MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Min Wang,Hui-Bin Wu,Xue Yang,Si-Xiang Zhang,Qin-Xin Wang,Guang-Xiang Luan,Hang Li,Zhi Zeng,Qiu-Long Zhang
A novel cascade dual-DNAzyme amplification platform was engineered. • The aptasensor utilized a non-competitive, strand displacement-free transduction mechanism. • The aptasensor exhibits excellent analytical performance with the LOD of 21.0 nM. • Practical applicability was validated in real food samples Tobramycin (TOB), a widely employed broad-spectrum antibiotic in animal husbandry, leaves residual traces that exert inevitable adverse effects on human health, including kidney impairment, neuromuscular blockage, respiratory paralysis, and cross-allergic reactions. Herein, we constructed a novel ligand-induced assembled E6-split DNAzyme cascaded 8–17 DNAzyme aptasensor and applied it to the highly sensitive detection of TOB. In this sensing design, we incorporated the aptamer sequence with a short stem into the input strand. In the absence of the target TOB, the structurally flexible aptamer-programmed input strand fails to form a stable complex with the E6-split DNAzyme. Upon binding of TOB to the aptamer, the input strand undergoes a transition to a compact and rigid conformation, thereby forming a stable three-strand complex consisting of the input strand, E6-split DNAzyme 1, and E6-split DNAzyme 2; this complex assembles a catalytically active domain and restores the substrate cleavage activity. Upon activation, the DNAzyme cleaves a rationally designed hairpin DNA containing an RNA cleavage site to release an extra 8–17 DNAzyme sequence. The liberated DNAzyme subsequently drives the continuous cleavage of fluorophore-quencher labeled reporter probes, resulting in the progressive accumulation of fluorescence signals. Under the optimized reaction conditions, the detection range displays a well-defined linear relationship from 100 nM to 200 μM, with a limit of detection (LOD) of 21.0 nM. By circumventing equilibrium constraints and obviating protein enzymes or nanomaterial modifications, this cascade DNAzyme architecture offers a versatile, sequence-independent, and robust strategy for small-molecule antibiotic monitoring, with broad potential for adaptation to diverse non-nucleic acid targets in food safety. Download: Download high-res image (135KB) Download: Download full-size image
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2026
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IF 8.7
Wei Guo,Mingzhu Fan,Zhisheng Sun,Huakui Huang,Guichun Mo,Zhihui Luo,Jiayao Xu,Ling Liang,Shulong Wang
Abnormal protein expression is closely linked to the onset and progression of various diseases, making real-time in vivo monitoring of proteins highly significant for disease diagnosis. Herein, a biodegradable and cofactor self-sufficient multifunctional DNAzyme bioinspired mineralization nanodevice has been developed for catalytic amplified imaging of low-abundance protein in vivo. The nanodevice was fabricated by in situ encapsulation of two antibody-labeled nucleic acid strands containing the target protein epitope sequence and partial split DNAzyme sequences (P1 and P2), together with a molecular beacon (H1) labeled with a fluorophore and quencher, into pH-responsive manganese-doped calcium carbonate nanoparticles. After accumulating at tumor sites and being internalized by cells, the nanodevice disintegrates in the weakly acidic intracellular environment. The target protein specifically recognizes the antibody-conjugated P1 and P2, thereby generating a complete DNAzyme sequence. Moreover, Mn 2+ ions released from the nanodevice activate the DNAzyme, which cleaves molecular beacon to restore fluorescence. The cleaved molecular beacon products can rehybridize with P1 and P2 to trigger iterative reactions, thereby converting protein amplification analysis into nucleic acid amplification analysis and enabling sensitive in vivo imaging of low-abundance proteins. Using vascular endothelial growth factor (VEGF) as a model, the detection limit of the nanodevice for VEGF reached 24.7 fM. Furthermore, in vivo fluorescence imaging of VEGF enabled clear discrimination between tumor and normal tissues. By simply replacing the corresponding aptamer sequences and antibody molecules, the developed nanodevice can be readily extended to detect other biomolecules, providing a promising tool for highly sensitive in vivo imaging of low-abundance biomolecules.
