RSC Advances
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2018
·Vol. 8
·期 45
·25611-25616
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IF 2.936
Dan Han,Chunying Wei
In this paper, we construct a fluorescence nucleic acid probe based on DNA-templated silver nanoclusters (DNA-Ag NCs) for the detection of the p53 gene. The fluorescence biosensing of the “turn-on” model is successfully implemented as a result of the target-triggered configurational change in the hairpin DNA probe and the synthesis of fluorescent Ag NCs. With this biosensor, the limit of detection (LOD) for the p53 gene is 3.57 nM and the linear range is 250–2500 nM in phosphate buffer solution, while a LOD of 6.06 nM in the linear range of 250–2500 nM is obtained in 1% diluted fetal calf serum. So this probe, with its advantages of specificity, practical application, easy operation and low cost, will have favourable development prospects in biological sensing and imaging.
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Journal of Materials Chemistry B
·
2018
·Vol. 6
·期 36
·5718-5728
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IF 4.776
Jiayi Song,Hao Shen,Ye Yang,Zixin Zhou,Ping Su,Yi Yang
The overall stability and activity of immobilized enzyme systems have suffered from non-specific adsorption immobilization of enzymes at the surface of carriers through electrostatic and hydrophobic interactions. Elimination of these non-specific adsorptions of enzymes on the surface of carriers is critical for enzyme reactions. Herein, for the first time, we have prepared zwitterion-functionalized magnetic particles with amino, phosphonate, and thiol functional groups to coimmobilize glucose oxidase and horseradish peroxidase by DNA directed immobilization. The zwitterionic surface of the multifunctional magnetic particles could efficiently suppress non-specific adsorption of different kinds of enzymes, and the immobilized multienzyme catalyst without non-specifically adsorbed enzymes exhibited excellent enzymatic activity, stability, and reusability compared with those of free and non-specifically adsorbed enzymes. The immobilized multienzymes maintained 87% of their enzymatic activity after two weeks of storage at 4 °C and 58% of their enzymatic activity after 90 min of thermal incubation at 60 °C. Furthermore, the immobilized multienzymes exhibited more than 87% enzymatic activity after reuse for 10 cycles. The apparent Km and catalytic efficiency (kcat/Km) values of the immobilized multienzymes were 12.6 mM and 4.03 s−1 mM−1, respectively, which were 0.2- and 11.8-fold better than those of the free enzymes, indicating effective cascade efficiency and substrate affinity. The optimal ratio of GOx and HRP was 1 : 5, and the prepared immobilized enzymes could detect low concentrations of glucose (0.5 μM) with excellent selectivity. Therefore, we believe that the strategy developed in this study can be widely applied in biotechnology, industrial catalysis, and biomedical engineering.
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Journal of Nanomaterials
·
2017
·
IF 1.871
Juan Zhang,Huanhuan Weng,Xiangwan Miao,Quanming Li,Siqi Wang,Huifen Xie,Tao Liu,Minqiang Xie
A novel folate (FA) receptor-targeted superparamagnetic Fe3O4 nanoparticles (SPIONs) codelivering cisplatin (CDDP) and tissue factor pathway inhibitor-2 (TFPI-2) plasmid DNA (pDNA) was constructed. The core shell nanocomposites ([email protected]) were composed of superparamagnetic Fe3O4 core that binds CDDP and TFPI-2 shell that combines with folate-polyethylene glycol-polyethyleneimine (FA-PEG-PEI) via electrostatic interaction. The shell containing FA-PEG-PEI and TFPI-2 plasmid was synthesized through amidation reaction and electrostatic adsorption and the core containing SPION-CDDP was modified by aldehyde sodium alginate. Proton nuclear magnetic resonance and Fourier transform infrared spectra on FA-PEG-PEI polymers showed characteristic peaks of various metabolites in corresponding frequency. Transmission electron microscopy image of [email protected] nanoparticles demonstrated a near-monodisperse spherical morphology, while dynamic light scattering studies indicated an intensity-average diameter of 149.5 nm. Zeta potential was 14.89 ± 1.83 mv and the final concentration of loaded CDDP was 100 ug/ml. Gel electrophoresis data showed that the nanocomposite would protect TFPI-2 pDNA from being digested by DNases. Compared with CNE-2 cells, the good targetability and better gene transfection efficiency (57.9%) were detected by Prussian blue iron stain and fluorescence analysis in HNE-1 cells in vitro. The results suggested the potential application of [email protected] as a multifunctional anticancer nanomedicine on targeting therapy for FR positive NPC.
