JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2020
·
IF 3.807
Jingxiu Mao,Xin Chen,Huahuang Xu,Xueqin Xu
T4 polynucleotide kinase (T4 PNK), a bifunctional repair enzyme, catalyzes the transfer of γ-phosphate residues from adenosine triphosphate (ATP) to 5′- hydroxyl termini in nucleic acids，which plays an essential role in regulating DNA phosphorylation modes during the repair of damaged DNA. Herein, a sensitive DNAzyme-driven DNA walker biosensor was developed for amplified electrochemical detection of T4 PNK activity and inhibition based on T4 PNK-triggered phosphorylation reaction and λ exonuclease (λ exo) cleavage. A novel functional hairpin-shaped swing arm DNA (SA) containing a DNA sequence of the Pb 2+ -dependent DNAzyme is designed to specifically bind to and hydrolyze the substrate strand (rSS) sequence that can drive DNA walker. Upon the reaction of SA， T4 PNK and ATP, λ exo removes the mononucleotides from the stem of the phosphorylated SA, unfolding the hairpin structure and initiating DNAzyme-driven DNA walker. Subsequently, the DNA walker can repeatedly bind and cleave rSS with the aid of Pb 2+ , releasing the methylene blue (MB) - labeled fragment of rSS. The T4 PNK activity can be facilely monitored by recording the electrochemical signal change of MB. The proposed biosensor exhibited a broad linear range from 0.01 U mL −1 to 15 U mL −1 with a low detection limit of 0.001 U mL −1 . The proposed biosensor has also been utilized to screen the T4 PNK inhibitors and exhibited good performance in real sample analysis, making such a strategy hold potential promise in biological fields and clinical diagnosis.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
临床诊断
✓ 已验证
产品关联
DOI
高影响因子
BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Xianhua Shi,Yixuan Xie,Linxi Chen,Jing Lu,Luyong Zhang,Duanping Sun
Zeolitic imidazolate frameworks (ZIFs), a famous subfamily of metal-organic frameworks (MOFs), are considered promising electrocatalysts. Herein, ZIF-67 was selected as an electrocatalyst for designing electrochemical sensors due to having the best electrocatalytic activity in ZIFs. To overcome the insufficient electrocatalytic activity of ZIFs, ZIF-67 derivatives (QZIF-67-X, where X represents calcination time) were obtained by calcining at 250 °C for a certain time. The porous structure of the precursor in QZIF-67-X is maintained, exposing more active centers. QZIF-67-X could accelerate electron transfer and lead to improve the electrocatalytic performance. Moreover, QZIF-67-2 was chosen as an Au nanoparticle-supported nanocarrier to further bind G-quadruplex/hemin DNAzymes with strong catalytic activity due to the best supporting activity of QZIF-67-2 among QZIF-67-X. The synergistic catalysis of QZIF-67-2 and G-quadruplex/hemin DNAzymes effectively amplified the reduction current signal of H 2 O 2 . The linear range of the prepared electrochemical sensor was 2 μM-65 mM, and the detection limit was 1.2 μM. Moreover, the real-time detection of H 2 O 2 from HepG2 cells was achieved by the sensor, providing a novel technique for efficient anticancer drug evaluation. These results suggested that QZIF-67 can be utilized as an efficient electrocatalyst for improving the sensitivity of sensors.
研究领域:
生物化学
·
医学检测
·
材料科学
✓ 已验证
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2014
·
IF 5.9
Hui Xia,Lingling Li,Zhouyang Yin,Xiandeng Hou,Jun-Jie Zhu
A dual signal amplification strategy for electrochemiluminescence (ECL) aptasensor was designed based on biobar-coded gold nanoparticles (Au NPs) and DNAzyme. CdSeTe@ZnS quantum dots (QDs) were chosen as the ECL signal probes. To verify the proposed ultrasensitive ECL aptasensor for biomolecules, we detected thrombin (Tb) as a proof-of-principle analyte. The hairpin DNA designed for the recognition of protein consists of two parts: the sequences of catalytical 8–17 DNAzyme and thrombin aptamer. Only in the presence of thrombin could the hairpin DNA be opened, followed by a recycling cleavage of excess substrates by catalytic core of the DNAzyme to induce the first-step amplification. One part of the fragments was captured to open the capture DNA modified on the Au electrode, which further connected with the prepared biobar-coded Au NPs-CdSeTe@ZnS QDs to get the final dual-amplified ECL signal. The limit of detection for Tb was 0.28 fM with excellent selectivity, and this proposed method possessed good performance in real sample analysis. This design introduces the new concept of dual-signal amplification by a biobar-coded system and DNAzyme recycling into ECL determination, and it is promising to be extended to provide a highly sensitive platform for various target biomolecules.
