BIOELECTROCHEMISTRY
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2022
·
IF 5.76
Hongmei Yang,Jing Ren,Mei Zhao,Chen Chen,Fang Wang,Zilin Chen
The detection of methylation level in O 6 -methylguanine-DNA methyltransferase (MGMT) gene is of great significance for diagnosis, prognosis and treatment of glioblastoma. Herein, we proposed an electrochemical immunosensor for detection of MGMT gene methylation. Graphene oxide-magnetic nanoparticles-β-cyclodextrin (GO-Fe 3 O 4 -β-CD) nanocomposite was covered on electrode, and then anti-5-methylcytosine (5mC) antibody was immobilized on modified electrode via host–guest interaction with β-CD. The immobilized antibody specifically recognized and directly captured 5mC on DNA containing MGMT promoter sequence (MGMT-DNA). The methylation level in MGMT-DNA could be evaluated using Ru(NH 3 ) 6 3+ as an electrochemical signal indicator. The redox recycling of Ru(NH 3 ) 6 3+ was triggered by Fe(CN) 6 3− and further amplified current response. A detection limit as low as 0.0825 pM was obtained. Finally, this electrochemical method was successfully applied to analysis of methylated MGMT-DNA in biological sample and detection of other methylation modifications, such as N6-methyladenosine in RNA.
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 13
·3755-3763
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IF 4.478
Wu Shujie,Wang Songen,Li Zongbing,Wu Chun,Ma Dik-Lung,Miao Xiangmin
In this work, we synthesized an iridium(III) complex and studied its selective ability to interact with a specific G-quadruplex DNA sequence (GTGGGTAGGGCGGGTTGG). Results showed that the iridium(III) complex exhibits high selectivity for the G-quadruplex DNA and could be used as an efficient electrochemiluminescence (ECL) probe in a switch-on assay format for the detection of double-stranded DNA (dsDNA). To construct the assay, a hairpin-structured capture probe (CP) which was modified by thiol at its 3′ end and contained the G-quadruplex sequence at its 5′ end was firstly immobilized on a gold electrode. Upon the specific recognition of the dsDNA sequence with the corresponding CP, the hairpin structure of the CP was opened to free G-quadruplex sequence, forming the G-quadruplex structure with the assistance of K + . Then, the iridium(III) complex was able to specifically interact with the G-quadruplex to produce an obvious ECL signal that was proportional to the dsDNA concentration. Notably, this iridium(III) complex/G-quadruplex-based strategy was universal and was not limited to the analysis of DNA using specific sequences, thus opening a new avenue for the application of the G-quadruplex-selective iridium(III) complex in the field of ECL.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Min Chen,Zhen Song,Xiqin Yang,Zhiling Song,Xiliang Luo
Herein, a universal strategy for the construction of highly sensitive and low fouling biosensors was proposed based on antifouling peptides conjugated with recognizing DNA probes . The peptide-DNA conjugate was formed through a reagent-free click reaction between a typical DNA aptamer modified with 5′-dibenzocyclooctyne (DBCO) and the designed antifouling peptide terminated with biotin and the azide group at its two ends. With the assistance of streptavidin (SA), the electrochemical biosensor was constructed via immobilization of the straight peptides and peptide-DNA conjugates in sequence onto the electrode surface modified with electrodeposited poly(3,4-ethylenedioxythiophene) (PEDOT) and gold nanoparticles (AuNPs). The prepared biosensor exhibited excellent antifouling performances in various human bodily fluids such as serum, sweat and urine, with a wide linear response range for CA125 from 0.01 U mL −1 to 1000 U mL −1 , and a low limit of detection of 0.003 U mL −1 . Combining the advantages of the antifouling peptide and recognizing DNA probe, this sensing strategy was capable of assaying CA125 in undiluted human serum, and it also offered a highly promising way for the development of different antifouling biosensors through the conjugation of antifouling peptides with various DNA probes.