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2017
·
IF 2.536
Jie-Hua Shi,Kai-Li Zhou,Yan-Yue Lou,Dong-Qi Pan
Molecular interaction of darunavir (DRV), a HIV protease inhibitor with calf thymus deoxyribonucleic acid (ct-DNA) was studied in physiological buffer (pH 7.4) by multi-spectroscopic approaches hand in hand with viscosity measurements and molecular docking technique. The UV absorption and fluorescence results together revealed the formation of a DRV–ct-DNA complex having binding affinities of the order of 10 3 M − 1 , which was more in keeping with the groove binding. The results that DRV bound to ct-DNA via groove binding mode was further evidenced by KI quenching studies, viscosity measurements, competitive binding investigations with EB and Rhodamine B and CD spectral analysis. The effect of ionic strength indicated the negligible involvement of electrostatic interaction between DRV and ct-DNA. The thermodynamic parameters regarding the binding interaction of DRV with ct-DNA in terms of enthalpy change (Δ H 0 ) and entropy change (Δ S 0 ) were − 63.19 kJ mol − 1 and − 141.92 J mol − 1 K − 1 , indicating that hydrogen bonds and van der Waals forces played a predominant role in the binding process. Furthermore, molecular simulation studies suggested that DRV molecule was prone to bind in the A-T rich region of the minor groove of DNA.
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Journal of Materials Chemistry B
·
2024
·
IF 6.1
Mingsan Miao,Liang Guo,Jinyan Xue,Yuzhen Jia,Zhenzhen Cui,Huai-Xia Yang
The development of a simple, rapid, and sensitive technology for the simultaneous detection of mycotoxins is of great significance in ensuring the safety of foods and drugs. Herein, a fluorescence aptasensor with high sensitivity and reproducibility for the simultaneous detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA) was developed. In this sensing system, AFB1 and OTA aptamers were co-immobilized on the surface of magnetic beads (MBs) to form a Y-shaped structure through the principle of complementary base pairing, which were used as recognition probes to specifically capture the target. Activators generated by electron transfer for atom transfer radical polymerization (ARGET ATRP) was used as signal amplification strategy to improve the sensitivity. The initiator modified at the end of the antibody initiates the ARGET ATRP reaction. Different fluorescence signals were designed to achieve simultaneous detection of OTA and AFB1 with the limit of 18.62 and 1.60 fg/mL for AFB1 and OTA, respectively. In addition, experiments were conducted on three types of samples, and the recoveries of the two mycotoxins ranged from 87.30% to 109.50%, with relative standard deviations ranging from 0.50% to 4.92% under reproducible conditions. The results suggest that the developed aptasensor is sufficient to meet the different regulatory requirements of the two mycotoxins in food and drug safety and shows great potential.
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FOOD CHEMISTRY
·
2024
·
IF 8.5
Kai Wang,Xiaohai Yan,Jiamin Wu,Jianrui Qi,Mengge Ning,Menghui Li,Rui Sun,Zhouli Wang,Yahong Yuan,Tianli Yue
Deoxynivalenol (DON) contamination in cereals is a widespread issue with global implications, necessitating the development of efficient detection methods. Here, a fluorescent aptasensor integrating target-responsive DNA three-way junction (TWJ) and DNA walking machine was developed to detect DON. The DON-specific aptamer (Apt) and the walker (Walker DNA) are integrated into TWJs. TWJs initiate the DNA walking machine with the assistance of Nb. Bbv CI to provide signal amplification for sensing events. The constructed fluorescent aptasensor selectively recognizes and detects DON. Additionally, the sensing platform explored the quantitative relationship between fluorescence signal changes and DON concentration, achieving a limit of detection (LOD) of 0.009 ng/mL. Ultimately, the method performed well in the detection of cornmeal samples with recoveries ranging from 95.49 % to 100.82 %. The method demonstrates good stability and is expected to be a potential candidate for the accurate detection of DON in grains.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Luting Yan,Yuhao Zhang,Qi Guo,Lei Tian,Falan Li,Qingqing Yang,Yanyan Zhang