研究领域:
生物分析学
·
纳米技术
·
生物传感器
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 145
·期 1
·107-114
·
IF 4.019
Yu Zhou,Huiying Xu,Hua Wang,Bang-Ce Ye
Exosomes are membrane-enclosed phospholipid extracellular vesicles with a variety of tumor antigens which can be applied in the diagnosis and treatment of cancer due to the high secretion on the surface of cancer cells. Until now, many research studies on exosomes have been reported, but convenient and low-cost detection methods still need to be developed. Recently, we have developed a sensitive, simple and low-cost colorimetric aptasensor by designing a hairpin-like structure, which combined the highly specific MUC1 aptamer with a hemin/G-quadruplex for the detection of breast cancer exosomes. The hemin/G-quadruplex toward H2O2 reduction was used to generate an evidently strong colorimetric response owing to acting as a HRP-mimicking DNAzyme. The aptasensor is regarded as an “on–off” type switch, which strictly controls the process of reaction responding to the existence or not of exosomes. In our study, associated exosome detection limits are 3.94 × 105 particles per mL, which showed a higher sensitivity compared to commercial ELISA. Our method not only exhibited the advantages of being convenient and time-saving, with few instruments used, but also the signal generated using this method can be easily observed by the naked eye. Furthermore, the proposed strategy can well differentiate breast cancer patients from healthy individuals, demonstrating potential application in the analysis of clinical specimens.
研究领域:
癌症生物学
·
分析化学
·
生物技术
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Wenshan Li,Qingqing Li,Huaiyue Xia,Wenjie Liu,Guoxing Jing,Jianna Yu,Wen Liu,Mengtong Lin,Yujuan Huang
Ochratoxin A (OTA) is a compound of concern due to its potential health effects on humans. Detecting OTA in food is crucial for safeguarding public health. In this study, we fabricated a multi-DNAzyme cascade reaction-mediated colorimetric aptasensors for OTA detection, integrating autocatalytic Mg 2+ -dependent DNAzyme cleavage (MNAzyme) and an entropy-driven circuit. In brief, the recognition between the aptamer and target OTA led to the release of DNA1. Subsequently, DNA1 hybridized with DNA2, generating an upstream MNAzyme that facilitated the production of a downstream MNAzyme. These MNAzymes possess similar substrate binding arms, enabling them to catalyze the same substrate. The catalytic efficiency of MNAzymes towards the substrates was enhanced due to the increasing concentration of MNAzymes. The cleavage products then triggered an entropy-driven cycle to generate a signal. Under optimal conditions, the sensing system exhibited low detection limits of 48.97 fM for OTA. Additionally, the proposed aptasensor was successfully applied to quantitatively analyze OTA in food samples. Thus, the multi-DNAzyme cascade reaction-mediated colorimetric aptasensors offer an adaptable platform for detecting traces of OTA contaminant in food.
研究领域:
食品安全检测
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2017
·
IF 4.162
Huiyan Xu,Qiwang Wu,Hong Shen
Single nucleotide polymorphisms (SNPs) are widely existed in human genome and associated with many diseases. Traditional PCR-based methods for SNP genotyping require protein enzyme, precise control of temperature and removal of resultant products, making the whole process labor intensive and time consuming. Although G-quadruplex DNAzyme-based assays provide many advantages over traditional approaches, the relatively low catalytic activity of DNAzyme becomes an unfavorable factor in its application process. Therefore, amplification of DNAzyme for further determination is of great desire in bioanalysis . In this work, we have developed an enzyme-free and non-label DNAzyme sensor for SNP genotyping based on target-catalyzed hairpin assembly (CHA) for DNAzyme amplification. The proposed sensor, carried out on microfluidic chemiluminescence (CL) assay, can sensitively discriminate rs242557 hotspot-SNP, the A/G single-nucleotide variation on human chromosome associated with Alzheimer's disease, with an absolute detection limit of 0.3 fmol.
