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FOOD CHEMISTRY · 2021 · IF 7.514

A turn-on fluorescence sensor based on Cu2+ modulated DNA-templated silver nanoclusters for glyphosate detection and mechanism investigation

Yixia Yang,Behafarid Ghalandari,Liyun Lin,Xiao Sang,Wenqiong Su,Adeleh Divsalar,Xianting Ding

The abuse application of glyphosate can result in a potential hazard for environment and human, however its ultrasensitive detection remains challenging. Herein, a Cu 2+ modulated DNA-templated silver nanoclusters (DNA-AgNCs) sensor was constructed to sensitively determine glyphosate based on the turn-on fluorescence strategy. The fluorescence quenching of DNA-AgNCs occurred with the existence of Cu 2+ . Upon the presence of glyphosate, the functional groups on the surface of glyphosate could chelate with Cu 2+ , following the fluorescence recovery of DNA-AgNCs. Through the stoichiometric methods, we unveil that Cu 2+ -trigged fluorescence quenching mode is a combination of static and dynamic quenching with the static mode being predominant. In DNA-AgNCs/Cu 2+ system, the carboxylate, amine, and phosphonate groups of glyphosate interact with Cu 2+ through chelation, in which the carboxylate oxygen, the phosphonate oxygen atoms, and the monoprotonated secondary amine nitrogen atom and Cu 2+ form chelate rings. This fluorescence sensor showed a desired linearity of glyphosate analysis under the optimum conditions, ranging from 15 to 100 μg/L with a low detection down to 5 μg/L. Moreover, the proposed sensor was successfully utilized to measure glyphosate in real samples, indicating a promising application in pesticide residues detection.

CAS / MDL: 10605-21-7 23564-05-8
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ANALYTICAL CHEMISTRY · 2021 · IF 6.986

Hybridization Chain Reaction-Amplified Electrochemical DNA-Based Sensors Enable Calibration-Free Measurements of Nucleic Acids Directly in Whole Blood

Shaoguang Li,Hongxing Li,Xun Li,Man Zhu,Hui Li,Fan Xia

Hybridization chain reaction (HCR) amplification strategy has been extensively explored for the application of electrochemical DNA-based sensors. Despite the enhancement in its sensitivity using the HCR, such sensor platform exhibited significant sensor-to-sensor variations in current due to variations in probe counts and lengths. To circumvent this, we are developing here a calibration-free “O-N” approach to generate a ratiometric, unitless value that is independent of these variations. Specifically, this approach employs two types of redox reporters, denoted as “One reporter” and “N reporters”, with the former attached on the capture DNA and the latter on H1 and H2 strands. By optimizing the attachment sites of these reporters onto DNA strands, we demonstrate a significantly enhanced sensitivity of such sensor platform by four orders of magnitude, achieving accurate, calibration-free measurement of nucleic acids including ctDNA directly in undiluted whole blood without the requirement to calibrate each individual sensor.

CAS / MDL: 1633-78-9
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MATERIALS & DESIGN · 2021 · IF 7.991

DNA directed immobilization of horseradish peroxidase on phase- transitioned lysozyme modified TiO2 for efficient degradation of phenol in wastewater

Baolin Chen,Xun Wang,Xia Gao,Jie Jiang,Mancheng Hu,Shuni Li,Quanguo Zhai,Yucheng Jiang

In this work, TiO 2 nano particle was used for two purposes: it played as a role of a solid carrier for immobilization of HRP, and meanwhile acted as a photo-catalyst itself. In this way, the loss of enzymatic activity during enzyme immobilization can be compensated by the coupling of TiO 2 photo-catalysis. Then, the surface of TiO 2 nanoparticles were modified by phase-transitioned lysozyme (PTL) and connected with horseradish peroxidase (HRP) via a DNA helix ( [email protected] 2 ). The DNA double helix chain was utilized as a flexible spacer arm to keep the flexibility of the immobilized enzyme. The [email protected] 2 showed a high recovered activity (90.52%) of the original activity of free HRP. This data is better than most reported activity recovery of immobilized enzyme. Moreover, [email protected] 2 can maintain high catalytic activity, stability and reusability when degrading phenol in wastewater. The phenol of 0.5 mmol∙L −1 could be totally degraded within 10 min. Moreover, a visual colorimetric method for phenol concentration by [email protected] 2 was established, and the phenol concentration of 0–0.2 mmol∙L −1 can be visually detected with naked eyes. Furthermore, an enzymatic-photocatalytic synergetic effect of [email protected] 2 is found and investigated. The efficiency of [email protected] 2 ensures it a potential application in the wastewater treatment.

