LUMINESCENCE
·
2022
·Vol. 37
·期 6
·922-929
·
IF 2.613
Tingting Feng,Shuzhu Yan,Zheng Wang,Xiaohui Fan
In this study, we present a facile and low-cost approach for detecting protein kinase A (PKA) by assembling a purpose-designed carboxyfluorescein (FAM)-labelled peptide with carboxylic carbon nanoparticles (CNPs). Fluorescence of the FAM-labelled peptide gradually decreases to a low background signal as a result of the electron transfer from CNPs to FAM-labelled peptide via the peptide, which acts as a bridge. The reaction in the sensor in the presence of adenosine 5′-triphosphate and PKA phosphorylates the substrate peptide and disrupts the electrostatic repulsive force between the CNPs and the peptide, therefore altering the spectroscopic signal of the system. The change in fluorescence signal was directly proportional to the PKA concentration in the range 0–1.8 U/ml with a detection limit of 0.04 U/ml. These results suggest that PKA activity can be effectively measured using the developed PKA biosensor. Moreover, the fluorescence biosensor was successfully used in the investigation of PKA in spiked human embryonic kidney (HEK) 293 cells lysates, indicating its potential applications in protein kinase-related biochemical fundamental research.
研究领域:
生物化学
·
分子生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2022
·Vol. 189
·期 8
·1-10
·
IF 6.408
Zhao Tingting,Yan Weizhen,Dong Fengqi,Hu Xinlong,Xu Yanli,Wang Zhenyu,Shen Yating,Wang Wanrong,Zhao Ye,Wei Wenmei
COVID-19 necessitates the development of reliable and convenient diagnostic tools. In this work, a facile 3D-printed smartphone platform was constructed that achieved reliable visual detection of SARS-CoV-2 by eliminating the effect of ambient light and fixing the camera position relative to the sample. The oligonucleotide probe is modified with orange-red-emitting TAMRA working as an internal standard and green-emitting FAM serving as a sensitive sensing agent. Under 365-nm UV excitation, the emission wavelengths of TAMRA and FAM are 580 nm and 518 nm, respectively. When the probes interact with the targets, the green fluorescence gradually restores while the orange-red fluorescence remains stable. Thus, a striking color transition from orange-red to green could be observed by the naked eye. The detection limit of SARS-CoV-2 nucleic acid is 0.23 nM, and the entire process of color change could be completed in 25 min. Furthermore, the RGB value analysis of the sample solution was conducted using a smartphone for reliable and reproducible discrimination of SARS-CoV-2. The proposed smartphone platform might establish a general method for visual detection of SARS-CoV-2 nucleic acid as well as other virus-related diseases. Graphical abstract
研究领域:
生物医学工程
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分子生物学
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诊断工具
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病毒学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Yu Liu,Gaohua Han,Jing Gong,Xin Hua,Qian Zhu,Sisi Zhou,Ling Jiang,Quan Li,Songqin Liu
Early and accurate diagnosis of hepatocellular carcinoma (HCC) is of significant importance for improving the survival rate and quality of life for HCC patients. The combined detection of alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3), namely AFP-L3%, can greatly improve the accuracy of HCC diagnosis compared with AFP detection. Herein, we developed a novel intramolecular fluorescence resonance energy transfer (FRET) strategy for sequential detection of AFP and AFP-specific core fucose to improve the diagnosis accuracy of HCC. Firstly, fluorescence-labeled AFP aptamer (AFP Apt-FAM) was used to specifically recognize all AFP isoforms, and total AFP was quantitatively determined using fluorescence intensity of FAM. Then, 4-((4-(dimethylamino)phenyl)azo)benzoic acid (Dabcyl) labeled lectins (PhoSL-Dabcyl) were used to specifically recognize the core fucose expressed on AFP-L3 that does not bind to other AFP isoforms. The combination of FAM and Dabcyl on the same AFP molecule could generate FRET effect, thereby quenching the fluorescence signal of FAM and quantitatively determining AFP-L3. After that, AFP-L3% was calculated according to the ratio of AFP-L3 to AFP. With this strategy, the concentration of total AFP, AFP-L3 isoform as well as the AFP-L3% were sensitively detected. Detection limits of 0.66 and 0.186 ng/mL were obtained for AFP and AFP-L3 in human serum, respectively. Clinical human serum test results showed that AFP- L3 % test was more accurate than AFP assay to distinguish healthy people, HCC patients and benign liver disease patients. Therefore, the proposed strategy is simple, sensitive and selective, which can improve the accuracy of early diagnosis of HCC, and has good clinical application potential.