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高影响因子
ACS Applied Materials & Interfaces
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2024
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IF 8.3
Yinbo Ban,Fu Zhou,Hao Wang,Fuqiang Zhang,Mengmeng Xia,Yifei Wan,Suwan Yang,Rong Liu,Xiayan Wang,Guangfeng Wang
Endowing current artificial chemical reactions (ACRs) with high specificity and intricate activation capabilities is crucial for expanding their applications in accurate bioimaging within living cells. However, most of the reported ACR-based evaluations relied on either single biomarker stimuli or dual activators without obvious biological relevance, still limiting their accuracy and fidelity. Herein, taking the metal-ion-dependent DNAzyme cleavage reaction as a model ACR, two regulators, glutathione (GSH) and telomerase (TE) activated DNAzyme cleavage reactions, were exploited for precise discrimination of cancerous cells from normal cells. DNA probe was self-assembled into the ZIF-90 nanoparticle framework to construct coordination-driven nanoprobes. This approach enhances the stability and specificity of tumor imaging by utilizing biomarkers associated with rapid tumor proliferation and those commonly overexpressed in tumors. In conclusion, the research not only paves the way for new perspectives in cell biology and pathology studies but also lays a solid foundation for the advancement of biomedical imaging and disease diagnostic technologies.
研究领域:
生物医学工程
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分子生物学
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癌症研究
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纳米技术
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细胞生物学
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疾病诊断
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PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Manlin Luo,Chengyan Yin,Wei Liu,Kerage Dorothy Mokeira,Meng Luo,Ling Tang,Jing Liu,Shuxia Xu
In this study, a Ce13d DNAzyme-based electrochemical biosensor for the detection of sodium ions (Na + ) in the serum was developed. The indium tin oxide (ITO) electrode was modified with Au gold nanoparticles (AuNPs), then immobilized with double-stranded DNA formed by thiol of substrate strand and enzyme strand bound via Au-S link. In the presence of Na + , the resulted Ce13d DNAzyme could preferentially bind Na + , and induced local folding of Ce13d to form a G-quadruplex-like structure, which enhanced binding ability to methylene blue (MB). By using the specificity of DNAzyme and change of its conformation, the quantitative detection of Na + could be achieved. Thus, the developed sensing platform could identify Na + with a linear response range of 10 nM-100 mM and a detection limit of 1.03 nM. The electrochemical biosensor also exhibited excellent selectivity for Na + in the presence of five interfering ions, including K + , Cu 2+ , Mg 2+ , Zn 2+ and Ca 2+ . Furthermore, the proposed biosensor showed satisfactory results for Na + in serum sample analysis, which highly correlated with the inductively coupled plasma atomic emission spectrometry method. This work demonstrated a simple and reliable method for Na + in serum, which opens the way for potential applications in disease diagnosis and environmental testing.
研究领域:
生物化学
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医学检测
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分析化学
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Huinan Chen,Jingyao Song,Yuanyuan Li,Dongmei Deng,Yuchen Song,Xiaoli Zhu,Liqiang Luo
Tumor biomarkers are a kind of important biomolecules for aiding cancer diagnosis and assessing tumor efficacy, among which alpha-fetoprotein (AFP) is a crucial tumor biomarker for the early diagnosis of hepatocellular carcinoma. In this study, a novel cascade signal amplifying strategy was designed for the ultrasensitive detection of AFP by DNAzyme cleaving-mediated hybridization chain reaction (HCR). The primary antibody was immobilized on the Au nanoparticles modified glassy carbon electrode, which subsequently captured AFP and the secondary antibody. Then, streptavidin was combined with the biotin-modified secondary antibody and Cu 2+ -specific DNAzyme. When Cu 2+ was present, the substrate strand of the DNAzyme would cleave and produce an ssDNA end. Finally, HCR was triggered to form long double-stranded DNA on the electrode, which would greatly inhibit the charge transfer at the electrode/solution interface. Therefore, the DNAzyme cleaving-mediated HCR strategy can be applied as cascade signal amplification for the ultrasensitive electrochemical determination of tumor biomarker AFP. Under optimal conditions, the immunosensor exhibits a broad linear range of 0.001−100 ng·mL −1 (R 2 = 0.990) and a low detection limit of 15.8 fg·mL −1 . The proposed sensor displays high sensitivity and selectivity for AFP detection, which can be employed as a universal sensing platform in clinical diagnostics.