Most electrochemiluminescent (ECL) biosensors currently necessitate coreactants that affect sensor detection accuracy and increase sensor construction complexity. This study detailed the creation of an ECL biosensor without coreactant, utilizing Methionine-capped Au nanoclusters (Met-AuNCs) as luminophore and a Pb 2+ -dependent DNAzyme-mediated DNA walker signal amplification strategy to detect Vibrio parahaemolyticus (Vp) in scallops. Double-stranded DNA (dsDNA) was produced through hybridization of the Vp aptamer with DNA walker. The electrode surface was loaded with cDNA labeled with Met-AuNCs. Upon the presence of Vp, it competitively bound to the aptamer, resulting in dsDNA dissociation and exposing the DNAzyme sequence in the DNA walker. The introduction of Pb 2+ initiated the activation and circulation of the DNAzyme-mediated DNA walker on the electrode surface, causing the cDNA to be sheared. As a result, the cDNA-Met-AuNCs on the surface of the electrode decreased, resulting in a reduction of ECL signal intensity. The ECL biosensor detected Vp at a range of 1.00×10 1 -1.00×10 7 CFU/mL, under optimal conditions, with the limit of detection (LOD) being 1.23 CFU/mL. In this work, the biosensor demonstrated excellent stability and specificity and had proven successful in detecting actual samples of aquatic products. This study offers a novel ECL method without a coreactant for the detection of foodborne pathogens in aquatic products.
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BIOSENSORS & BIOELECTRONICS
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2024
·
IF 10.7
Mei Zhao,Jingyi Guo,Zilin Chen,Fang Wang
5-formylcytosine (5 fC) and 5-carboxylcytosine (5caC) serve as key intermediates in DNA demethylation process with significant implications for gene regulation and disease progression . In this study, we introduce a novel electrochemical sensing platform specifically designed for the sensitive and selective detection of 5 fC and 5caC in DNA . Protein A-modified magnetic beads (ProtA-MBs) coupled with specific antibodies facilitate the immunorecognition and enrichment of these modified bases. Signal amplification is achieved through several chemical reactions involving the interaction between N 3 -kethonaxl and guanine , copper-free click chemistry for the attachment of dibenzocyclooctyne (DBCO)-Biotin, and the subsequent recognition by streptavidin-conjugated horseradish peroxidase (SA-HRP). The assay's readout is performed on a disposable laser-induced graphene (LIG) electrode, modified with the bead-antibody-DNA complex in a magnetic field , and analyzed using differential pulse voltammetry in a system employing hydroquinone (HQ) as the redox mediator and H 2 O 2 as the substrate. This immunosensor displayed excellent sensitivity, with detection limits of 14.8 fM for 5 fC across a 0.1–1000 pM linear range and 87.4 fM for 5caC across a 0.5–5000 pM linear range, and maintained high selectivity even in the presence of interferences from other DNA modifications . Successful application in quantifying 5 fC and 5caC in genomic DNA from cell extracts, with recovery rates between 97.7% to 102.9%, underscores its potential for clinical diagnostics. N 3 -kethoxal was used for the first time in an electrochemical sensor. This work not only broadens the toolkit for detecting DNA modifications but also provides a fresh impetus for the development of point-of-care testing (POCT) technologies.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Yi Wang,Zelong Chen,Xingwang An,Jiahui Li,Jiaxuan Li,Zhichao Pei,Yuxin Pei
DNA damage-based therapy is widely used in cancer treatment, yet its therapeutic efficacy is constrained by the redox homeostasis and DNA damage repair mechanisms of tumor cells. To address these limitations and enhance the efficacy of DNA damage-based therapy, HA-CuH@MTX, a copper-histidine metal-organic complex (CuH) loaded with methotrexate (MTX) and modified with hyaluronic acid (HA), was developed to amplify the DNA damage induced. In vitro experiments demonstrated that the presence of both Cu + and Cu 2+ in HA-CuH@MTX enables two-way regulated redox dyshomeostasis (RDH), achieved through Cu + -catalyzed generation of •OH and Cu 2+ -mediated consumption of glutathione, thereby facilitating efficient DNA oxidative damage. In addition, DNA damage repair is synergistically inhibited by impairing nucleotide synthesis via histidine metabolism and MTX downregulation of tetrahydrofolate, a crucial raw material in nucleotide synthesis. In vivo experiments with 4T1 tumor-bearing mice demonstrate 83.6 % inhibition of tumor growth by HA-CuH@MTX. This work provides a new strategy to amplify the DNA damage caused by DNA damage-based cancer therapies, and holds great potential for improving their therapeutic efficacy.