研究领域:
生物分析学
·
分子诊断学
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2020
·
IF 10.257
Ziqi Zhou,Yangzi Zhang,Mingzhang Guo,Kunlun Huang,Wentao Xu
A relatively-simple and high-efficient fluorescent magnetic biosensor based on DNAzyme was established for the detection of E. coli O157:H7. In order to solve the problem of weak signal and low sensitivity in the detection of foodborne pathogenic bacteria , we ingeniously designed a fluorescent sensor based on triple signal amplification of magnetic beads, DNAzyme and photoluminescence . In the detection process, the E. coli -specific RNA-cleaving DNAzyme can specifically identify the target protein in crude intracellular mixture (CIM), which caused its conformation changes and induced rolling circle amplification (RCA) to the generation and luminescence of copper nanoclusters (CuNCs). This cascade amplification design can capture weak signals in the sample. The biosensor also indicated a good linear range from 10 CFU mL −1 to 1000 CFU mL −1 and obtained a limit of detection (LOD) of 1.57 CFU mL −1 , which showed a relatively high sensitivity compared with other studies. Furthermore, the biosensor displayed high-efficient detection capability in 1.5 h and good reproducibility (relative standard deviations < 2%). It has been proved that this sensor is feasible to the detection of E. coli O157:H7 in drinking water and apple juice. Moreover, we found that the final detection product can effectively wrap the magnetic beads and can be driven by the magnetic field. And this unexpected discovery will provide ideas for the development of biosensing robots.
研究领域:
生物化学
·
食品科学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Wang Ren,Ying Zhang,Wei Tao Huang,Nian Bing Li,Hong Qun Luo
This work reported a label-free colorimetric assay for sensitive detection of Hg 2+ based on Hg 2+ -triggered hairpin DNA probe (H-DNA) termini-binding and exonuclease Ш (Exo Ш)-assisted target recycling, as well as hemin/G-quadruplex (DNAzyme) signal amplification . The specific binding of free Hg 2+ with the thymine–thymine (T–T) mismatches termini of H-DNA could immediately trigger the Exo Ш digestion, and then set free G-quadruplex segments and Hg 2+ . The Exo Ш impellent recycling of ultratrace Hg 2+ produced numerous G-quadruplexes. The corresponding DNAzymes catalyzed efficiently the H 2 O 2 -mediated oxidation of the ABTS 2− to the colored product in the presence of hemin . Using the color change as the output signal, and the Exo Ш-aided Hg 2+ recycling and DNAzyme as the signal amplifier, the ultrasensitive assay system successfully achieved visual detection of Hg 2+ as low as 1.0 nM by the naked eye, and was suitable for field monitoring. The calibration curve was linear in the range of 50.0 pM to 20.0 nM for Hg 2+ ( R =0.9962) with a detection limit of 10.0 pM. Moreover, this proposed strategy showed excellent selectivity , portability and low-cost, and was successfully applied to colorimetric detection of Hg 2+ in laboratory tap water and Jialing river water samples.
研究领域:
分析化学
·
环境科学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Weipeng Liu,Cuifen Gan,Weidan Chang,Aori Qileng,Hongtao Lei,Yingju Liu
Recently, mimic enzymes have obtained particular interest by their high activity, stability, and biocompatibility. In this work, by coupling copper hydroxide nanozyme and G-quadruplex/hemin DNAzyme to form a double-integrated mimic enzyme, a visual, sensitive and selective immunosensor was established to detect microcystin-LR (MC-LR). In this immunoassay , the microplates were modified with core-shell silica/nickel silicate as the substrate to capture MC-LR antigens. Then, Cu(OH) 2 nanocages with fine regulation were used as the label to capture the secondary antibody for immunoreaction and the DNA primer for propagation, followed by using hybridization chain reaction to amplify the DNA primer, thus numerous DNAzymes (G-quadruplex/hemin) can be formed on the surface of Cu(OH) 2 nanocages with the aid of hemin . Such double-integrated mimic enzyme including Cu(OH) 2 nanozymes and DNAzymes showed excellent peroxidase activity for the chromogenic reaction of 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), which realized the visual detection of MC-LR in the range from 0.007 to 75 μg/L with the detection limit of 6 ng/mL, and thus provided the probability for the portable assessment of MC-LR in real sample.
