Journal of Biomedical Nanotechnology
·
2021
·Vol. 17
·期 12
·2444-2454
·
IF 4.099
Li Guiyin,Li HaiMei,Chen Wei,Chen Huijiang,Wu Guanxiong,Tan Mingxiong,Liang Jintao,Zhou Zhide
An electrochemical aptasensor for quantitatively detecting glypican-3 (GPC3) was constructed by combining hemin-reduced graphene oxide-platinum (H-rGO-Pt) nanoparticles (NPs) with reduced graphene oxide-gold (rGO-Au) nanoparticles (NPs). Herein, the rGO-Au NPs deposited onto screen-printed
electrodes resulted in signal amplification due to their large surface areas. Meanwhile, highly conductive H-rGO-Pt NPs acted as a sensing medium that improved electrical conductivity and as an indicator for monitoring peak current for determination. A GPC3 aptamer (GPC3apt) with a low equilibrium
dissociation constant was used as a bio-recognition molecule. GPC3apt specifically captured GPC3 proteins and formed aptamer-GPC3 complexes, which impeded electron transfer and thus hampered the redox signal of hemin in H-rGO-Pt NPs. This developed electrochemical aptasensor showed a linear
response to GPC3 (from 0.001 μg/mL to 10 μg/mL) and had a detection limit of 0.001 μg/mL. This work provides a low-cost and highly sensitive detection with and good recovery for GPC3 and holds great promise for the clinical diagnosis of hepatocellular carcinoma.
研究领域:
生物医学工程
·
肿瘤学
·
纳米技术
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高影响因子
TALANTA
·
2023
·
IF 6.1
Lidong Pang,Ling'e Wang,Yaqi Liang,Zhenghui Wang,Wei Zhang,Qianyu Zhao,Xinyan Yang,Yujun Jiang
Escherichia coli O157: H7 ( E. coli O157: H7) is one of the most common foodborne pathogens and is widespread in food and the environment. Thus, it is significant for rapidly detecting E. coli O157: H7. In this study, a colorimetric aptasensor based on aptamer-functionalized magnetic beads, exonuclease III (Exo III), and G-triplex/hemin was proposed for the detection of E. coli O157: H7. The functional hairpin HP was designed in the system, which includes two parts of a stem containing the G-triplex sequence and a tail complementary to cDNA. E. coli O157: H7 competed to bind the aptamer (Apt) in the Apt-cDNA complex to obtain cDNA. The cDNA then bound to the tail of HP to trigger Exo III digestion and release the single-stranded DNA containing the G-triplex sequence. G-triplex/hemin DNAzyme could catalyze TMB to produce visible color changes and detectable absorbance signals in the presence of H 2 O 2 . Based on the optimal conditions, E. coli O157: H7 could be detected down to 1.3 × 10 3 CFU/mL, with a wide linear range from 1.3 × 10 3 to 1.3 × 10 7 CFU/mL. This method had a distinguished ability to non-target bacteria, which showed good specificity. In addition, the system was successfully applied to detect E. coli O157: H7 in milk samples.
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MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 2
·1-8
·
IF 6.232
Li Ruiying,Liu Qiang,Jin Yan,Li Baoxin
The authors have incidentally found that the three tandem repeats of a 13-mer G-rich oligomer (with sequence 5′-TGG GAA GGG AGG G-3′; referred to as G3) can directly fold into a stable G3 trimer. The G3 trimer/hemin DNAzyme exhibits an about 3-fold higher peroxidase-mimicking activity compared to the conventional G3/hemin DNAzyme. Combining this finding with rolling circle amplification (RCA), a colorimetric assay was developed for sensitive and specific determination of microRNA. In this method, each cycle of RCA generates three catalytic units. This leads to a significant signal amplification of the RCA. Using let-7a as a model analyte, the colorimetric method (best performed at 420 nm) exhibits high sensitivity toward microRNA-let-7a with a 37 fM detection limit and an analytical range that covers 3 orders of magnitude. The method was applied to the determination of let-7a in some cell lysates.