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RSC Advances · 2020 ·Vol. 10 ·期 58 ·35374-35380 · IF 3.119

An aptasensor for the label-free detection of thrombin based on turn-on fluorescent DNA-templated Cu/Ag nanoclusters

Baozhu Zhang,Chunying Wei

A highly sensitive thrombin aptasensor was constructed based on the alteration of the aptamer conformation induced by the target recognition and the turn-on fluorescence due to the proximity of two darkish DNA-templated copper/silver nanoclusters (DNA-Cu/Ag NCs). Two DNA templates were designed as the functional structures consisting of the Cu/Ag NC-nucleation segment located at two termini or one terminus and the aptamer segment in the middle of a DNA template. Two darkish DNA-Cu/Ag NCs came close to each other when the aptamer combined with the target due to the conformational alteration of the aptamer structure, resulting in an increased fluorescence signal readout. Thrombin was sensitively determined as low as 1.6 nM in the range of 1.6–8.0 nM with a high selectivity. Finally, this sensor succeeded in detecting thrombin in a real fetal bovine serum.

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ACS Applied Materials & Interfaces · 2020 · IF 8.758

Novel Protease-Free Long-Lasting Chemiluminescence System Based on the Dox-ABEI Chimeric Magnetic DNA Hydrogel for Ultrasensitive Immunoassay

Haiping Wu,Min Zhao,Jia Li,Xiaoyan Zhou,Tiantian Yang,Dezhang Zhao,Ping Liu,Huangxian Ju,Wei Cheng,Shijia Ding

Most of chemiluminescence (CL) substrates exhibit the flash-type light emission. Therefore, the long-lasting CL system is always the crown in the field of CL-based analysis methodology. In this work, we constructed a Dox-ABEI chimeric magnetic DNA hydrogel (MDH) as a novel protease-free long-lasting CL reaction system. The functional MDH can transform flash-type ABEI/H2O2/CO2+ reaction into a glow-type CL system because of its block effect on delaying the diffusion rate of co-reactants, making the CL reaction gradually occur. More importantly, the functional MDH possessed the advantages of biocompatibility and controllability and could be well-designed to incorporate different biosensing strategies. Subsequently, we established a functional MDH-based long-lasting CL immunoassay system for ultrasensitive and highly specific detection of d-dimer and fibrin degradation products (FDPs). The designed CL immunoassay can detect d-dimer and FDP down to 53.7 and 31.6 fg/mL, respectively, with a wide line ranging from 100 fg/mL to 100 ng/mL, which was superior to the previously reported CL biosensing strategies. Moreover, benefiting from the magnetic separation of MDH and excellent CL performance, the developed immunoassaying method was successfully applied in the detection of clinical samples, which showed a close correlation with clinical reference technology. Thus, this functional MDH proved to be an excellent long-lasting CL system and a potential technical platform for clinical bioanalysis applications.

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DIGESTIVE DISEASES AND SCIENCES · 2020 ·Vol. 66 ·期 10 ·3375-3390 · IF 2.751

DNA Damage-Regulated Autophagy Modulator 1 (DRAM1) Mediates Autophagy and Apoptosis of Intestinal Epithelial Cells in Inflammatory Bowel Disease

Zhang Yu,Li Xiaozhi,Li Yaoting,Li Ying,Wang Ying,Zhu Liguo,Chen Peng,Tian Zhenyi,Qiu Yun,Feng Rui,Zeng Zhirong,He Yao,Chen Baili,Zhang Shenghong,Mao Ren,Chen Minhu

Background and Aims DNA damage-regulated autophagy modulator 1 (DRAM1) is required for induction of autophagy and apoptosis. However, the influence of DRAM1 on the pathogenesis of inflammatory bowel disease (IBD) has not been explored. Methods DRAM1 expression was examined in the intestinal mucosa of patients with IBD and colons of colitis mice. We used a recombinant adeno-associated virus carrying small hairpain DRAM1 to knock down the DRAM1 gene to treat colitis in the mice. The effect of DRAM1 on autophagy and apoptosis of intestinal epithelial cells was explored. DRAM1-mediated interaction with the c-Jun N-terminal kinase (JNK) pathway was also examined. Results DRAM1 expression in the intestinal mucosa of the IBD patients was higher than that in the control participates. DRAM1 expression in the inflammatory cells in patients with Crohn’s disease (CD) was lower than that in patients with ulcerative colitis (UC). Additionally, DRAM1 expression was correlated with the Simple Endoscopic Score for CD and the Mayo endoscopic score for UC. Serum levels of DRAM1 in the IBD group were substantially higher than those in the normal group. The knockdown of DRAM1 could alleviate colitis symptoms in mice. In in vitro experiments, knocking down DRAM1 could reduce autophagy and apoptosis levels. Mechanistically, DRAM1 may participate in the regulation of these two processes by positively regulating JNK activation. Conclusions During intestinal inflammation, the upregulation of DRAM1 may promote the activation of JNK and further aggravate intestinal epithelium damage.