研究领域:
医学诊断
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癌症研究
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分子生物学
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DOI
PubMed
高影响因子
Current Analytical Chemistry
·
2024
·Vol. 21
·期 8
·1005-1012
·
IF 1.7
Jiangxiong Zhu,Yang Wang,Lumei Wang,Xueqing Geng,Linnan Yang,Ting Zhao,Yun Deng
ObjectiveThe objective of this study is to develop a novel fluorometric aptasensor employing fluorescence resonance energy transfer (FRET) for the detection of Cadmium (II) (Cd2+) in water and food samples. The constructed aptasensor employed a fluorophore-quencher labeled aptamer combination not previously reported for Cd2+ detection. Additionally, its simple mix-and-detect pattern without immobilization or material-assisted steps represented an innovative design.MethodsUtilizing 6-carboxyfluorescein (FAM)-modified aptamers and maleimide (BHQ-1)-modified aptamer complementary chain to construct a fluorescent detection probe, this aptasensor achieved a rapid, sensitive, and selective detection of Cd2+. Without Cd2+, the aptamer and its complementary strand undergo base pairing, bringing the FAM closer to the BHQ-1, and leading to FRET and a subsequent decrease in fluorescence intensity. The introduction of Cd2+ preferentially brought to the aptamer, changing its conformation and preventing the quenching of FAM by BHQ-1, thereby restoring the fluorescence intensity of the aptasensor.ResultsFollowing optimization of experimental parameters, the aptasensor exhibited a linear response to Cd2+ concentrations ranging from 5 to 1200 nM, with a detection limit (LOD) of 0.43 nM. The aptasensor’s performance was unaffected by the presence of various ions, indicating its high specificity. Moreover, it could rapidly and accurately detect Cd2+ in water and food samples, including tap water, lake water, grapes, cabbage, and broccoli, demonstrating its substantial potential for practical application.ConclusionTherefore, the developed aptasensor represents an important tool for effective Cd2+ detection in water and food matrices, highlighting its potential as a critical tool for environmental monitoring and food safety.
研究领域:
环境监测
·
食品安全
·
分析化学
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生物传感器
·
重金属检测
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2013
·
IF 3.535
Ting Wen,Nian Bing Li,Hong Qun Luo
A novel sensing system has been designed for the Cu 2+ detection based on the quenched fluorescence signal of 6-carboxyfluorescein (FAM)-labeled DNA/polyethyleneimine. The FAM-labeled DNA showed weak fluorescence at the emission wavelength of 530 nm in a pH 4.8 acetic acid-sodium acetate buffer solution. When the polyethyleneimine was introduced to the FAM-labeled DNA solution, the fluorescence signal was greatly amplified. Subsequently, upon the addition of Cu 2+ , Cu 2+ can be captured by the amino groups of the polyethyleneimine to form an absorbent complex, resulting in a strong quenching of the fluorescence of FAM-labeled DNA/polyethyleneimine via a fluorescence resonance energy transfer effect. Herein, we have demonstrated that this facile methodology can offer a rapid, reliable, and selective detection of Cu 2+ with a detection limit as low as 20 nM and a linear range of 100–1600 nM. Furthermore, the detection results for Cu 2+ in a lake water sample obtained by this sensing system agreed well with that by atomic absorption spectroscopy, suggesting the potential application of this sensing system.
研究领域:
分析化学
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环境科学
·
材料科学
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产品关联
DOI
高影响因子
JOURNAL OF NANOPARTICLE RESEARCH
·
2022
·Vol. 24
·期 5
·1-10
·
IF 2.533
Chen Junjie,Li Xiangqian,Yu Xiaoqi,Zhou Wei,Wang Qianming
Two-dimensional platforms generate considerable interests due to diverse application. We report a “turn-on” fluorescent nanoprobe for prostate-specific antigen (PSA) detection. It is based on cobalt oxyhydroxide nanosheets (CoOOH NSs). Fluorescein amidite (abbreviated as FAM)-labeled aptamer probe (abbreviated as FAM-aptamer) has been adsorbed onto the nanosheets. Energy transfer between substrate and optical species has induced the quenching of FAM-aptamer emission. The strong affinity of FAM-aptamer to the target PSA causes the formation of a rigid aptamer structure. The integration between FAM-aptamer and CoOOH NSs has been drastically affected. The release of the aptamer probe from the nanosheet surface has been realized. The green luminescence has been recovered. The dynamic nanosensor exhibits highly sensitive performance toward PSA with a linear detection range from 0.1 to 5 ng/mL. Its detection limit is measured to be 56.1 pg/mL. Therefore, a simple and efficient sensing platform for the detection of prostate cancer can be established.
