 recycling. Chemiluminescence (CL) is generated by the G-quadruplex-hemin-luminol CL reaction system. Under optimal conditions, the calibration plot is linear in the 0.1 to 700 nM riboflavin concentration range, with a 30 pM detection limit (at a signal-to-noise ratio of 3). The method was successfully applied to the quantitation of riboflavin in spiked urine samples. Toehold-aided DNA recycling coupled with hemin G-quadruplex for target detection.

研究领域: 生物化学 · 分析化学 · 分子生物学
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TALANTA · 2021 · IF 6.057

Peptide nucleic acid-assisted colorimetric detection of single-nucleotide polymorphisms based on the intrinsic peroxidase-like activity of hemin-carbon nanotube nanocomposites

Mengjia Xu,Shu Xing,Yang Zhao,Chao Zhao

Here, taking the advantage of single-stranded (ss) DNA specific nuclease (S1) and peptide nucleic acid (PNA), we demonstrated a novel, rapid, and label-free colorimetric nanosensor for the sensitive and accurate detection of SNPs based on the intrinsic peroxidase-like activity of hemin-functionalized single-walled carbon nanotubes (hemin-SWCNTs). PNA, a man-made mimic of DNA with extraordinary stability toward enzymatic degradation, can effectively protect DNA in the fully matched DNA/PNA duplexes from nuclease digestion. While the DNA in DNA/PNA duplexes containing a mismatch can be cleaved into small fragments. This difference can be visually monitored from the specific color change of TMB/H 2 O 2 system by employing the peroxidase activity of hemin-SWCNTs because of its different aggregation states responding to ssPNA or DNA/PNA duplex. Under optimized conditions, the SNPs in the human tumor suppressor gene TP53 have been successfully genotyped in a linear range of 50–1000 nM with a detection limit of 0.11 nM. Moreover, this platform can effectively discriminate a series of single-base mismatches. This assay avoids the assistance of sophisticated instruments and complicated modifications of probes or nanomaterials , and function well for both cell lysate samples and PCR amplicons from standard cell lines, implying its potential practical applications for bioanalysis and biosensors.

✓ 已验证 产品关联 DOI PubMed 高影响因子
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FOOD CHEMISTRY · 2025 · IF 9.8

Fabrication of dBSA@Hemin@Ag nanozyme based biosensor and application for the assessment of longitudinal concentration changes of choline in human breast milk

Peng Liu,Huimin Deng,Huijun Chen,Jiaoyang Li,Liangyi Hong,Huimin Zou,Xingyan Zhou,Hian Kee Lee,Zhenzhen Huang

Choline in human breast milk is crucial for infant neurodevelopment. However, choline concentration variation in human breast milk remains understudied. In this work, a colorimetric biosensor of bovine serum albumin@Hemin@Ag nanozyme was developed, leveraging its peroxidase-like activity to oxidize 2,2′-azino-bis(3ethylbenzo-thiazoline-6-sulfonic acid) diammonium salt in the presence of hydrogen peroxide generated from choline oxidation by choline oxidase. Under the optimal conditions, a wide linear range (1–500 μM) and a low limit of detection (0.55 μM) were obtained for choline quantification. Longitudinal analysis of paired human breast milk samples revealed a significant decline in choline level from 15- to 90-d postpartum, influenced by maternal diet during pregnancy and pre-pregnancy body mass index. This research not only provides a robust tool for evaluating breast milk's quality, but also underscores the importance of diet during pregnancy and body mass index management for lactating mothers.

研究领域: 生物传感器 · 纳米技术 · 分析化学 · 母婴健康 · 营养生物化学
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MICROCHIMICA ACTA · 2020 ·Vol. 187 ·期 11 ·1-10 · IF 6.232

Colorimetric and fluorescent dual-identification of glutathione based on its inhibition on the 3D ball-flower shaped Cu-hemin-MOF’s peroxidase-like activity

Chen Xiaolong,Wang Xianfeng,Cao Gaihua,Wu Yawen,Luo Huibo,Ji Zhong,Shen Caihong,Huo Danqun,Hou Changjun

