ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Yuxia Yang,Yue Tang,Chunxiao Wang,Bangyan Liu,Yuangen Wu
Pyrethroid pesticides residues will not only pollute the environment, but also cause high toxicity to the human body. It is significant to establish an efficient and accurate method for pyrethroid detection in food. Considering that the common biomolecules like antibody is complicated and easy to inactivate, it is urgent to find a new type of biomolecule to specifically recognize pyrethroid pesticides. This study proposed the Capture-SELEX strategy to firstly select λ-cyhalothrin aptamer by immobilizing random ssDNA library. High-throughput sequencing was performed on the enriched ssDNA library through multiple Capture-SELEX rounds. Comprehensively inspecting structural similarity and homology , six sequences were chosen from five families for further analysis. The results showed that the aptamer (named LCT-1) could specifically recognize λ-cyhalothrin with the strongest affinity ( K d = 50.64 ± 4.33 nmol L −1 ). Molecular docking results revealed that the binding sites between λ-cyhalothrin and LCT-1 aptamer are mainly related to the bases A-5, C-6, C-28, A-29, C-30, G-31 and G-32. The LCT-1 aptamer was truncated to a shorter sequence (named as LCT-1-39) by removing other irrelevant bases, and its K d value was determined as (10.27 ± 1.33) nmol·L −1 by Microscale Thermophoresis (MST). Both LCT-1 and LCT-1-39 aptamers were employed as recognition molecules to establish the colorimetric aptasensors for λ-cyhalothrin detection, which displayed good repeatability and reproducibility. The detection limit of the aptasensors were individually calculated as 0.0197 μg ml −1 and 0.0186 μg ml −1 , and their recovery rate of λ-cyhalothrin in pear and cucumber samples was in the range of 82.93–95.50%. This article provides a promising application for the detection of λ-cyhalothrin.
研究领域:
食品安全检测
·
分子生物学
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
·
IF 4.4
Xuanping Tan,Yuhui Zhang,Huaping Mao,Jidong Yang
In this work, specific aptamers with affinity for S-propranolol were screened by SELEX technology based on the graphene oxide (GO) adsorption platform, and a GO-FAM labeled aptamer-propranolol fluorescent optical switch system was constructed for the recognition of chiral propranolol. It was found that the fluorescence quenching of FAM labeled aptamer could be caused by the adsorption of GO. However, when S-propranolol was introduced, S-propranolol could pull out the aptamer adsorbed by GO, and the fluorescence of the system could be restored. But, R-propranolol could not be realized. Therefore, a simple and sensitive fluorescent optical switch system was established to identify chiral propranolol and perform highly sensitive detection of S-propranolol.
研究领域:
生物化学
·
分析化学
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DOI
PubMed
高影响因子
Analytical Methods
·
2021
·Vol. 13
·期 27
·3110-3117
·
IF 2.896
Mingyan Song,Chen Lyu,Nuo Duan,Shijia Wu,Imran Mahmood Khan,Zhouping Wang
The abuse of ribavirin, an antiviral drug, in poultry breeding can cause quality degradation and drug resistance. So it is of great importance to establish a simple and effective method for detecting ribavirin in foods. In this work, aptamers that could especially bind to ribavirin with high affinity were obtained by the Capture-SELEX method. After 15 rounds of enrichment, ssDNA library pool was enriched and then analyzed by high-throughput sequencing. The seven most enriched sequences were selected as candidate aptamers for affinity and specificity characterization. Among the candidate aptamers, APT-1 was proved to be the optimal aptamer. The dissociation constant (Kd) values of APT-1 obtained by the two methods of colorimetric and fluorescence were 34.34 ± 6.038 nmol L−1, 61.19 ± 21.48 nmol L−1, respectively. To study the binding mechanism of the selected aptamer, molecular docking was conducted and results indicated that hydrogen bonds were formed at binding sites located at G37, T38, A40, T53 and A54. Furthermore, to confirm the practicability of the selected aptamer, a fluorescence assay was designed, showing the liner range within 1.0–50 ng mL−1 and the low detection limit of 0.67 ng mL−1. Besides, the aptamer was applied for the detection of ribavirin in chicken samples and the recoveries ranged from 87.26% to 105.57%, which showed great application potential in food safety.
