Biosensors-Basel
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2022
·Vol. 12
·期 9
·745
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IF 5.743
Li Gao,Yixi Deng,Haolu Liu,King Solomon,Bianjiang Zhang,Huimei Cai
Tea is an important economic crop and health beverage in China. The presence of heavy metal ions in tea poses a significant threat to public health. Here, we prepared an aptamer biosensor labelled with AIEgen nanospheres to detect Pb2+in tea. The dsDNA modified by amino and phosphoric acid was combined with the carboxylated AIEgen NPs to form AIEgen-DNA with a fluorescence group, which was then fixed to the surface of Zr-MOFs to quench the fluorescence of AIEgen NPs. At the same time, PEG was added to remove nonspecific adsorption. Then Pb2+was added to cut the DNA sequences containing the cutting sites, and AIEgen NPs and part of the DNA sequences were separated from the Zr-MOFs surface to recover the fluorescence. By comparing the fluorescence changes before and after adding Pb2+, the detection limit of Pb2+can reach 1.70 nM. The fluorescence sensor was applied to detect Pb2+in tea, and the detection results showed that the tea purchased on the market did not contain the concentration of Pb2+within the detection range. This study provides new insights into monitoring food and agriculture-related pollutants based on fluorescent biosensors.Keywords:lead ions;fluorescent biosensors;metal-organic frames;tea
研究领域:
分析化学
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食品科学
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环境科学
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产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
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2023
·
IF 9.8
Junhua Chen,Manjia Chen,Hui Tong,Fei Wu,Yizhang Liu,Chengshuai Liu
A fluorescence biosensor was developed for the ultrasensitive detection of the available lead in soil samples by coupling with DNAzyme and hairpin DNA cyclic assembly. The biorecognition between lead and 8-17 DNAzyme will cleave the substrate strands (DNA2) and release the trigger DNA (T), which can be used to initiate the DNA assembly reactions among the hairpins (H1, H2, and H3). The formed Y-shaped sensing scaffold (H1-H2-H3) contains active Mg 2+ -DNAyzmes at three directions. In the presence of Mg 2+ , the BHQ and FAM modified H4 will be cleaved by the Mg 2+ -DNAyzme to generate a high fluorescence signal for lead monitoring. The linear range of the fluorescence biosensor is from 1 pM to 100 nM and the detection limit is 0.2 pM. The biosensor also exhibited high selectivity and the nontarget competing heavy metals did not interfere with the detection results. Compare with the traditional method (DTPA+ICP-MS) for the available lead detection, the relative error (Re) is in the range from −8.3 % to 9.5 %. The results indicated that our constructed fluorescence biosensor is robust, accurate, and reliable, and can be applied directly to the detection of the available lead in soil samples without complex extraction steps.
研究领域:
环境科学
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分子生物学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Bowen Xiong,Xiang Li,Yuting Zhang,Changling Yan,Gongke Wang
The overexpression of vascular endothelial growth factor 165 (VEGF 165 ) in cancer cells plays a pivotal role in promoting tumor metastasis by facilitating their excessively rapid proliferation and division. Hence, the development of analytical methods possessing high sensitivity and resistance to interference is imperative for the detection of VEGF 165 . Various types of aptasensors have been devised for VEGF 165 detection; however, the performance of these biosensors can be influenced by non-target signals caused by conformational changes in unbound aptamers. The paper shows the creation of a precise and sensitive fluorescence biosensor designed to detect VEGF 165 by using a VEGF 165 -specific split aptamer. Additionally, this biosensor employs nicking enzyme-assisted DNA walker coupling with CRISPR-Cas12a to achieve dual-signal amplification. The VEGF 165 calibration curve shows a detection limit of 268 fM and has a broad linear range from 5 to 4000 nM. The fluorometric biosensor was utilized to detect VEGF 165 in human serum and cellular homogenate samples, yielding good outcomes. The innovative design serves as proof of concept and demonstrates significant potential in detecting various targets.