Hemin as the organic linker ligand and Cu (II) as the metal center were applied to prepare a copper-metal-organic framework (Cu-hemin-MOF) via one-step hydrothermal method. Characterization using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), and X-ray powder diffraction (XRD) demonstrate that the acquired Cu-hemin-MOF possesses the appearance of 3D ball-flower shape with the existence of C, N, O, Fe, and Cu on the surface. Further study found that this 3D ball-flower shaped Cu-hemin-MOF exhibits peroxidase-like activity, which can catalyze the peroxidase substrate of o-phenylenediamine (OPD) to generate 2,3-diaminophenazine (DAP) in the presence of H 2 O 2 . DAP has a yellow color and also emits a strong fluorescence when excited by ultraviolet light. Interestingly, Cu-hemin-MOF’s peroxidase-like activity can be strongly inhibited by glutathione (GSH). In view of this, a dual readout (fluorescence and colorimetry) was proposed to detect GSH for the first time. Under optimal conditions, the proposed method exhibits good linear relationship between the signal response (fluorescence and colorimetry) and the concentration of GSH, and low limits of detection (LOD) of 2.3 and 26.6 nM, respectively. Graphical abstract

研究领域: 材料科学 · 分析化学 · 生物化学
✓ 已验证 DOI PubMed 高影响因子
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Analytical Methods · 2024 · IF 2.7

A colorimetric strategy and smartphone-based test strip for the detection of glucose based on the peroxidase activity of a hemin-derived nanozyme

Ziqiang Yuan,Meizhen Fu,Xin Wang,Meng Wang,Yue Wei,Yanna Sun,Qingfeng Zhang,Yunyi Zhang,Bo Zhang

Nanozymes are nanomaterials with catalytic properties similar to enzymes and offer advantages over natural enzymes such as lower cost, ease of storage, and convenience for large-scale preparation. Thus, they have received increasing attention and are extensively applied in fields such as chemistry, sensing, food, environment, and medicine. Herein, a hemin-derived nanozyme (Hemin-CDs) was prepared using hemin as the precursor and used to substitute the natural horseradish peroxidase (HRP) for colorimetric detection. The prepared Hemin-CDs exhibit excellent water dispersibility, stability and superior peroxidase-like activity. They catalyze the oxidative coupling of colorless 4-aminoantipyrine (4-AAP) with N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methylaniline sodium salt (TOOS) in the presence of H2O2, forming a purple quinone diimine dye with an absorption peak at 554 nm, which enables the quantification of H2O2 by measuring the absorbance. Quantitative detection of glucose was further demonstrated under physiological conditions. The detection of H2O2 and glucose takes only 10 minutes, with linear ranges of 1–150 μM and 2.5–300 μM, and LODs of 0.867 μM and 1.026 μM, respectively. Glucose in human serum was successfully detected with satisfactory recoveries (87.5–104.0%). Furthermore, a test strip was developed, and a smartphone was utilized to recognize the color of the test position, enabling rapid and accurate quantification of glucose concentrations within 5 minutes, which promises to enhance the accessibility and convenience of glucose testing.

研究领域: 生物化学 · 医学诊断 · 纳米技术
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Analytical Methods · 2017 ·Vol. 10 ·期 1 ·51-58 · IF 1.9

An electrochemiluminescence sensor based on CdSe@CdS functionalized MoS2 and hemin/G-quadruplex-based DNAzyme biocatalytic precipitation for sensitive detection of Pb(II)

Xiao-Li Du,Tian-Fang Kang,Li-Ping Lu,Shui-Yuan Cheng

A novel electrochemiluminescence (ECL) biosensor based on CdSe@CdS quantum dots (QDs) functionalized MoS2 modified electrode was developed for sensitive detection of Pb(II) using HRP-mimicking DNAzyme biocatalytic precipitation (BCP) for ECL signal quenching. Polycationic poly(diallyldimethylammonium chloride) (PDDA) and negatively charged QDs were adsorbed on the surface of MoS2, in sequence, to form the MoS2–QDs composites, which were employed as a matrix for the immobilization of the G-rich DNA strand, i.e., T30695. The MoS2–QDs were characterized by UV-vis absorption spectroscopy, fluorescence spectroscopy, energy dispersive spectrometry, transmission electron microscopy, and field emission scanning electron microscopy. CdSe@CdS QDs can produce an ECL signal using 0.1 M K2S2O8 as a co-reactant. T30695 can combine with Pb(II) ions to form a stable parallel G-quadruplex. The G-quadruplex can further combine with hemin to form DNAzyme, which can catalyze H2O2 and oxidize 4-chloro-1-naphthol (4-CN) to produce an insoluble precipitation of benzo-4-chlorohexadienone on the surface of the sensor. When the concentration of Pb(II) in the solution was increased, more of DNAzyme was generated and subsequently more 4-CN molecules were oxidized catalytically, leading to a decrease of the ECL signal. Using this quenching strategy, the ECL signal of the biosensor was linearly dependent on the logarithm of the Pb(II) concentration ranging from 1.0 × 10−15 to 1.0 × 10−11 M with a detection limit of 0.98 f M. The stability and selectivity of the sensor were also investigated. The sensor was applied in the determination of Pb(II) in the real water samples and satisfactory results were obtained.