研究领域:
生物医学工程
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分析化学
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癌症诊断
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高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2026
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IF 8.7
Siyu Wang,Jiaqiong Wu,Dongdong Zhang,Xiao Li,Jianqiong Pan,Liuqin Teng,Qingao Meng,Chengkai Zhuang,Jiahui Xu,Ting Chen,Shanshan Chen,Xiahui Lin,Shanni Hong
To disrupt the glycolytic metabolism vital for pancreatic cancer (PC), we developed a cascade-amplified and PC-targeted nanosystem, HMAD@EP-XQ2d. It integrates a DNAzyme for GLUT1 gene silencing, AuNPs-based nanozymes for catalytic activity, and a HMnO 2 carrier, cloaked in an aptamer-modified erythrocyte membrane. The system actively targets CD71-overexpressing PC cells. Upon accumulation in the tumor microenvironment (TME), the HMnO 2 degrades, initiating a therapeutic cascade: the released Mn 2+ serves a dual function-they activate the DNAzyme for upstream blockade of glucose uptake, and act as a contrast agent for magnetic resonance imaging (MRI), enabling real-time monitoring of drug delivery and accumulation; concurrently, the exposed gold nanozymes exert glucose oxidase-like (GOx-like) activity to deplete glucose (downstream depletion) and peroxidase-like (POD-like) activity to convert the resultant H 2 O 2 into cytotoxic hydroxyl radicals (oxidative stress). This three-pronged, synergistic attack on glycolysis induces a severe energy crisis in PC cells. Both in vitro and in vivo studies demonstrate potent antitumor efficacy, favorable biocompatibility, and MRI contrast capability. By integrating gene silencing, enzymatic catalysis, and imaging into a single responsive system triggered by the TME, this all-in-one theranostic platform offers a novel precise metabolic intervention strategy for pancreatic cancer.
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PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
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2026
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IF 5.3
Sha Chen,XinJun Zhang,Xia Chen,Ying Tang,Yao Chen,YingQi Wang,Tong Wang,DeHuan Yang
Aflatoxin B1 (AFB1) is a hazardous mycotoxin commonly found in agricultural products, necessitating sensitive and reliable detection methods. In this work, a novel aptamer-based fluorescence sensing strategy was developed, in which DNAzyme acts as both signal generator and amplifier. In the absence of AFB1, a blocking strand hybridizes with the DNAzyme arm, inhibiting its catalytic activity. When AFB1 is present, the aptamer specifically binds the target, releasing the blocking strand and restoring DNAzyme activity. The activated DNAzyme cleaves a fluorophore–quencher labeled substrate, leading to enhanced fluorescence. To achieve accurate quantification, a probe technique-based generalized multivariate standard addition strategy (GMSAprobe) was employed. The method enabled reliable determination of AFB1 in peanut samples with recoveries of 96–102% and a detection limit of 0.56 μg/kg, consistent with results from a commercial ELISA kit and well below regulatory limits.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2020
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IF 7.1
Long Wang,Huali Jin,Min Wei,Wenjie Ren,Yurong Zhang,Liying Jiang,Tao Wei,Baoshan He
Herein, an electrochemical aptasensor was constructed for ultra-sensitive determination of T-2 toxin based on Silver ion (Ag + ) -dependent DNAzyme-assisted signal amplification and Gold nanoparticles/Graphene oxide @ manganese dioxide (AuNPs/MnO 2 @GO) nanocomposites. In the presence of Ag + , the secondary structure of the T-2 aptamer is altered via the Ag + mediated coordination of C–Ag + –C base pairs. When T-2 toxin is added, the secondary structure of DNA is changed again, and the trapped Ag + are released. Then, the DNAzyme could repeatedly bind and cleave methylene blue (MB)-modified substrate DNA (S-DNA) in the assistance of Ag + , which leads to a decrease in the electrochemical signal. With the optimal conditions, there is a good linear relationship between the current value and the logarithm of T-2 toxin concentration in the range from 2 fg mL −1 to 20 ng mL −1 , and a detection limit of 0.107 fg mL −1 is obtained. Moreover, the proposed method was successfully applied to measure T-2 toxin in beer, thus showing great potential of this system for food safety and quality control.
研究领域:
分析化学
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生物传感器
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毒理学
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食品科学
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高影响因子
TALANTA
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2021
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IF 6.057
Ting Huang,Guixun Chen,Birong Liu,Zizhong Yang,Yuanwei Huang,Baoping Xie,Min-Min Li,Jin-Xiang Chen,Jun Chen,Zong Dai
Sensitive, specific and rapid methods for detecting microRNAs (miRNAs) play critical roles in disease diagnosis and therapy. Enzyme-free amplification techniques based on DNAzyme assembly have recently been developed for the highly specific miRNA analysis. However, traditional DNAzyme-based assembly (free DNAzyme) amplifiers is mainly dependent on the target-induced split DNAzyme fragments to assemble into activated DNAzyme structures, which have made a compromise between the sensitivity and specificity due to the random diffusion of dissociative probes in a bulk solution with poor kinetics. Herein, based on a rationally designed DNA probe , we developed an intramolecular DNAzyme assembly (intra-DNAzyme) method to overcome these challenges. The miR-373 is used as model analyte for our current proof-of-concept experiments. Compared with the free-DNAzyme method, our method showed significantly improved analytical performance in terms of dynamic range, assay sensitivity and speed. This method can detect miR-373 specifically with a detection limit as low as 4.3 fM, which is about 83.7 times lower than the previous free-DNAzyme method. This intra-DNAzyme strategy would be of great value in both basic research and clinical diagnosis.