CAS / MDL: 9005-64-5
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ACS Applied Bio Materials · 2020 · IF 0

Sensitive Fluorescence Aptasensor Based on Hybridization Chain Reaction with Upconversion Nanoparticles by Triplex DNA Formation for Bisphenol A Detection

Yu Wang,Xudong Zhao,Bingyang Huo,Shuyue Ren,Jialei Bai,Yuan Peng,Shuang Li,Dianpeng Han,Jiang Wang,Tie Han,Zhixian Gao

Application of nanotechnology to detection can effectively increase the biocompatibility of detection sensors. However, since the traditional signal output element has a high background value, the detection sensitivity is often limited. Here, we designed a fluorescence aptasensor based on hybridization chain reaction (HCR) and triplex structure (TS). The aptamer was competitively bound by bisphenol A (BPA) and complementary DNA (cDNA). Unbound cDNA was obtained by magnetic separation. HCR could be triggered by cDNA to form long double-stranded DNA (dsDNA) products. The upconversion nanoparticle (UCNP)-modified ssDNA can form a TS with the HCR product. Thereby, the distance between the UCNPs and Dabcyl on the hairpin was brought closer, and fluorescence was quenched to generate a signal change. HCR and the low background value of UCNPs could improve the sensitivity of detection. The aptasensor has a detection limit as low as 0.057 ng/mL and has been successfully applied to actual sample detection. Its high sensitivity, strong specificity, and excellent biocompatibility make it promising for other target detection and clinical diagnosis.

CAS / MDL: 111-30-8 71-44-3
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BIOSENSORS & BIOELECTRONICS · 2021 · IF 10.618

Entropy-driven amplified electrochemiluminescence biosensor for RdRp gene of SARS-CoV-2 detection with self-assembled DNA tetrahedron scaffolds

Zhenqiang Fan,Bo Yao,Yuedi Ding,Jing Zhao,Minhao Xie,Kai Zhang

Dependable, specific and rapid diagnostic methods for severe acute respiratory syndrome β -coronavirus (SARS-CoV-2) detection are needed to promote public health interventions for coronavirus disease 2019 (COVID-19). Herein, we have established an entropy-driven amplified electrochemiluminescence (ECL) strategy to detect the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2 known as RdRp-COVID which as the target for SARS-CoV-2 plays an essential role in the diagnosis of COVID-19. For the construction of the sensors, DNA tetrahedron (DT) is modified on the surface of the electrode to furnish robust and programmable scaffolds materials, upon which target DNA-participated entropy-driven amplified reaction is efficiently conducted to link the Ru (bpy) 3 2+ modified S3 to the linear ssDNA at the vertex of the tetrahedron and eventually present an “ECL on” state. The rigid tetrahedral structure of the DT probe enhances the ECL intensity and avoids the cross-reactivity between single-stranded DNA, thus increasing the sensitivity of the assays. The enzyme-free entropy-driven reaction prevents the use of expensive enzyme reagents and facilitates the realization of large-scale screening of SARS-CoV-2 patients. Our DT-based ECL sensor has demonstrated significant specificity and high sensitivity for SARS-CoV-2 with a limit of detection (LOD) down to 2.67 fM. Additionally, our operational method has achieved the detection of RdRp-COVID in human serum samples, which supplies a reliable and feasible sensing platform for the clinical bioanalysis.

CAS / MDL: 25952-53-8 6066-82-6
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SENSORS AND ACTUATORS B-CHEMICAL · 2020 · IF 7.1

A DNA structure-mediated fluorescent biosensor for apurinic/apyrimidinic endonuclease 1 activity detection with ultra-high sensitivity and selectivity

Yuqiang Hu,Zhen Zhang,Weicong Ye,Wei Zhang,Minghao Hu,Wenqian Yuan,Hongbo Wang,Xianjin Xiao,Tongbo Wu

Human apurinic/apyrimidinic endonuclease 1 (APE1) plays a crucial role in DNA repair and gene regulation. The abnormal variations of the concentration of APE1 in the human blood and tissue cells are highly correlated to various diseases. Thus, APE1 can be used as a biomarker to aid clinical diagnosis of diseases. Detection of APE1 activity with high sensitivity and selectivity simultaneously is difficult to achieve. Here we provide a DNA structure-mediated fluorescent biosensor to solve the above problem. Upon the existence of APE1, the apurinic/apyrimidinic site will be cleaved, and the terminal structure of the hairpin DNA substrate will change. Then the terminal deoxynucleotidyl transferase and endonuclease IV could provide dual fluorescent signal amplification. Through the ingenious structure design of the biosensor, we have improved the reactivity of APE1 and minimized the interference of other enzymes. The results indicate that the detection limit for APE1 is as low as 1.7 × 10 −6 U/mL. The biosensor has been successfully applied to the detection of APE1 in real biological samples and the screening of APE1 inhibitors.