CAS / MDL: 16009-13-5
研究领域: 分析化学 · 环境科学 · 纳米技术 · 生物传感器
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY · 2018 ·Vol. 410 ·期 8 ·2211-2219 · IF 3.307

Photoinduced electron transfer from polymer-templated Ag nanoclusters to G-quadruplex-hemin complexes for the construction of versatile biosensors and logic gate applications

Qu Fei,Mao Beibei,Xue Fangfang,Xia Lian,You Jinmao,Song Cuihua

In this paper, fluorescent Ag nanoclusters (Ag NCs) templated by hyperbranched polyethyleneimine (PEI) are utilized as a versatile probe through the photoinduced electron transfer (PET) between PEI-Ag NCs and G-quadruplex-hemin complexes. In the presence of hemin and target molecule, the specific conjugation with its aptamer induces the conformational change of the DNA sequence, releasing the G-quadruplex sequence part. Once the G-quadruplex-hemin complexes are introduced, electron transfer from the PEI-Ag NCs to G-quadruplex-hemin complexes occurs, resulting in fluorescence quenching. Through changing the sensing DNA sequence, this novel PET system enables the specific detection of target DNA and adenosine triphosphate (ATP) with the wide linear range of 1–200 nM and 5–500 nM, respectively, and the corresponding limit of detection as low as 0.3 nM for target DNA and 1.5 nM for ATP. In addition, the proposed method is successfully applied to the determination of ATP in human serum samples with satisfactory recoveries, and a logic gate is fabricated using target molecules and hemin as inputs and the fluorescence signal of PEI-Ag NCs as an output.

研究领域: 生物化学 · 纳米生物学 · 分析化学
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JOURNAL OF ELECTROANALYTICAL CHEMISTRY · 2017 · IF 3.012

Sandwich-like electrochemiluminescence aptasensor based on dual quenching effect from hemin-graphene nanosheet and enzymatic biocatalytic precipitation for sensitive detection of carcinoembryonic antigen

Jiu-Jun Yang,Jun-Tao Cao,Yu-Ling Wang,Hui Wang,Yan-Ming Liu,Shu-Hui Ma

A new and simple sandwich-like electrochemiluminescence (ECL) aptasensor for carcinoembryonic antigen (CEA) assay was fabricated based on the dual quenching effect from hemin-graphene (H-rGO) nanosheet and enzymatic biocatalytic precipitation (BCP) on the Au-CdS nanocomposites-based ECL system. In this aptasensor platform, flower-like spherical Au-CdS nanocomposites were used as ECL luminophores and exhibit a strong ECL signal. The rGO nanosheet was used as a supporter to immobilize hemin molecules via π-π stacking interactions. Due to the steric hindrance and quenching effect of rGO, the ECL intensity decreased by the construction of the sandwich “CEA aptamer I (NH 2 -DNA)-CEA-aptamer II” (H-rGO-aptamer II) mode. In the process of BCP, the ECL intensity further decreased because the hemin with intrinsic peroxidase-like catalytic activity could oxidize the 4-chloro-1-naphthol (4-CN) to produce an insoluble precipitation on the sensor. Using this dual quenching strategy, the prepared aptasensor exhibits a linear range from 0.8   pg/mL to 4   ng/mL and a detection limit of 0.28   pg/mL. This ECL aptasensor has simple design and undemanding in operation and was utilized to determine the content of CEA in complex samples with recoveries of 95.0% to 115.8%. Moreover, no any chemical modification of aptamer was required, suggesting that the proposed ECL aptasensor could be applied for the detection of diverse proteins just by altering the aptamer sequence.

CAS / MDL: 107-21-1 34819-78-8
研究领域: 分析化学 · 生物传感器 · 纳米技术 · 癌症诊断
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BIOSENSORS & BIOELECTRONICS · 2019 · IF 9.518

Highly electrocatalytic biosensor based on Hemin@AuNPs/reduced graphene oxide/chitosan nanohybrids for non-enzymatic ultrasensitive detection of hydrogen peroxide in living cells

Wenjing Wang,Huabiao Tang,Yuan Wu,Yinli Zhang,Zhaohui Li

A non-enzymatic and highly electrocatalytic H 2 O 2 biosensor was proposed by using a novel electrode composed of hemin-capped biomineralized gold nanoparticles ( [email protected] ), reduced graphene oxide (rGO) and chitosan (CS). Owing to the excellent electrocatalytic activity of [email protected] and great conductivity of rGO, this developed biosensor is capable of ultrasensitive measuring H 2 O 2 in real-time. In addition, the introduction of CS resulted in a film on the electrode, which can enhance the stability by protecting the biosensor from external disturbances . Taking advantages of the peroxidase-like activities of the nanohybrids , the developed electrode exhibited outstanding electrochemical performance toward H 2 O 2 with high selectivity , fast response, superior sensitivity and good stability. More importantly, the lower determination limit of 9.3 nM and wider linear ranges of 5 orders of magnitude enable this biosensor to detect H 2 O 2 releasing from living Hela cells accurately. This work provides tremendous potential for real-time monitoring the secretion of H 2 O 2 in living cells.