研究领域:
生物技术
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2016
·
IF 4.758
Ling Zhu,Guanhong Xu,Quan Song,Tang Tang,Xu Wang,Fangdi Wei,Qin Hu
In this paper, a novel fluorescent biosensor based on DA aptamer labeled carbon dots (aptamer-CDs) and nano-graphite (NG) for the determination of dopamine (DA) was developed. In this system, aptamer-CDs were in charge of energy donor (fluorophor) and chemical recognition, while NG served as an energy acceptor. In the absence of DA, aptamer-CDs were adsorbed on the surface of NG through π-π stacking and hydrophobic interaction, which led to the fluorescence quenching of aptamer-CDs due to the surface energy transfer. When it was in presence, DA combined with its aptamer anchored on the surface of CDs to form specific globular chain structure which would detach from the surface of NG and thus the fluorescence intensity of aptamer-CDs was recovered. Under the optimal conditions, the fluorescence intensity of aptamer-CDs increased linearly with the increase of DA concentration in the range of 0.10–5.00 nM and the limit of detection was 0.055 nM. This method was successfully applied to the determination of DA in human urine samples with satisfactory results.
研究领域:
分析化学
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生物传感技术
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医学检测
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Hui Yue,Jinri Chen,Xu Chen,Xiaoli Wang,Yuting Zhang,Nandi Zhou
Sialic acids (SAs), particularly its most common derivative N-acetylneuraminic acid (Neu5Ac) are involved in many biological processes in vivo . Thus it is of great significance to detect Neu5Ac and trace the sialylation of glycoproteins and other bio-active molecules. Aptamers are molecular elements for bio-recognition with high affinity, which have been widely used in bioanalysis and imaging. However, the ideal aptamers for saccharides have seldom been reported. Magnetic beads (MB)-based SELEX is a commonly used method for screening of aptamers. Herein, three types of MB-based SELEX with different immobilization manners were carried out to systematically study and compare the screening efficiency, which may provide reference for screening of aptamers specific for low-molecular targets. After screening and sequencing, the obtained aptamer sequences were aligned and analyzed, among which three aptamers were selected as candidates. Then, the aptamer ap 3-1 with the highest affinity was optimized according to the results of molecular simulation docking. An optimal aptamer ap 3-1-a with 21 nucleotides (nt) was eventually obtained, with the K d value of 55.71 ± 12.29 nM. To verify the practicability of the aptamer, a fluorescent biosensor for Neu5Ac was constructed employing graphene oxide (GO)-based fluorescence quenching and Neu5Ac-induced fluorescence restoration. The biosensor showed a linear response range within 20−1000 nM. The specificity was verified and the interferents-resistance was validated by detecting Neu5Ac in actual samples. This study may inspire the efficient and universal strategy for screening and application of robust and practicable aptamers for low-molecular targets.
研究领域:
生物化学
·
分子生物学
·
分析化学
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
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.
研究领域:
生物化学
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分子生物学
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医学诊断
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Linlin Wang,Shaopu Liu,Chenxia Hao,Xiaolong Zhang,Chaoqun Wang,Youqiu He
A dual-fluorescence “turn off–on” biosensor, which consists of quantum dots (GSH-CdTe QDs) whose fluorescence was quenched by safranine T (ST) via an electron transfer process, had been developed for herring sperm DNA (hsDNA) detection. Initially, in the “turn off” stage, the strong fluorescence of GSH-CdTe QDs could be effectively quenched by ST owing to the occurrence of the electron transfer from the photoexcited GSH-CdTe QDs to ST. And then, the high affinity of DNA to ST enabled the ST attached to the surface of GSH-CdTe QDs, to become embedded into hsDNA double helix structure to form stable complex and moved away from the QDs. Therefore, the recognition of hsDNA could be realized via the fluorescence restoration of the QDs–ST based biosensor, namely of the fluorescence “turn on” procedure. This designed biosensor exhibited good sensitivity and selectivity, for the reason that the detection limit for DNA reached 10.8 ng/mL, meanwhile, neither biologically relevant metal ions, common organic compounds, nor the amino acid had any significant interference in the detection mode. Hence, this simple, fast, sensitive, and selective biosensor owned perfect analysis applications in biochemical DNA monitoring.