研究领域:
生物纳米技术
·
分析化学
·
癌症诊断
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2017
·
IF 6.32
Zhen Zhang,Yuting Jiao,Mengting Zhu,Shusheng Zhang
Here, we propose a strategy for unique nuclear-shell biopolymers initiated by telomere elongation for telomerase activity detection and precise drug delivery to individual cancer cells. Telomerase-triggered DNA rolling-circle amplification (RCA) is used to assemble nuclear-shell biopolymers with signal molecules for selective cancer cell recognition and efficient drug delivery to targeted individual cells. This strategy not only should allow the creation of clustered 5-carboxyfluorescein (FAM)-fluorescence spots in response to human-telomerase activity in individual cancer cells but also could efficiently deliver drugs to reduce the undesired death of healthy cells. These findings offer new opportunities to improve the efficacy of cancer cell imaging and therapy.
研究领域:
癌症生物学
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分子生物学
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生物医学工程
·
药学
✓ 已验证
DOI
PubMed
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2023
·
IF 1.2
Peng WANG,Wen-Heng LI,Xiao-Hong WANG,Dong-Shan XIANG,Kun ZHAI
We developed a new quantitative detection method for trace sulfur ions (S 2− ) in water using a Cu-containing metal–organic framework material (CuBDC) and 6-carboxyfluorescein group (FAM)-labeled DNA of arbitrary sequence. Without S 2− in the system, the FAM-labeled DNA can be adsorbed on the CuBDC surface through π–π stacking, hydrogen bonding, and van der Waals forces, and then the fluorescent dye FAM is near the Cu 2+ on the CuBDC surface, and the fluorescence of FAM is quenched by a photoinduced electron transfer mechanism between Cu 2+ and FAM. With S 2− in the system, S 2− reacts with Cu 2+ in CuBDC to form the more structurally stable compound CuS, leading to CuBDC decomposition, and the FAM-labeled DNA detaches from the CuBDC surface and the fluorescence of FAM recovers. The quantitative detection of S 2− is realized according to the change in FAM fluorescence intensity. Under optimized conditions, a good linear relationship holds between the fluorescence intensity (F i ) of FAM and the concentration ( C i ) of S 2− in the range of 2–120 nmol/L, and the detection limit is 1.5 nmol/L (3 σ, n = 9). The method has good selectivity and accuracy, and it can be applied to rapidly and accurately analyze S 2− in real water samples.
研究领域:
分析化学
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环境科学
·
材料科学
·
生物化学
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产品关联
DOI
高影响因子
iScience
·
2022
·
IF 6.107
Junzhe Zhang,Wenxue Lei,Yixuan Meng,Chengqian Zhou,Baihong Zhang,Jun Yuan,Min Wang,Dongqing Xu,Xiangchun Meng,Wenli Chen
Summary Huanglongbing ( HLB ) is a devastating disease of citrus, which is mostly caused by Candidatus Liberibacter asiaticus (C Las ). To realize the specific application of nano-transgenic technology in HLB, AuNPs-PEI (Gold Nanoparticles-Polyethylenimine) was used to carry foreign genes into the leaves of periwinkle ( Catharanthus roseus ) by infiltration. Here, we demonstrated that NPR1-GFP protein expression was observed from the 12th hour to the 10th day after infiltrating AuNPs-PEI-pNPR1 ( Arabidopsis thaliana nonexpressor of pathogenesis-related gene 1 )-GFP. Fluorescence of mCherry was observed 6 h after AuNPs-PEI-pNLS (nuclear localization signal sequence)-mCherry infiltration and fluorescence of FAM was observed in the nucleus 4 h after AuNPs-PEI-FAM-siRNA NPR1 infiltration. In addition, NPR1-GFP expression in C Las-infected periwinkle leaves was significantly higher than that in healthy periwinkle leaves after infiltration. Our work confirmed that the expression of exogenous NPR1-GFP could reduce the C Las titers by promoting the expression of PR ( pathogenesis related ) genes and ICS ( isochorismate synthase ) gene.