CAS / MDL: 50-81-7
研究领域: 生物传感器 · 电化学 · 纳米技术 · 分析化学 · 生物医学工程
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JOURNAL OF THE ELECTROCHEMICAL SOCIETY · 2021 ·Vol. 168 ·期 12 ·127501 · IF 4.316

Fe-Hemin-Metal Organic Frameworks/Three-Dimensional Graphene Composites with Efficient Peroxidase-Like Bioactivity for Real-Time Electrochemical Detection of Extracellular Hydrogen Peroxide

Shiying Zhou,Human Sun,Xianfeng Wang,Peng Lu,Danqun Huo,Jiawei Li,Xiaogang Luo,Changjun Hou

Real-time detection of extracellular hydrogen peroxide is important for dynamic monitoring of diseases and cytopathological research. Here, a novel composite of Fe-hemin-metal organic frameworks/three-dimensional graphene (Fe-hemin MOFs/3DG) was prepared by embedding hemin into amino-contained Fe-MOFs, then decorated with 3DG. The obtained Fe-hemin MOFs/3DG possessed efficient peroxidase-like bioactivity and could serve as an effective catalyst for construction of H2O2 electrochemical sensor. The electrochemical results show that the proposed sensor based on Fe-hemin MOFs/3DG has excellent catalytic activity for H2O2. With a linear range was 10−12080 μM and LOD was 0.34 μM, this sensor showed favorable selectivity, repeatability and stability, which could be used to detect H2O2 in real serum samples. Most importantly, this sensor realized the real-time determination of H2O2 released by A549 cells and possessed good biocompatibility. The outstanding electrochemical performance mainly benefited from the unique porous structure of MOFs, which could effectively protect the active center of hemin, and the introduction of 3DG greatly improved the conductivity of Fe-hemin MOFs. Therefore, the Fe-hemin MOFs/3DG could be a promising probe for real-time dynamic monitoring of H2O2.

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Biomaterials Science · 2022 ·Vol. 10 ·期 23 ·6804-6817 · IF 7.59

A label-free electrochemical aptasensor based on platinum@palladium nanoparticles decorated with hemin-reduced graphene oxide as a signal amplifier for glypican-3 determination

Guiyin Li,Wenzhan Li,Shengnan Li,Xinhao Li,Xiaoqing Yao,Wen Xue,Jintao Liang,Jiejing Chen,Zhide Zhou

Glypican-3 (GPC3) is a membrane-associated proteoglycan that is specifically upregulated in hepatocellular carcinoma (HCC) and has become one of the most promising biomarkers closely related to the occurrence and development of HCC. In this work, platinum@palladium nanoparticles decorated with hemin-reduced graphene oxide (H-rGO-Pt@Pd NPs) were used not only as a support for GPC3 aptamer (GPC3Apt) immobilization, but also as a new redox nanoprobe in electrochemical analysis for the determination of GPC3. The electrochemical aptasensor involved a reaction cell with a three-electrode system, and the differential pulse voltammetry (DPV) technique was adopted. In the presence of GPC3, the formed GPC3Apt–GPC3 complexes had stable structures and were cleaved from the electrode surface, leading to more electroactive H-rGO-Pt@Pd NPs repelling freely from the GPC3Apt/H-rGO-Pt@Pd NPs and thus to the increase of the oxidation peak current of hemin in H-rGO-Pt@Pd NPs. Under optimal conditions and a working voltage of +700 mV (vs. Ag/AgCl), the label-free electrochemical GPC3 aptasensor showed superior performance with a wider concentration linear range (0.001–10.0 μg mL−1), a lower limit of detection (LOD) (0.181 ng mL−1, S/N = 3), a higher sensitivity (0.0446 μA μM−1 cm−2) and good selectivity. Furthermore, the fabricated aptasensor was applied to GPC3 determination in human serum samples with satisfactory recoveries of 94.3%–119% and RSDs of 0.15%–5.78%. The current work provides a flexible approach for the rapid and sensitive analysis of GPC3 and has a broad application prospect in the diagnosis of HCC.

研究领域: 生物医学工程 · 分析化学 · 肿瘤学
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