研究领域:
生物化学
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分子生物学
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
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2024
·Vol. 191
·期 8
·1-9
·
IF 5.3
Chen Jinri,Xu Dong,Huang Qiang,Wang Shujun,Li Fuhou,Wu Shaojie,Wang Weixia,Zhou Nandi
Sialyl-Lewis x (SLe x ) is a tetrasugar, which plays an important role in initial inflammation and cancer cell metastasis, and can be used as a marker for cancer diagnosis and prognosis or a therapeutic target. Detecting SLe x from complex biological media remains a significant challenge. Herein, a single-stranded DNA aptamer of SLe x was screened based on the double-stranded DNA library–modified magnetic bead (MB)-SELEX technology. After 14 rounds of screening, 12,639 sequences were obtained and divided into nine families. Three representative sequences were selected based on the number of sequence repeats and Gibbs binding free energy, and the aptamer SLe x -Apt2 with 80 nt length ( K d = 23.01 nM) had the best affinity and relatively high specificity for targeting SLe x . Then, a novel dual-recognition fluorescent biosensor for SLe x -sensitive detection based on aptamer SLe x -Apt2 bio-dots and 3-aminobenzoboric acid–modified MB was developed. This method can detect SLe x as low as 32 μM and has a good linear response in the range 100 μM to 2 mM. It has the advantages of low preparation cost, good targeting, and avoiding the occurrence of false-positive and false-negative detection results, which makes the biosensor more valuable in biological detection and clinical diagnosis. Graphical Abstract
研究领域:
生物化学
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分子生物学
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临床医学
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DOI
PubMed
高影响因子
Food Science & Nutrition
·
2021
·Vol. 9
·期 7
·3873-3884
·
IF 2.863
Mingcan Qin,Xiaoyi Ma,Shihui Fan,Hangjie Wu,Wanli Yan,Xiaopeng Tian,Jing Lu,Mingsheng Lyu,Shujun Wang
In the present study, a DNAzyme was screened in vitro through the use of a DNA library and crude extracellular mixture (CEM) of Pseudomonas aeruginosa . Following eight rounds of selection, a DNAzyme termed PAE-1 was obtained, which displayed high rates of cleavage with strong specificity. A fluorescent biosensor was designed for the detection of P. aeruginosa in combination with the DNAzyme. A detection limit as low as 1.2 cfu/ml was observed. Using proteases and filtration, it was determined that the target was a protein with a molecular weight of 10 kDa–50 kDa. The DNAzyme was combined with a polystyrene board to construct a simple indicator plate sensor which produced a color that identified the target within 10 min. The results were reliable when tap water and food samples were tested. The present study provides a novel experimental strategy for the development of sensors based on a DNAzyme to rapidly detect P. aeruginosa in the field.
研究领域:
分子生物学
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微生物学
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生物传感技术
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Shihua Luo,Lifeng Zhang,Xiaohe Zhang,Jiezhen Pan,Mei Wang,Ye Zhang,Aiping Qin,Wencheng Chen
The integrity of sperm DNA is essential for proper fertilization and embryonic development. Various methods exist to evaluate sperm DNA fragmentation. However, they are frequently hindered by high costs, complex procedures, and low precision. Herein, a simple and sensitive fluorescence biosensor was developed using the terminal deoxynucleotidyl transferase (TdT) and CRISPR/Cas12a system for analyzing sperm DNA fragmentation. This biosensor efficiently measures the number of 3′-OH, which correspond to the mean number of sperm DNA breakpoints (MDB). In the presence of TdT, the dATP nucleotide polymerized at the 3′-OH end of the primer, generating numerous polyA-tails, which were detected by CRISPR-guided RNA (crRNA) at the same time. Ultimately, the trans -cleavage activity of CRISPR/Cas12a was activated, which then cleaved the spherical nucleic acids (SNA) reporter, generating amplified fluorescence signal. The proposed method exhibited a broad linear detection for the quantity of 3′-OH ends in single-stranded DNA, from 1 fM to 10 pM, with a limit of detection of 2.5 fM. Moreover, the platform was successfully validated with clinical sperm samples and demonstrated a stronger correlation with sperm motility compared to traditional DNA fragmentation index (DFI) test. Therefore, the developed method offers advantages such as ease of use, elevated sensitivity, and mimimal instrumentation requirements, making it suitable for clinical application.