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STARCH-STARKE
·
2015
·
IF 1.677
Xuan Liang,Shuyun Wu,Xianyue Ren,Daping Quan,Li-Ming Zhang,Yubing Deng,Liqun Yang
A series of hyperbranched glycogen derivatives conjugated with different contents of cationic 3-(dimethylamino)-1-propylamine residues, named DMAPA-Glyp derivatives, were synthesized at room temperature. Resonance light scattering (RLS) spectroscopy was used to investigate the complexation behavior and efficacy of the DMAPA-Glyp derivatives and plasmid DNA (pDNA). Results indicated that the DMAPA-Glyp derivatives and pDNA molecules began to aggregate and form complexes at a weight ratio of 2. Following further increases in the DMAPA-Glyp derivatives (w/w >2), pDNA was condensed to form compact complexes. RLS spectroscopy is thus a useful tool for studying the complexation behavior and efficacy of cationic polymers with nucleic acid molecules. Circular dichroism analysis further revealed that the conformation of pDNA transformed from B-form to C-form and A-form during the complexation process.
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FOOD CONTROL
·
2025
·
IF 6.3
Xinru Ren,Weiqing Sun,Bowen Li,Yijing Xiao,Xudong Yue,Meili Yang,Qing-ao Pang,Rui Zhu,Zhiqiang Guo,Haiping Zhang,Yu Wang,Su Liu,Jiadong Huang
The detection of pathogenic bacteria plays a crucial role in ensuring food safety and public health. The integration of isothermal amplification technology with the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) system offers a versatile approach to detecting foodborne pathogens. However, this method is often constrained by the incompatibility of the reaction systems and the lack of control over CRISPR/Cas activity. Here, we demonstrated a fluorescence biosensing strategy based on linear causal-regulated and autocatalysis-driven DNA circuits (termed LC@ADC) for highly sensitive and specific detection of Salmonella Typhimurium ( S. Typhimurium ). The approach of combining hybridization chain reaction (HCR) with DNAzyme-driven CRISPR/Cas12a activation was proposed for the first time, enabling not only one-step cascade signal amplification but also controllable DNA circuits. The LC@ADC strategy is capable of quantifying concentrations of S. Typhimurium ranging from 10 to 1 × 10 5 CFU/mL, with a detection limit as low as 6.53 CFU/mL. Furthermore, the LC@ADC strategy has been successfully employed for the detection of S. Typhimurium in samples of orange juice, chicken, and lake water. Therefore, this strategy provides a novel approach for the high-performance detection of pathogenic bacteria.
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Frontiers in Plant Science
·
2017
·
IF 4.291
Zha Liangping,Liu Shuang,Liu Juan,Jiang Chao,Yu Shulin,Yuan Yuan,Yang Jian,Wang Yaolong,Huang Luqi
The content of active compounds differ in buds and flowers of Lonicera japonica (FLJ) and L. japonica var. chinensis (rFLJ). Chlorogenic acid (CGAs) were major active compounds of L. japonica and regarded as measurements for quality evaluation. However, little is known concerning the formation of active compounds at the molecular level. We quantified the major CGAs in FLJ and rFLJ, and found the concentrations of CGAs were higher in the buds of rFLJ than those of FLJ. Further analysis of CpG methylation of CGAs biosynthesis genes showed differences between FLJ and rFLJ in the 5′-UTR of phenylalanine ammonia-lyase 2 (PAL2). We identified 11 LjbZIP proteins and 24 rLjbZIP proteins with conserved basic leucine zipper domains, subcellular localization, and electrophoretic mobility shift assay showed that the transcription factor LjbZIP8 is a nuclear-localized protein that specifically binds to the G-box element of the LjPAL2 5′-UTR. Additionally, a transactivation assay and LjbZIP8 overexpression in transgenic tobacco indicated that LjbZIP8 could function as a repressor of transcription. Finally, treatment with 5-azacytidine decreased the transcription level of LjPAL2 and CGAs content in FLJ leaves. These results raise the possibility that DNA methylation might influence the recruitment of LjbZIP8, regulating PAL2 expression level and CGAs content in L. japonica.
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高影响因子
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