ANALYTICAL BIOCHEMISTRY
·
2018
·
IF 2.275
Lide Gu,Wanli Yan,Shu Liu,Wei Ren,Mingsheng Lyu,Shujun Wang
A novel screening method for protein aptamer selection was developed in this study. Aptamers with high affinity and specificity to the surface recombinant antigen of Helicobacter pylori (HP-Ag) and to tumor markers carcinoembryonic antigen (CEA), cancer antigen 125 (CA125) and cancer antigen 19-9(CA19-9) were screened using trypsin enhanced screening method. Briefly, the target proteins above were immobilized onto 96-well polystyrene plates and incubated with a single-stranded DNA (ssDNA) library for aptamer selection. Then, trypsin was introduced to digest the proteins and obtain ssDNA that bound to the target proteins with high specificity. The concentration of ssDNA that shed from protein-ssDNA complexes was detected. After sequencing, the enrichment of target-specific aptamers was monitored and the affinity of each aptamer was analyzed. Urea, which has been reported in other article, was used to compare with trypsin. The results revealed that trypsin was more effective than urea for protein aptamer selection. The protocol used in this study provided a novel method for generating aptamers.
研究领域:
分子生物学
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生物技术
·
蛋白质工程
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DOI
PubMed
高影响因子
PHYSICAL CHEMISTRY CHEMICAL PHYSICS
·
2021
·Vol. 23
·期 46
·26356-26365
·
IF 3.676
Dongliang Li,Rong Chen,Xun Zhu,Dingding Ye,Yang Yang,Wei Li,Haonan Li,Yijing Yang,Qiang Liao
In this study, a contactless, flexible, and interference-free light fueled method has been developed to enhance the mixing between the ssDNA and dynabeads in a droplet, which enables rapid probe preparation for promoting the probe technology based on open surface droplet microfluidics. In this light fueled method, the use of the photothermal effect of a focused infrared laser can easily create non-uniform temperature distribution and accordingly the surface tension gradient over the interface as a result of the localized heating effect, which thereby initiates the Marangoni flow in a droplet. Experimental results confirm that the light-induced Marangoni flow greatly enhances the mixing, ensuring rapid and efficient binding between the ssDNA and dynabeads. Moreover, the mixing intensity and degree can be simply tuned by controlling the laser intensity and laser heating time. The light fueled rapid mixing method developed in the present study paves the way for rapid bio-chemical detection.
研究领域:
微流控技术
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生物化学
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光学工程
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PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Dawei Li,Shen Ling,Hongsong Wu,Zhaoqi Yang,Bei Lv
In addition to its application in genome editing, the CRISPR/Cas system provides a new molecular tool for the construction of biosensor platforms due to its high target sequence specificity and programmability . To exploit the potential of the CRISPR/Cas system for detecting non-nucleic-acid targets, two ultrasensitive biosensors (sensor-ss and sensor-ds) consisting of allosteric aptamer probes integrated with the CRISPR/Cas12a system were designed and constructed for tobramycin detection. Both sensors utilized hairpin DNA containing a tobramycin aptamer sequence as the target recognition probe, which can respond to target binding and further produce signal transduction sequences. However, there are two differences between the sensors. First, in sensor-ss, single-stranded DNA was set to be the activator to trigger cutting activity ( trans -cleavage) of CRISPR/Cas12a, leading to cleavage of the FQ-reporter (a short single-stranded DNA labeled with a fluorophore and quencher at each end) and increasing the fluorescent signal. However, double-stranded DNA containing a protospacer adjacent motif (PAM) site was produced in sensor-ds as the signal transduction sequence, which was recognized by CRISPR/Cas12a, resulting in the cleavage of the FQ-reporter. Second, sensor-ss involved a strand displacement amplification (SDA) reaction as the signal enhancement step, while no DNA amplification procedure was designed in sensor-ds. Through DNA sequence and reaction condition optimization, sensor-ds showed a linear relationship between the fluorescence response and tobramycin concentrations ranging from 10 to 300 pM with a limit of detection (LOD) as low as 3.719 pM. Although sensor-ss exhibited higher sensitivity (LOD = 1.542 pM) than sensor-ds, rapidly reaching the fluorescent signal saturation platform led to a narrow linear response range (5–30 pM). Moreover, our CRISPR/Cas12a-based assay can realize on-site detection by the naked eye readout under UV light. The constructed sensor-ds was also used to detect tobramycin in milk and lake water samples, and the results indicated that the detection system was reliable. The design strategy reported in this study could shed new light on CRISPR/Cas12a-based biosensor construction for small molecule detection.