研究领域:
生殖生物学
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分子诊断学
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基因编辑技术
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产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2022
·Vol. 46
·期 36
·17479-17486
·
IF 3.925
Zhenzhen Cui,Liang Guo,Zhenyu Jin,Lele Ma,Huaixia Yang,Mingsan Miao
Ochratoxin A (OTA) seriously affects the quality of herbal medicines and threatens the health of patients. Therefore, accurate detection of OTA in herbs is of great significance. Here, we fabricated a signal-off fluorescent biosensor for the highly sensitive detection of OTA by the activator regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) signal amplification technique. First, the specific aptamer sequence was immobilized on carboxylic MBs by an amide reaction. At the same time, the initiator was attached to the cDNA. Then the aptamer and cDNA formed a double helix structure through complementary base pairing. Afterwards, numerous fluorescent monomers were grafted onto the cDNA by ARGET ATRP, which emitted strong fluorescence. Upon addition of OTA, it could be specifically recognized by the aptamer and displace the cDNA, generating fluorescence quenching. Under optimal conditions, the detection limit of the as-prepared sensor was 7.6 fg mL−1 with a detection range of 2 × 10−3 ng mL−1 to 2 × 103 ng mL−1. Furthermore, this sensor exhibited high selectivity and reproducibility, and was successfully applied to the detection of OTA in samples of forsythia, astragalus, licorice, xanthium and malt. The outstanding performance of this biosensor suggests its possible application in the detection of mycotoxin in herbal medicine samples.
研究领域:
分析化学
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毒理学
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药理学
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产品关联
DOI
高影响因子
Analytical Methods
·
2023
·Vol. 15
·期 44
·6015-6020
·
IF 3.1
Shanshan Liu,Nian Wang,Li Li,Yi Liu
Diseases such as liver cancer, extrahepatic biliary obstruction and osteocarcinoma are closely associated with the abnormal level of alkaline phosphatase (ALP). Hence, it is essential to develop a convenient assay to detect ALP activity. Herein, a novel signal-on fluorescent biosensor on account of the fluorescence signal of the aggregation-induced emission (AIE) fluorochrome 2,2′,2′′,2′′′-((ethene-1,1,2,2-tetrayltetrakis(benzene-4,1-diyl))tetrakis(oxy))tetraacetic acid (EBTAC) encapsulated zeolitic imidazolate framework-8 (ZIF-8@EBTAC) was designed to monitor ALP. Due to the aggregation-induced emission of EBTAC, the synthetic ZIF-8@EBTAC shows robust fluorescence. Once pyrophosphate (ppi) was added, its complexation with Zn2+ in ZIF-8 triggered the collapse of the ZIF-8 framework, releasing encapsulated EBTAC molecules and restoring to free state, leading to the dramatical decrease in fluorescence. ALP could catalyze the hydrolysis of ppi to phosphate (pi), which is difficult to bind to Zn2+ and has little effect on the fluorescence of ZIF-8@EBTAC. Therefore, with the assistance of the substrate ppi, the ultimate fluorescence of ZIF-8@EBTAC was positively related with ALP activity. The constructed biosensor was able to monitor the ALP activity well from 0.01 to 100 U L−1, and a detection limit of 0.01 U L−1 was achieved. Based on the ability of EBTAC serving as a fluorescent probe with aggregation-induced luminescence properties, this proposed design can be applied to diverse targets and provide new ideas for the establishment of fluorescent biosensors.
研究领域:
生物化学
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医学诊断
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材料科学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）