研究领域:
生物化学
·
环境科学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Nanoscale
·
2025
·
IF 5.1
Beibei Zhang,Xinyi Wang,Min Pan,Xinhao Li,Hong Wang,Yingwei Zhang
In this study, a novel dual-switch fluorescent biosensor based on a bipedal DNAzyme walker was developed for the highly sensitive detection of the tumour biomarker miRNA-141. By exploiting the typically elevated expression levels of miRNA-141 and apurinic/apyrimidinic endonuclease 1 (APE1) in tumour cells, an endogenous dual-gating system was constructed. The presence of both targets was required to initiate an enzyme-free strand displacement reaction, enabling target recycling and the subsequent release of bipedal DNAzyme walker components (D1/D2), which were efficiently isolated using streptavidin-modified magnetic beads. In the presence of Mg2+, the activated DNAzyme walker specifically cleaved fluorophore-labelled substrate strands on gold nanoparticles (AuNPs). Through repeated“walking–cleavage”cycles, fluorescence signals were progressively restored and amplified. By integrating AuNPs as carriers and magnetic separation technology, a cascade dual-amplification system based on non-enzymatic strand displacement and bipedal DNAzyme walking was established, markedly enhancing the detection sensitivity. The biosensor exhibited dual-switch-specific activation, effectively suppressing interference from non-target RNAs and other nucleases, and maintained a low background signal. It demonstrated excellent discrimination between tumour cells and normal cells, and showed good anti-interference performance in spiked serum samples. This study provides a new strategy for the development of high-precision tumour biomarker detection, offering potential for trace analysis in complex biological samples.
研究领域:
生物传感器
·
分子诊断
·
核酸化学
·
肿瘤标志物
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NANOTECHNOLOGY
·
2021
·Vol. 33
·期 9
·095601
·
IF 3.874
Lulu Yu,Min Liu,Yanling Zhang,Yun Ni,Shaobo Wu,Ruijiang Liu
A magnetically induced self-assembly DNAzyme electrochemical biosensor based on gold-modified α-Fe2O3/Fe3O4 heterogeneous nanoparticles was successfully fabricated to detect Nickel(II) (Ni2+). The phase composition and magnetic properties of α-Fe2O3/Fe3O4 heterogeneous nanoparticles controllably prepared by the citric acid (CA) sol–gel method were investigated in detail. The α-Fe2O3/Fe3O4 heterogeneous nanoparticles were modified by using trisodium citrate as reducing agent, and the magnetically induced self-assembly α-Fe2O3/Fe3O4–Au nanocomposites were obtained. The designed Ni2+-dependent DNAzyme consisted of the catalytic chain modified with the thiol group (S1-SH) and the substrate chain modified with methylene blue (S2-MB). The MGCE/α-Fe2O3/Fe3O4–Au/S1/BSA/S2 electrochemical sensing platform was constructed and differential pulse voltammetry was applied for electrochemical detection. Under the optimum experimental parameters, the detection range of the biosensor was 100 pM–10 μM (R2 = 0.9978) with the limit of detection of 55 pM. The biosensor had high selectivity, acceptable stability, and reproducibility (RSD = 4.03%).
研究领域:
生物化学
·
纳米技术
·
环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Zehan Zeng,Mingzhen Zhong,Tong Liao,Xuan Zhou,Lin Shi,Xin Zhang,Ye Zhang,Jingyan Feng,Yiqian Chen,Shuya Sheng,Jun Chen,Lei Zheng,Weilun Pan,Jinxiang Chen
Plasma extracellular vesicles (EVs) have emerged as promising biomarkers for the early diagnosis of glioblastoma. However, rapid and efficient isolation and detection of tumor-specific EV subsets from plasma remains challenging. To address this, we developed an innovative integrated chip based on self-sacrificial metal–organic frameworks and the epidermal growth factor receptor (EGFR) aptamers called ZIF-8@EGFR aptamer (ZIF-8@Apt), which further coupled with a “nano-fuels”-driven DNAzyme amplification strategy. The core functionality of the constructed chip lies in its ability to capture EVs with the ZIF-8@Apt module. Once captured, numerous DNA “nano-fuel” structures are in situ assembled on the EVs’ surfaces. Subsequently, the ZIF-8@Apt@EV@nano-fuels composites are lysed to release Zn 2+ , the EVs’ nucleic acids, and additional nano-fuels for subsequent detection. Finally, a DNAzyme detection system is introduced, where the released nano-fuels activate the DNAzyme, leading to multiple signal amplification aided by Zn 2+ for highly sensitive detection of plasma EVs. This integrated approach effectively enriches and detects target EVs, achieving a significantly enhanced detection sensitivity of 500 particles/μL. In a clinical study involving glioblastoma patients, our chip demonstrated a remarkable ability to distinguish between cancer patients and healthy individuals, achieving an area under the curve (AUC) of 0.958. Moreover, the response signals of the chip can be conveniently read using fluorescence spectroscopy or microscope, offering a rapid, highly sensitive, and high-throughput analysis of plasma EVs for various application scenarios. Overall, this study introduces a comprehensive approach for the rapid isolation and accurate analysis of plasma EVs, holding significant promise for early diagnosis and disease monitoring.
研究领域:
生物医学工程
·
分子诊断
·
癌症研究
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）