研究领域:
分子生物学
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分析化学
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医学检测
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DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Yingzhou Tao,Fang Luo,Longhua Guo,Bin Qiu,Zhenyu Lin
Colorimetric platform using the aggregation of gold nanoparticles (AuNPs) is a pretty simple method for biosensing, but advanced instruments such as specterophotometer is still needed to achieve accurately quantitative readout. Aggregated AuNPs exhibit excellent photothermal properties under near-infrared laser irradiation, which is significantly different from non-aggregated AuNPs. Herein, given the different photothermal effect, we translated the AuNPs-based colorimetric assay into a photothermal assay for the quantitative detection of adenosine using a thermometer as readout. Short single-stranded DNA (ssDNA, adenosine aptamer) was adsorbed on the surface of AuNPs and hence prevented the aggregation of AuNPs under high ionic concentration. The presence of adenosine caused the structural change of ssDNA and the AuNPs became aggregated. The enhanced temperature under NIR-laser irradiation has a linear response to the concentration of adenosine in the range of 2.0–50.0 μM. The detection limit was 1.7 μM. This proposed method is portable, easy and applicable to the quantitative assay of other targets by simply replacing of the sequence of ssDNA.
研究领域:
生物传感
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纳米技术
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DOI
PubMed
高影响因子
JOURNAL OF RADIOANALYTICAL AND NUCLEAR CHEMISTRY
·
2018
·Vol. 319
·期 1
·69-73
·
IF 1.181
Duan Xiaoyan,Li Jianbo,Lu Xiaoling
In the field of nanotechnology, DNA-based nanoscale materials have facilitated the construction of DNA polyhedrons with different shapes and sizes by using predictable base pairing and highly tunable conformation. In this study DNA bipyramid nanostructures with one arm chain (T20-DBNs) were successfully prepared in a single annealing procedure. 99m Tc-A20 ssDNA was obtained by radiolabeled DTPA-A20 with 99m Tc, and then 99m Tc-DTPA-DBNs were obtained by hybridizing T20-DBNs with 99m Tc-A20 ssDNA. We focused on studying a method of 99m Tc radiolabeling DNA nanostructures with DTPA as a chelate, and hoped to develop a new SPECT molecular imaging probe based on DNA nanostructures.
研究领域:
纳米技术
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放射医学
·
分子影像学
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DOI
高影响因子
ACS Applied Polymer Materials
·
2022
·
IF 4.855
Shanjin Hu,Xiaoxue Du,Yanhui Bi,Ping-Ping He,Yali Mu,Chang Liu,Qi Gao,Mengyuan Yin,Weiwei Guo
As smart soft materials with programmable functions and excellent biocompatibility, DNA hydrogels have great potential in a variety of application fields, such as in the fabrication of soft biosensing devices. However, most DNA hydrogels are formed by self-assembly of multibranched DNA structures or with the assistance of enzymatic processes or chemical modifications, which significantly increases the complexity of material designing and the cost of construction. Herein, we report on the construction of responsive DNA hydrogel based on the self-assembly of one short single-stranded DNA (ssDNA), and the self-assembly can also occur within a polymeric matrix to construct interpenetrating network (IPN) hydrogel. Mechanism investigation reveals that the self-assembly between the short ssDNA strands leads to the formation of long linear DNA structures and looped structures, and the entanglement between these DNA assemblies at high concentrations results in the formation of three-dimensional hydrogel networks. By programming the sequence of the ssDNA strand, pH-responsive DNA hydrogel or catalytic DNA hydrogel can be constructed. Furthermore, the interpenetration of the catalytic DNA networks within a polyacrylamide hydrogel matrix leads to the formation an IPN hydrogel with higher structure stability and with excellent catalytic activity on the polymerization of aniline into conductive polyaniline (PANI). The successful construction of conductive PNAI patterns templated by the IPN hydrogel films is further achieved, which may hold potential in soft electronic and biosensing applications.