This G-triplex trimer-based rolling circle amplification (RCA) method can produce three catalytic units per RCA cycle, which can significantly improve the amplification efficiency of RCA.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2015
·
IF 4.097
Yuezhen He,Jian Sun,Xiaoxun Wang,Lun Wang
A sandwich chemiluminescence (CL) immunosensor for sensitive detection of human leptin was developed with a multiple signal amplification strategy from catalytic hemin/G-quadruplex DNAzymes and functional superparamagnetic nanocomposites. To construct this sensing platform, core–shell structural Fe 3 O 4 /polydopamine (PD)/Au superparamagnetic nanocomposites were synthesized by an in situ method with PD as a substrate adhesive. These nanocomposites were further characterized by scanning electron microscopy, transmission electronic microscopy, energy-dispersive spectroscopy, Fourier transform infrared spectrum and a vibrating sample magnetometer at room temperature. In this immunosensor, the monoclonal anti-human leptin antibody (capture antibody) was bound to the Fe 3 O 4 /PD/Au nanocomposites. Human leptin, biotinylated detection antibodies and streptavidin-DNAzymes were successively combined the above-mentioned nanocomposites to form sandwich-type immunocomplex through specific interactions. The magnetic particles loaded with the immunocomplex were separated by an external magnet, and the DNAzymes in the immunocomplex greatly enhanced the CL emission of the luminol–H 2 O 2 system. The immunosensor exhibited a high sensitivity, a good specificity, and a wide linear range for human leptin detection from 1.0 pg mL −1 to 8.0 × 10 2 pg mL −1 with a low detection limit of 0.3 pg mL −1 . This sensor is one of the most sensitive methods for leptin detection due to the highly efficient catalysis of the DNAzymes and analyte enrichment on magnetic capture.
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CHEMISTRY-A EUROPEAN JOURNAL
·
2016
·Vol. 22
·期 50
·18001-18008
·
IF 5.771
Jing Liu,Meirong Cui,Li Niu,Hong Zhou,Shusheng Zhang
Graphical Time for gold : A smart electrochemical aptamer biosensor for K562 leukemia cancer cells with enhanced peroxidase-like properties that shows a good performance in selectivity and sensitivity is presented. The biosensor employs a new graphene-family ternary composite with prickly Au nanoflowers on its surface. Graphene composites with hemin and gold nanoparticles show a better performance for hydrogen peroxide decomposition compared to that of the three components alone or duplex/hybrid complexes. Our previous studies showed that the morphology of the Au nanoparticles may greatly influence the catalytic activity of graphene-family peroxidase mimics. Recently, we found that Au nanoflowers could grow in situ and form on the surface of hemin/RGO (reduced graphene oxide). The prickly morphology of this Au nanoflower brought a higher catalytic ability with enhanced kinetic parameters than traditional Au nanoparticles that showed a smooth surface. Therefore, based on this discovery, a smart electrochemical aptamer biosensor for K562 leukemia cancer cells was further presented with good performance in selectivity and sensitivity attributed to the excellent mimetic peroxidase catalytic activity of this newly synthesized Au nanoflower decorated graphene–hemin composite (H-RGO-Au NFs).
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NEW JOURNAL OF CHEMISTRY
·
2023
·Vol. 47
·期 47
·21654-21660
·
IF 3.3
Ye He,Panlin Wang,Zhuzheng Wu,Yating Chen,Xiaohao Gan,Fangjie Li,Wenxiang Wang
The precise identification and detection of circulating tumor cells (CTCs) is significant for the early diagnosis and prognostic treatment of cancer. However, it remains a challenge to detect CTCs with high sensitivity and specificity. In this study, a colorimetric sensor for the analysis of CTCs was constructed based on dual-aptamer recognition and core (Fe3O4)–shell (Pt) nanoparticle (Fe3O4@Pt NP) nanozymes coupled with a G-quadruplex as a signal transducer/amplifier. The results show that the linear range of the designed colorimetric sensor is 50–4000 cells and the detection limit is 4 cells. The constructed aptasensor could be extended to identify various CTCs by changing the aptamers, and holds great potential in precise tumor diagnosis.
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BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Xingxing Zhou,Shijing Guo,Jiaxi Gao,Jianmin Zhao,Shuyan Xue,Wenju Xu
Based on cascade catalysis amplification driven by glucose oxidase (GOx), a sensitive electrochemical impedimetric aptasensor for protein (carcinoembryonic antigen, CEA as tested model) was proposed by using Cu-based metal-organic frameworks functionalized with Pt nanoparticles , aptamer , hemin and GOx ( [email protected] ). CEA aptamer loaded onto [email protected] was bound with hemin to form [email protected] (hGq) with mimicking peroxidase activity. Through sandwich-type reaction of target CEA and CEA aptamers (Apt1 and Apt2), the obtained [email protected] as signal transduction probes (STPs) was captured to the modified electrode interface. When 3,3-diaminobenzidine (DAB) and glucose were introduced, the cascade reaction was initiated by GOx to catalyze the oxidation of glucose, in situ generating H 2 O 2 . Simultaneously, the decomposition of the generated H 2 O 2 was greatly promoted by [email protected] and hGq as synergistic peroxide catalysts, accompanying with the significant oxidation process of DAB and the formation of nonconductive insoluble precipitates (IPs). As a result, the electron transfer in the resultant sensing interface was effectively hindered and the electrochemical impedimetric signal (EIS) was efficiently amplified. Thus, the high sensitivity of the proposed CEA aptasensor was successfully improved with 0.023 pg mL −1 , which may be promising and potential in assaying certain clinical disease related to CEA.