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ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Jia-Hui Dong,Chen-Yu Xue,Xin-Ying Zhong,Ying-Lin Zhou,Xin-Xiang Zhang
Oxidative DNA damage is tightly linked to the development of multiple age-related diseases. The prominent oxidation product is 8-oxo-7,8-dihydroguanine (OG), which has been proved to be an important epigenetic-like biomarker. Quantification of the locus-specific OG frequency includes quantitative and locating information, which is of great significance for exploring the functional roles of OG in disease induction and gene regulation. Herein, an ultrasensitive quantification of OG at single-base resolution was established using real-time fluorescence quantitative polymerase chain reaction as an amplification tool. Based on the coding property of Bsu DNA polymerase that incorporates adenine on the opposite site of OG and the selectivity of the ligase for perfectly matched sequences, the difference between OG and G on the sequence could be enlarged. Well-performed Taq DNA ligase was selected out, and as low as 46.2 zmol of target DNA with an OG site and an OG frequency of 5% could be detected. G contents on a specific site were also detectable based on the similar principle, thus the OG frequency of this locus could be accurately determined by a standard addition method. This strategy was successfully applied to the evaluation of locus-specific OG in both model DNA and genomic DNA from human cervical carcinoma cell lines under multiple oxidative stress, showing the potential for functional research and dynamic monitoring of critical OG sites.
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高影响因子
Journal of Materials Chemistry B
·
2022
·Vol. 10
·期 34
·6443-6452
·
IF 7.571
Dan Xiu,Sibo Zhao,Zhenhua Li,Yanan Xu,Yao Wang,Zhijun Zhu,Min Zhang,Christopher D. Snow,Laurence A. Belfiore,Jianguo Tang
In this work, a designed porous DNA crystal with high intrinsic biocompatibility was used as the scaffold material to load fluorescent guest molecules to detect anti-cancer drugs. It is shown here that the synthesized crystals have the characteristics consistent with the designed large solvent channels, and can therefore accommodate guest molecules such as fluorescent proteins that cannot be accommodated by less porous crystals. Eu(TTA)3phen and Tb(acac)3phen lanthanide complexes were individually noncovalently loaded into the porous crystals, resulting in hybrid luminescent DNA crystals. Emodin, an anti-cancer, anti-tumor, anti-inflammatory drug, was found to quench lanthanide complexes in solution or in crystals. Notably, emodin is the active ingredient of Lianhua Qingwen Capsule, an anti-COVID-19 drug candidate. Therefore, the porous DNA crystals reported here have potential applications as a biocompatible and theranostic delivery biomaterial for functional macromolecules.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Xin Shang,Jiayang He,Chunli Yang,Siyu Zuo,Ruo Yuan,Wenju Xu
Development of logic AND-gated multivariate assay systems of DNA-scaffolded green- and red-fluorescent Ag nanoclusters ( g- AgNC and r- AgNC) is promising and remains challenging. By using two DNA segments ( T o and T r) as dual-targeting inputs, herein we report an innovative strategy that utilizes targets-recycled amplification to create a sensitive AND-gated light-up ratiometric sensor with reversely changed fluorescence of g- AgNC and r- AgNC. The design is based on that: two hairpins ( H o and H r) are individually recognizable to T o and T r, and two hybridized duplexes are resultantly implemented for transient dissipative base pairing, i.e. H o· H r, releasing T o and T r for repeated recycling. In an original stem-loop hairpin reporter (HR), one exposed overhang is to host g -AgNC. When HR is unfolded by the cooperative sticky toeholds in H o∙ H r, its liberated stem merging with the g -AgNC template is for r- AgNC clustering. Only when meeting simultaneously the overexpressed T o and T r in SARS-CoV-2, the AND-gated ratio fluorescence of r- AgNC over g- AgNC can be lighted-up in a typical assay, enabling highly specific-inputs binding and sensitive dose-dependence down to 0.29 pM. This enzyme-free and label-free AND-logic ratiometirc strategy illustrates the potential of using multicolor AgNCs emitters for more accurate multivariate assay.