研究领域:
生物材料学
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分子生物学
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软物质科学
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Chunying Li,Chunying Wei
We have constructed a label-free fluorescent probe using ssDNA-templated silver nanoclusters (Ag NCs), and its quantum yield (QY) was significantly higher (∼29%) compared to traditional water-soluble fluorescent metal nanoclusters. The proposed approach is simple, non-toxic, sensitive, and selective for the detection of mercuric ion (Hg 2+ ). The fluorescence intensity of ssDNA-Ag NCs was quenched significantly in the presence of Hg 2+ , which might be attributed to the strong interactions between them including the dynamic and static quenching mechanisms. Though Ag + induced a moderate interference to Hg 2+ , there was a negligible quenching in the presence of other metal cations. In addition, Hg 2+ could be detected as low as 4.5 nM, within a good linear range from 0 to 150 nM ( R 2 = 0.995). More importantly, this method has also been successfully applied to the detection of Hg 2+ in the real water samples. Thus, the ssDNA-Ag NCs detection system is potentially suitable for Hg 2+ sensing in environmental samples.
研究领域:
生物化学
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环境科学
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分析化学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2017
·
IF 3.154
Nuo Duan,Wenhui Gong,Shijia Wu,Zhouping Wang
Clenbuterol hydrochloride (CLB) is often abused as additive feed for livestock to decrease adipose tissue deposition and to increase growth rate. It raises a potential risk to human health through the consumption of animal product. In this study, aptamers with higher affinity and specificity were screened through 16 selection rounds based on the ssDNA library immobilized systematic evolution of ligands by exponential enrichment (SELEX) technique. After cloning and sequencing, five aptamer candidates were picked out for affinity and specificity assays based on a graphene oxide (GO) adsorption method. The results showed that the aptamer CLB-2 binds specifically against CLB with a dissociation constant, Kd, value of 76.61 ± 12.70 nM. In addition, an aptamer-based fluorescence bioassay was established for CLB analysis. The correlation between the CLB concentration and fluorescent signal was found to be linear within the range of 0.10 to 50 ng/mL with a limit of detection of 0.07 ng/mL. It has been further applied for the determination of CLB in pork samples, showing its great potential for sensitive analysis in food safety control.
研究领域:
食品安全
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分子生物学
·
分析化学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Chan Song,Guan-Yao Wang,De-Ming Kong
This work investigated the interactions of α-Fe 2 O 3 nanoparticles (NPs) with different structural nucleic acids and their fluorescence quenching ability towards fluorophore-labelled nucleic acid probes . Different from bulk α-Fe 2 O 3 samples, nanoscale α-Fe 2 O 3 particles exhibit the unique properties of strong adsorption and fluorescence quenching to fluorophore-labelled single-stranded DNA (ssDNA) probes. Based on these findings, a facile fluorescence method was developed for versatile quantification of nucleic acids. The size scale of NPs makes a significant impact on this sensing platform. Better selectivity was given by bigger NP (50–100 nm)-based nucleic acid-sensing platform compared with smaller NP (30 nm)-based one. In the 50–100 nm α-Fe 2 O 3 NP-based sensing platform, single nucleotide mismatch or single base-pair mismatch can even be effectively discriminated. The targets of micro-RNA (miRNA), ssDNA and double-stranded DNA (dsDNA) are sensitively detected with detection limits of 0.8 nM, 1.1 nM and 0.64 nM ( S / N =3), respectively. Significantly, α-Fe 2 O 3 NPs possess different affinities towards ssDNA probes with different lengths, and can be used as a universal quencher for ssDNA probes labelled with different fluorescent dyes . On the basis of these properties, the pristine α-Fe 2 O 3 NPs hold the potential to be widely utilized in the development of novel biosensors with signal amplification or simultaneous multiple target detection strategies.