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PATHOLOGY RESEARCH AND PRACTICE
·
2024
·
IF 2.9
Haiping Zhang,Wei Li
Background Intracerebral hemorrhage (ICH) is one deadly subtype of stroke with high morbidity and mortality. Oxidative stress and inflammation are major factors contributing to blood-brain barrier (BBB) dysfunction, which induces perihematoma edema and exacerbates ICH-induced secondary brain injury. SYN1 is a crucial neuronal phosphoprotein that plays a pivotal role in neuronal development. Our study was devised to clarify the function of SYN1 in hemin-induced cell injury in PC12 cells and brain injury in ICH rat models. Methods For in vitro assay, PC12 cells were first stimulated by 150 µM hemin for 4 h and then transfected with si-SYN1 and pcDNA3.1-SP1. Cell viability was tested through lactate dehydrogenase (LDH) release and CCK-8 assays. Cell apoptosis was observed through TUNEL staining. Inflammatory factors (IL-1β, IL-6, and TNF-α) were evaluated through western blotting. Oxidative markers (GSH, MDA, and SOD) were detected by adopting commercial kits. The binding sites of SP1 on the SYN1 promoter were predicted by using the JASPAR database and were validated through performing CHIP and luciferase reporter assays. SP1 and SYN1 expression in PC12 cells was investigated by RT-qPCR. For in vivo assay, rats were administrated intracerebroventricularly with si-SYN1 at 72 h before ICH induction. SYN1 and SP1 expression in ICH rats was assessed by RT-qPCR. Neurological deficits, brain edema, and BBB disruption at 24 h following ICH were assessed by conducting neurobehavioral tests, brain water content examination, and Evans blue extravasation assay. Results In vitro, hemin stimulation elevated SYN1 expression, attenuated cell viability, enhanced apoptosis, and induced inflammation and oxidative stress in PC12 cells, which were offset by SYN1 knockdown. SP1 bound to the SYN1 promoter region and positively regulated SYN1 expression. Overexpressing SP1 antagonized the impacts of SYN1 downregulation on hemin-induced damage in PC12 cells. In vivo , SYN1 and SP1 expression was upregulated in ICH rat models. Administration of si-SYN1 effectually improved neurological function, attenuated brain edema, and ameliorated BBB disruption following ICH. Conclusion SP1-mediated SYN1 probably participates in the process of neuronal damage, BBB disruption, and brain injury following ICH.
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MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 6
·1-12
·
IF 5.3
Chen Yafei,Zhang Xuhui,Huang Wanjin,Jin Zhenhuan,Wei Xiaoping,Li Jianping
Atrazine (ATZ) is used extensively, resulting in residues in food and the environment, posing a serious threat to human health. Herein, Cd 0.5 Zn 0.5 S/Ti 3 C 2 photoelectric material was synthesized and immobilized on a FTO electrode as a photoanode. A photoelectrochemical (PEC) aptamer sensor was constructed for the highly sensitive and selective determination of ATZ based on signal amplification via cascade catalysis of CRISPR/Cas12a and G-quadruplex/hemin DNAzyme (G4/hemin DNAzyme). G4/hemin DNAzyme catalyses the oxidation reaction between H 2 O 2 and dopamine (DA) to form polydopamine (PDA) deposit. This process, in turn, inhibits the photocurrent at the photoanode, leading to a decrease in photocurrent. Concurrently, the depletion of DA as an electron donor for the PEC reaction at the photoelectrode further contributes to the decrease in photocurrent. ATZ can hybridize with ATZ aptamer (Apt) in Apt/cDNA to release activation strand (cDNA), which activates the activity of CRISPR/Cas12a and triggers cleavage of G4, causing the cleaving of G4/hemin DNAzyme immobilized on the electrode surface. This process leads to a decrease of G4/hemin DNAzymes amount on the electrode, consequently reducing both the PDA generation and the DA consumption. As a result, the photocurrent is restored. The cascade catalysis of CRISPR/Cas12a and G4/hemin DNAzyme has been demonstrated to result in photocurrent amplification. The photocurrent change was linear with the logarithmic value of ATZ concentration in the range 1.00 × 10 –12 to 1.00 × 10 –5 mol/L. The limit of detection was 3.47 × 10 –13 mol/L. The sensor has been successfully applied to the determination of trace ATZ in environmental and food samples. Graphical abstract
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ANALYST
·
2017
·Vol. 142
·期 11
·2013-2019
·
IF 3.885
Hui Zhang,Zhihui Guo,Huilei Dong,Hongfei Chen,Chenxin Cai
In this work, a highly sensitive electrochemiluminescence (ECL) assay was fabricated for the detection of human DNA (cytosine-5)-methyltransferase1 (DNMT1) activity in cancer cells. The ECL assay coupled hybridization chain reaction with a G-quadruplex/hemin DNAzyme biosensing strategy. The ECL intensity changes (ΔI) allowed detection of DNMT1 activity down to 0.09 U mL−1, and ΔI was proportional to the logarithm of the activity of DNMT1 within the range of 1.0 to 30.0 U mL−1 in buffer solution. It also showed high sensitivity to DNMT1 activity in A549 cells, with a detection limit of about 2 cells. This ECL assay provides a promising platform for profiling of the mutational cells of tumors and shows a great potential for application to DNA methylation-related clinical diagnostics.