研究领域:
植物学
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遗传学
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分子生物学
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植物病理学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2020
·Vol. 412
·期 23
·5627-5635
·
IF 3.637
Tan Hongxia,Guo Ting,Zhou Hongyuan,Dai Hongjie,Yu Yong,Zhu Hankun,Wang Hongxia,Fu Yu,Zhang Yuhao,Ma Liang
Zearalenone (ZEN) is a type of estrogenic mycotoxin commonly occurring in cereals. The aim of this study was to design a simple, rapid, inexpensive and ultrasensitive fluorescence assay for the determination of ZEN. Here, amino-modified mesoporous silica nanoparticles (MSNs-NH 2 ) were synthesized to be the positive charge-rich reactor. A 6-carboxy-fluorescein-labeled aptamer (aptamer-FAM) was designed as the signal probe, ZEN-capture probe and negative charge reactor. In the absence of ZEN, the negatively charged aptamer-FAM combined with the positively charged MSNs-NH 2 in an electrostatic manner. In the presence of ZEN, the fluorescence intensity in the supernatant increased significantly because the aptamer-FAM could bind to ZEN instead of MSNs-NH 2 . Under the optimal experimental conditions, this assay exhibited excellent specificity, repeatability and a wide linearity range of 0.005–150 ng/mL, with a detection limit of 0.012 ng/mL. Additionally, it showed high recovery (83.3–101.5%) for the spiked samples. There was no statistically significant difference in the ZEN concentrations detected by the proposed assay and HPLC in naturally contaminated samples. Overall, this design provides a new strategy for the rapid, inexpensive and sensitive detection of ZEN, and it could be applied to develop fluorometric assays for different targets by the selection of appropriate aptamers.
Graphical abstract
研究领域:
食品科学
·
分析化学
·
纳米技术
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2023
·Vol. 47
·期 28
·13228-13234
·
IF 3.3
Yuanyi Wu,Shihan Liu,Yumei Xiao,Qian Li,Qin Xiang,Xianqin Song,Shanglin Wu,Kaiyong Wang,Yi Yang
Accurate detection of DNA is critical for biomedicine and food safety. Layered double hydroxides (LDHs) are a class of two-dimensional layered materials that have received extensive attention in the fields of sensors, catalysis, and materials science. The application of LDHs in DNA sensing in turn represents a fascinating topic. In this study, we found that both 6-carboxyfluorescein (FAM)-modified single-stranded DNA (ssDNA) probes and probe/target double-stranded DNA (dsDNA) duplexes could be adsorbed by LDHs, which resulted in high fluorescence quenching (>95%) for the first time. Based on the extraordinarily high quenching efficiency of LDHs, we developed a fluorescent sensing platform for simple and highly efficient detection of DNA, using Ni/Al-LDHs as fluorescence quenchers. For the selective degradation of dsDNA, exonuclease III was integrated into the sensor. In the sensing platform, the fluorescence of the probes was rapidly quenched by Ni/Al-LDHs. In the presence of the target DNA, dsDNA was formed, which was subsequently selectively degraded by exonuclease III into short fragments, resulting in the maintenance of the fluorescence of FAM labels. The linear range was 10–400 nM. The detection limit was 1.38 nM. This sensor accomplished the detection of target DNA within 50 min. It provides a convenient, rapid and versatile sensing platform for DNA biomarkers.
研究领域:
生物医学工程
·
材料科学
·
分析化学
·
生物传感器
✓ 已验证
DOI
高影响因子
FOOD CHEMISTRY
·
2022
·
IF 9.231
Pengfei Ma,Hua Ye,Hualin Guo,Xiaoyuan Ma,Lin Yue,Zhouping Wang
Herein, for the first time, a novel truncation strategy for aptamers’ sequences based on molecular docking was reported for the first time. T-2 toxin and AFB 1 aptamers were employed as models and fluorescence polarization was used to measure the affinity of aptamers. The T-2 toxin aptamer (T40) with 40 bases and the AFB 1 aptamer with 32 bases were successfully obtained with increased affinities compared with original aptamers. Meanwhile, circular dichroism, FAM-BHQ1 dual labeled T40, enzyme digestion, and molecular dynamics simulation were performed to investigate the binding mechanism between aptamer T40 and T-2 toxin. Finally, based on MnO 2 nanosheet and the fluorescence amplification effect of SYBR Green I, simplified and rapid fluorescence detection of T-2 toxin was achieved with a detection range of 0.03 nM to 30 nM, indicating the great potential of aptamer in practical detection applications.
研究领域:
分析化学
·
分子生物学
·
毒理学
·
纳米技术
✓ 已验证
DOI
PubMed
高影响因子
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