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高影响因子
MICROCHEMICAL JOURNAL
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2022
·
IF 5.304
Zenghui Liu,Jinyan Xue,Luyao Chen,Lele Ma,Huaixia Yang,Yaping Zhang,Mingsan Miao
A new, simple and low-cost electrochemical analysis approach employing click polymerization as the signal amplification strategy was developed for the highly sensitive assay of aflatoxin B 1 (AFB 1 ). First, the aptamer served as the capture probe for AFB 1 self-assembled on the surface of the Au electrode via Au-S bonding. After complementary DNA (c-DNA) modified with azide groups was hybridized with the aptamer, a massive click polymer was introduced as a signal tag via click chemistry. Finally, the captured AFB 1 competed with the c-DNA to bind the aptamer and the c-DNA was shed from the electrode surface and the current intensity was reduced. The electrochemical aptamer sensor is easy to operate, selective and sensitive, and also exhibits excellent reliability in detecting AFB 1 in actual samples. A rapid, simple, efficient and sensitive platform for AFB 1 analysis is proposed in this work, and the strategy can be further extended to trace analysis of other analytes, safety detection, environmental monitoring and disease diagnosis after replacing the corresponding capture probes, with promising applications in the fields of food hygiene and safety.
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BIOELECTROCHEMISTRY
·
2022
·
IF 5.76
Wei Zhang,Jimin Yang,Xiaowei Hu
A self-signal electrochemical sensing platform was constructed for direct recognition of circulating tumor DNA (ctDNA) employing black phosphorous nanosheets (BPNS) assembled with flavin adenine dinucleotide (FAD) prepared by ultrasonication approach. FAD provided a highly efficient and stable dispersing medium for acquiring highly dispersed BPNS in aqueous phase. The obtained FAD/BPNS nanocomposite exhibited favorable electrochemical redox activity and was utilized as the interface for the immobilization and hybridization of DNA. The amine-terminated probe ssDNA was covalently assembled onto the FAD/BPNS nanocomposite with plentiful phosphonate groups accompanied by the decrease of the self-signal. The self-redox signal of the nanointerface regenerated after the probe hybridized with the complementary sequence as a result of DNA transformation. Electrochemical response enhanced with complementary DNA concentration from 1.0 × 10 -18 to 1.0 × 10 -8 mol/L with a detection limit of 2.6 × 10 -19 mol/L. The DNA determination platform revealed outstanding sensitivity, specificity and stableness, and was successfully employed in the detection of ctDNA associated with colorectal cancer. The developed biosensing strategy is simple to accomplish and has the potency for the application for diverse morbific gene without sophisticated label procedure.
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APPLIED SURFACE SCIENCE
·
2021
·
IF 6.707
Wenting He,Hao Shen,Zixin Zhou,Ze Huang,Hao Chao,Jiayi Song,Ping Su,Yi Yang
Metal-organic frameworks (MOFs) are excellent platforms for enzyme immobilization. However, strong or weak interactions between enzymes and MOFs often lead to inactivation and enzyme leaching. Herein, single-stranded DNA (ssDNA) with Janus properties was used as a linker between enzymes and the dual carriers of MOFs and graphene oxide (GO), which provided a simple and effective platform to tightly co-immobilize glucose oxidase (GO X ) and horseradish peroxidase (HRP). Mechanistic investigation revealed that the enzyme was immobilized mainly through π-π stacking between nucleobases and GO and coordination of the sugar-phosphate backbone with zirconium clusters in MOFs. This strategy ameliorated traditional surface adsorption issues for enzyme immobilization on MOFs. By introducing GO and ssDNA, the GO X &HRP cascade reaction on MOFs was enhanced significantly, showing better kinetics than the corresponding free GO X &HRP. Furthermore, the immobilized GO X &HRP exhibited substrate selectivity and a wide linear range for glucose detection. This enzyme immobilization strategy should have broad application prospects and practical value in biocatalysis and diagnostics.