研究领域:
生物化学
·
分子生物学
·
临床诊断学
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SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Nuanfei Zhu,Xuesong Li,Ye Liu,Jingfu Liu,Yawei Wang,Xiangyang Wu,Zhen Zhang
Dibutyl phthalate (DBP) is considered as one of the most widely used phthalate esters (PAEs), which has attracted worldwide concerns because of its potential threats to eco-environments and human health. Systematic investigations of DBP environmental occurrence contribute to the further risk assessment, which depends on effective and available analytical methods. In this study, an amplified ratiometric fluorescence ELISA was established for sensitive and high-throughput detection of DBP in the aquatic system based on a novel tetrahedral DNA nanostructure (TDN)-scaffolded-DNAzyme (Tetrazyme). Wherein, Tetrazyme was prepared by the precise folding of G-quadruplex sequence on three vertex angles of the TDN, together with hemin as the horseradish peroxidase (HRP)-mimicking enzyme. The rigid TDN avoided the local overcrowding effect to provide a reasonable spatial spacing on the interface for G-quadruplex sequence, increasing the collision chance between DNAzyme and substrates, improving the catalytic ability of DNAzyme effectively. Besides, streptavidin (SA) and biotin (bio) were used to anchor TDN and antibody, in which the specific binding of SA/bio could make more Tetrazyme conjugate on each signal element, resulting in the dual signal amplification. Meanwhile, the accuracy and precision were enhanced owing to the inherent built-in rectification to the environment from the dual output ratiometric fluorescence assay. Under the optimized conditions, the detection limit of this proposed method was 0.17 ng/mL (16 times lower than that of conventional ELISA using the same antibody) with a satisfactory accuracy (recoveries, 79.0%- 116.2%; CV, 2.1–6.5%). Overall, this platform provides a promising way for accurate, sensitive and rapid determination of DBP from environmental waters.
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Food Analytical Methods
·
2014
·Vol. 8
·期 6
·1556-1564
·
IF 1.802
Liu Tao,Zhang Xiao,Hao Jianpeng,Zhu Wenxin,Liu Wei,Zhang Daohong,Wang Jianlong
Herein, we for the first time report an acetylcholinesterase (AChE)-free colorimetric method for the detection of chlorpyrifos (CP), which is based on the oxidation reaction of H 2 O 2 with CP and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS 2− ) catalyzed by hemin/G-quadruplex DNAzyme. In the absence of CP, ABTS 2− is directly oxidized by H 2 O 2 with hemin/G-quadruplex DNAzyme as a biocatalyst to produce the blue-green-colored free radical anion (ABTS ·− ), accompanied with an obvious UV-visible absorbance at 418 nm. However, the amount of H 2 O 2 used in H 2 O 2 -DNAzyme-ABTS 2− system will be decreased if CP is first reacted with H 2 O 2 , leading to the decrease of the amount of ABTS ·− and a change in the UV-visible absorbance of ABTS ·− at 418 nm. Thus, an indirect dependence relation between CP concentration and the absorbance of ABTS ·− at 418 nm could be constructed and used for CP detection. Under optimal conditions, the detection range of the method is 0.04–1 μg/mL, with a detection limit of 0.01 μg/mL. Application of the method in real juice samples shows good analytical features in terms of recoveries (95–107.5 %), the relative standard deviation (RSD) (0.6–4.7 %), rendering it as an attractive candidate to current methodologies for the determination of CP.
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危险品化学品经营许可证（带存储）