CAS / MDL: 6960-45-8
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Nanoscale · 2020 ·Vol. 12 ·期 44 ·22574-22585 · IF 6.895

Isolation of DNA aptamers targeting N-cadherin and high-efficiency capture of circulating tumor cells by using dual aptamers

Tian Gao,Pi Ding,Wenjing Li,Zhili Wang,Qiao Lin,Renjun Pei

Circulating tumor cells (CTCs) acquire mesenchymal markers (e.g., N-cadherin) and lose epithelial markers (e.g., epithelial cell adhesion molecule, EpCAM) during the epithelial-mesenchymal transition (EMT) and are therefore ideal biomarkers of tumor metastasis. However, it is still a challenge to efficiently capture and detect circulating tumor cells with different phenotypes simultaneously. In this work, to obtain aptamers targeting N-cadherin in the native conformation on live cells, we established stable N-cadherin overexpressing cells (N-cadherin cells) and used these cells to identify a panel of N-cadherin-specific aptamers through the cell-SELEX approach. Two aptamer candidates obtained after 12 rounds of selection showed a low equilibrium dissociation constant in the nanomolar range, indicating high binding affinity. The truncated aptamer candidate NC3S showed the highest binding affinity to N-cadherin cells with a low Kd value of 20.08 nM. The SYL3C aptamer was reported to target cancer cell surface biomarker EpCAM. Then, we synthesized two kinds of aptamer-modified magnetic nanoparticles (SYL3C-MNPs and NC3S-MNPs). Both SYL3C and NC3S aptamers possess excellent capture specificity and efficiency for the target cells. The aptamer–MNP cocktail exhibits a considerable capture efficiency and sensitivity for rare cancer cells of epithelial and mesenchymal phenotypes. Furthermore, no CTCs were found in blood samples from healthy donors, while CTCs were successfully isolated by using the aptamer–MNP cocktail for 15 out of 16 samples collected from patients. In summary, the two kinds of aptamer-modified MNPs could be utilized as a promising tool for capturing CTCs from clinical samples.

CAS / MDL: 25322-68-3
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ANALYST · 2021 ·Vol. 146 ·期 7 ·2321-2329 · IF 4.616

Highly sensitive detection of DNA damage in living cells by SERS and electrochemical measurements using a flexible gold nanoelectrode

Jing Zhou,Dan Yang,Guohui Liu,Siying Li,Wennan Feng,Guocheng Yang,Jin He,Yuping Shan

Guanine (G) oxidation products, such as 8-hydroxy-2′-deoxyguanosine (8-OHdG) and 8-oxo-guanine (8-OXOG), have been widely studied as promising biomarkers for DNA oxidative damage. In this work, we develop a new method to detect G oxidative products released from live cells after chromium (VI) ion or hydrogen peroxide treatments by using a glass nanopipette-based flexible gold nanoelectrode (fGNE). Specific response to G oxidative products with high sensitivity can be detected from the fGNE tip through integrated electrochemical measurements and surface-enhanced Raman spectroscopy. The fGNE apex can be positioned very close to the cell membrane noninvasively because of its high flexibility and nanoscale tip size. With the assistance of the electrophoretic force, the fGNEs can effectively collect and detect the G-derived DNA damage products released from individual cells in the cell culture medium with high sensitivity.

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ANALYTICA CHIMICA ACTA · 2020 · IF 5.977

Catalytic hairpin assembly-triggered DNA walker for electrochemical sensing of tumor exosomes sensitized with Ag@C core-shell nanocomposites

Yuehua Guo,Qihang Cao,Qiumei Feng

The detection of a small number of exosomes provides the possibility for early cancer diagnosis and prognosis. Here, a multi-signal amplified electrochemical sensing platform was explored for the ultrasensitive detection of tumor exosomes relying on catalytic hairpin assembly-triggered DNA walker, entropy beacon-based DNA assembly and Ag@C core-shell nanocomposites. In this work, the utilization of Ag@C nanocomposites as electrode interface effectively enhanced functional active sites and electron transfer capability. By designing a target-assisted entropy beacon-based DNA assembly, single exosome initiated the release of multiple special DNA sequences , which could be separated conveniently by magnet and then hybridize with the blocking DNA to liberate swing arm. DNA walker was activated with the assistance of catalytic hairpin assembly, introducing extensive electroactive methylene blue (MB) to electrode surface. Thus, the detection of exosomes was transferred into the measurement of the MB current, with a good liner range from 100 to 75 000 particles/μL. Furthermore, this constructed sensing system displayed acceptable reproducibility, long-term stability, favorable selectivity , and highlighting application potential in real samples.

CAS / MDL: 9012-76-4
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