研究领域:
生物化学
·
分子生物学
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分析化学
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DOI
PubMed
高影响因子
APPLIED SPECTROSCOPY
·
2023
·
IF 3.5
Ning-Ning Wu,Lin-Ge Chen,Hai-Bo Wang
A sensitive fluorescent sensor has been developed for the determination of tetracycline (TC) using adenine thymine (AT)-rich single-stranded DNA (ssDNA) templated copper nanoclusters (CuNCs) as a fluorescent probe. Fluorescent ssDNA-CuNCs were synthesized by employing AT-rich ssDNA as templates and ascorbic acid as reducing agents through a facile one-step method. The as-prepared ssDNA-CuNCs exhibited strong fluorescence with a large Stokes shift (240 nm) and stable fluorescence emission. In the presence of TC, the fluorescent intensity of ssDNA-CuNCs was obviously decreased through the inner filter effect, due to the spectral overlapping between ssDNA-CuNCs and TC. Under the optimal conditions, the strategy exhibited sensitive detection of TC with a linear range from 2 nM to 30 μM and with a limit of detection of 0.5 nM. Furthermore, the sensor was successfully applied for the detection of TC in milk samples. Therefore, it provided a simple, rapid, and label-free fluorescent method for TC detection.
研究领域:
分析化学
·
生物传感
·
环境监测
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED POLYMER SCIENCE
·
2018
·Vol. 136
·期 8
·47102
·
IF 1.901
B.-Q. Huang,H. Ding,Z.-L. Xu,Y.-J. Tang
In this work, the biomacromolecule, single-stranded deoxyribonucleic acid (ssDNA) was innovatively incorporated into the polyamide layer to tailor the permeate flux and antifouling performance of the nanofiltration (NF) membranes. With active amines groups, the ssDNA was as the aqueous phase monomers along with piperazine (PIP), and reacted with trimesoyl chloride on polyethersulfone substrate to fabricate thin-film composite (TFC) NF membranes. The NF membrane prepared under optimal ratio of ssDNA/PIP had a pure water permeability of 75.8 L m −2 h −1 (improved 58% compared to PIP NF membrane) and Na 2 SO 4 rejection of 98.0% at 6.0 bar. The rejections for different inorganic salts were the order: Na 2 SO 4 (98.0%) > MgSO 4 (89.2%) > MgCl 2 (72.8%) > NaCl (23.0%). Furthermore, the TFC NF membranes showed good antifouling performance in long-term running with 300 ppm bovine serum albumin and humic acid solution. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135 , 47102.
研究领域:
材料科学
·
环境工程
·
生物化学
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DOI
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 2
·1-9
·
IF 5.833
Zhang Dongwei,Liu Yang,Ding Jina,Hayat Kashif,Zhan Xuejia,Zhou Pei,Zhang Dan
A short ssDNA (Apt-21) rationally truncated from the parent 100 nt As(III) aptamer was used for colorimetric determination of As(III). Apt-21 serves dual functions, i.e., recognition of trace As(III) and regulation of AuNPs dispersion by surface attachment, while gold nanoparticles (AuNPs) functioned as colorimetric signal reporters. Under the optimal conditions, the ratio of the absorbance at 650 nm to 520 nm (A 650 /A 520 ) of AuNPs changed proportionally with increasing concentration of As(III), which showed a linear relationship within the concentration ranges 1–30 ppb and 30–100 ppb with a detection limit of 0.18 ppb. The feasibility of this assay was demonstrated by determining As(III) in spiked water samples with mean recoveries ranging from 96.5–107.1%. Graphical abstract
Schematic representation of colorimetric detection of As(III) based on the short ssDNA (Apt-21) and gold nanoparticles (AuNPs)
研究领域:
分析化学
·
环境科学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
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