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Frontiers in Immunology
·
2021
·
IF 7.561
Yu Zhengqing,Cao Wandi,Gao Xuchen,Aleem Muhammad Tahir,Liu Junlong,Luo Jianxun,Yan Ruofeng,Xu Lixin,Song Xiaokai,Li Xiangrui
Toxoplasma gondii (T. gondii) is an intracellular parasitic protozoan that can cause serious public health problems. However, there is no effectively preventive or therapeutic strategy available for human and animals. In the present study, we developed a DNA vaccine encoding T. gondii oxidoreductase from short-chain dehydrogenase/reductase family (TgSDRO-pVAX1) and then entrapped in chitosan and poly lactic-co-glycolic acid (PLGA) to improve the efficacy. When encapsulated in chitosan (TgSDRO-pVAX1/CS nanospheres) and PLGA (TgSDRO-pVAX1/PLGA nanospheres), adequate plasmids were loaded and released stably. Before animal immunizations, the DNA vaccine was transfected into HEK 293-T cells and examined by western blotting and laser confocal microscopy. Th1/Th2 cellular and humoral immunity was induced in immunized mice, accompanied by modulated secretion of antibodies and cytokines, promoted the maturation and MHC expression of dendritic cells, and enhanced the percentages of CD4+ and CD8+ T lymphocytes. Immunization with TgSDRO-pVAX1/CS and TgSDRO-pVAX1/PLGA nanospheres conferred significant immunity with lower parasite burden in the mice model of acute toxoplasmosis. Furthermore, our results also lent credit to the idea that TgSDRO-pVAX1/CS and TgSDRO-pVAX1/PLGA nanospheres are substitutes for each other. In general, the current study proposed that TgSDRO-pVAX1 with chitosan or PLGA as the delivery vehicle is a promising vaccine candidate against acute toxoplasmosis.
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ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Tingting Zhao,Min Zhang,Qi Peng,Xucong Lin,Zenghong Xie
Herein, a facile protocol of simple DNA adsorption on UV-initiated polymerization supports was proposed for effectively fabricating aptamer-based affinity monolithic column. Hydrophilic cationic monolith with an excellent mechanical stability was achieved within 7 min and then massive aptamers were directly bound by DNA charge-dependent adsorption. Strong cationic quaternary ammonium-based monomer was employed to provide effective and stable positive charge surface for aptamer immobilization in a wide range of pH. An ultra-high aptamer coverage density of 6813 pmol/μL was achieved to gain a highly specific online recognition performance. Limitations such as low aptamer capacity, tedious modification and time-consuming reactions in the traditional biological or covalent modification strategies were avoided. By using ochratoxin A (OTA) as the given analyte, the selective recognition and high recoveries were successfully achieved, and little cross-reactivity towards OTB analogue was only 0.5% even if the content of OTB got up to 125 folds of OTA. Applied to sample analysis, the satisfactory discriminations of trace OTA were obtained at 93.9 ± 1.9% – 96.5 ± 1.7%（n = 3）in beer, wheat and chicken liver samples. It might light a cost-effective access to efficiently preparing high-performance affinity monoliths towards the selective in-tube microextraction of OTA.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Wanqiao Bai,Wenyu He,Huan Wang,Sijia Li,Xia Yang,Huiping Sun,Yan Li
5-Hydroxymethylcytosine (5-hmC), as the sixth base in the mammalian genome, is increasingly regarded as an epigenetic marker with crucial biological functions. Herein, a highly sensitive electrogenerated chemiluminescence (ECL) sensing platform for double-stranded DNA containing 5-hmC (5-hmC-dsDNA) in genome was fabricated, which based on polyadenine-gold interaction combining with signal amplification of hybridization chain reaction (HCR). This biosensing strategy could be executed to detect 5-hmC in the native double-stranded DNA, irrespective of the specific sequences of DNA. Firstly, DNA methyltransferase (M.HhaI) was introduced to chemically modify the hydroxyl of 5-hmC in double-stranded DNA, which converted 5-hmC-dsDNA to amino-derived 5-hmC-dsDNA. Subsequently, DNA sequence containing polyadenine (polyA) was used as an anchor probe to incorporate the derivatized 5-hmC-dsDNA by amide reaction to form compound DNA and then completely assembled onto the gold electrode surface through preferential polyA-gold interaction. Subsequently, two designed DNA hairpin probes which can alternately hybridize with each other were linked to the signal amplification part of multifunctional polyA probes, forming ultra-long nicked double helix structures. Finally, ECL indicator of Ru(phen) 3 2+ molecules were intercalated into the groove of the nicked DNA double helices, resulting greatly increased ECL intensity. Therefore, the quantitatively analysis of 5-hmC-dsDNA could be obtained relied on the relationship between the ECL intensity and 5-hmC-dsDNA content. The detection limit of this method for 5-hmC-dsDNA was 0.0001 % (S/N = 3). Moreover, the biosensing platform was successfully applied to detect 5-hmC-dsDNA in mice tissue samples with satisfactory results, manifesting its great potential application in clinical analysis.
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