研究领域:
分子生物学
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生物技术
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纳米技术
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诊断学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2018
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IF 5.123
Binxiao Li,Jing Liu,Hong Zhou
An amplified fluorescence biosensing strategy for serum prostate specific antigen (PSA) was developed on the basis of DNAzyme. In presence of cofactor Zn 2+ , Zn 2+ -dependent DNAzyme could cleave the hairpin substrate probes which were dispersed in solution and generate remarkable fluorescent signal. Taking advantage of the magnetic beads as a carrier, one target protein could bring plentiful hairpin substrate probes on to the electrode through a sandwich structure (Ab 1 /PSA/biotin-Ab 2 ). Moreover, during the cleavage process of as formed DNAzyme, DNAzyme did not be destroyed and could further react with other hairpin probes, then generated continuous fluorescent signal. Benefited by this amplified strategy, the limit of detection (LOD) was low to 0.05 ng mL −1 , which was much lower than our previous reports. This method could be applied to detect different protein biomarkers in serum without corresponding aptamers by changing the corresponding antibodies and thus showed a remarkable prospect in clinical application.
研究领域:
生物传感
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分析化学
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分子生物学
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临床诊断
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DOI
PubMed
高影响因子
TALANTA
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2018
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IF 4.244
Ke Li Lin,Tong Yang,Hong Yan Zou,Yuan Fang Li,Cheng Zhi Huang
Here we first reported that graphitic carbon nitride nanosheet (g-C 3 N 4 NS) could effectively quench the chemiluminescence (CL) of luminol-hydrogen peroxide (H 2 O 2 ) system. According to the new discovery, a label-free and homogeneous CL aptasensing platform was designed for sensitive detecting of biomarkers. In the absence of target, DNA probe containing hemin/G-quadruplex DNAzyme structure was adsorbed on the surface of g-C 3 N 4 NS, causing the CL quenching of luminol through an electron transfer process. However, in the presence of the target, a DNA-DNA duplex was formed due to DNA hybridization reaction and target recognition effect, which could not be adsorbed onto the g-C 3 N 4 NS surface because of its weak affinity. Thus, the electron transfer was blocked and the CL emission of luminol could be enhanced. The proposed CL aptasensor could detect carcinoembryonic antigen (CEA) with a detection limit of 63.0 pg/mL and it can also be used as a general detecting strategy for adenosinetriphosphate (ATP) detection. This aptasensing platform exhibited high sensitivity toward biomarkers and the probe need not be labeled, showing great promise for disease diagnosis.
研究领域:
生物化学
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2017
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IF 5.401
Yongmei Wu,Gaiping Li,Lina Zou,Sheng Lei,Qian Yu,Baoxian Ye
Herein, we developed a high-performance electrochemical aptasensor for the detection of carcinoembryonic antigen (CEA) using highly active peptide-conjugated hemin/G-quadruplex (hGQ-peptide) and porous Pd nanoparticles (PdNPs). Of note, the hGQ-peptide showed an enhanced catalytic activity as compared to the unmodified hGQ, and thus served as efficient electrocatalyst in this biosensor. Porous PdNPs acted as supported matrixes for high immobilization of hGQ-peptide hybrids, aptamer, redox-active toluidine blue (Tb) and alcohol dehydrogenase (ADH), resulting in the formation of ADH/Tb/hGQ-peptide/aptamer/PdNPs as the proposed bioprobes. Upon the sandwich-type specific reaction between aptamer and CEA, the bioprobe was presented on the sensing interface. Highly effective signal amplification could be then achieved by the bioprobe catalyzed cascade enzymatic reaction. Compared with the original hGQ, the hGQ-peptide yielded more than 2-fold improvement in signal amplification because of the peptide-enhanced catalysis. Therefore, the designed aptasensor displayed a wide linear range from 0.0001 to 100 ng mL −1 with a lower detection limit of 20 fg mL −1 for CEA determination, presenting the better analytical performance as compared to the relevant prior biosensors. This work further expanded the application of hGQ DNAzyme in biosensor. With the good anti-interference ability and selectivity, the proposed electrochemical aptasensor held great promise for ultrasensitive detection of other proteins in clinical diagnosis.
研究领域:
生物化学
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分析化学
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生物医学工程
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分子生物学
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高影响因子
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