研究领域:
分析化学
·
食品科学
·
分子生物学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2026
·
IF 6.1
Xue Wang,Zhaoyang Zhou,Shuang Jiang,Qi Sun,Xianlu Lei,Yong Xie,Tao Le
Norfloxacin (NOR) has been widely applied in animal husbandry and aquaculture because of its potent antibacterial activity, broad-spectrum efficacy and cost-efficient. Nevertheless, its inappropriate use may result in teratogenic and carcinogenic risks to humans, making the accurate and rapid detection of NOR residues in food critically important. In this study, a strong affinity (Kd = 84 nM) and high selectivity anti-NOR aptamer was obtained by screening with Graphene Oxide Systematic Evolution of Ligands by Exponential Enrichment (GO-SELEX), and a colorimetric aptasensor was fabricated. The limit of detection of the aptasensor was 1.04 ng/mL, and the linear range was 1.56-100 ng/mL. Average recoveries of the aptasensor in shrimp and fish sample analyses ranged from 89.33% to 103.24%, accompanied by coefficients of variation from 3.63% to 9.43%, indicating good analytical precision and accuracy. The aptasensor constructed in this study showed high consistency with the High-Performance Liquid Chromatography (HPLC) for NOR detection, indicating that it is an efficient and promising sustainable strategy for monitoring NOR.
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PubMed
高影响因子
MOLECULES
·
2018
·Vol. 23
·期 9
·2337
·
IF 3.098
Xixia Liu,Qi Lu,Sirui Chen,Fang Wang,Jianjun Hou,Zhenlin Xu,Chen Meng,Tianyuan Hu,Yaoyao Hou
We describe a multiple combined strategy to discover novel aptamers specific for clenbuterol (CBL). An immobilized ssDNA library was used for the selection of specific aptamers using the systematic evolution of ligands by exponential enrichment (SELEX). Progress was monitored using real-time quantitative PCR (Q-PCR), and the enriched library was sequenced by high-throughput sequencing. Candidate aptamers were picked and preliminarily identified using a gold nanoparticles (AuNPs) biosensor. Bioactive aptamers were characterized for affinity, circular dichroism (CD), specificity and sensitivity. The Q-PCR amplification curve increased and the retention rate was about 1% at the eighth round. Use of the AuNPs biosensor and CD analyses determined that six aptamers had binding activity. Affinity analysis showed that aptamer 47 had the highest affinity (Kd = 42.17 ± 8.98 nM) with no cross reactivity to CBL analogs. Indirect competitive enzyme linked aptamer assay (IC-ELAA) based on a 5′-biotin aptamer 47 indicated the limit of detection (LOD) was 0.18 ± 0.02 ng/L (n = 3), and it was used to detect pork samples with a mean recovery of 83.33–97.03%. This is the first report of a universal strategy including library fixation, Q-PCR monitoring, high-throughput sequencing, and AuNPs biosensor identification to select aptamers specific for small molecules.Keywords:clenbuterol;systematic evolution of ligands by exponential enrichment;real-time quantitative PCR;high-throughput sequencing technology;aptamers;gold nanoparticles biosensor
研究领域:
生物技术
·
分析化学
·
纳米技术
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分子生物学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Tianyu Huang,Xin Chen,Jinri Chen,Yuting Zhang,Xiaoli Wang,Zhimeng Wu,Nandi Zhou
Azithromycin can effectively inhibit bacterial protein synthesis and reduce biofilm formation and quorum sensing, and thus has been preferentially used in the treatment of various infections. However, azithromycin is also among the most highly concentrated antibiotics present in wastewater due to the inappropriate use and treatment. In order to sensitively and rapidly monitor the azithromycin residues, aptamers specific to azithromycin were selected through 12 rounds of screening by using magnetic beads-SELEX (MB-SELEX). The screened aptamers were assessed for their affinity and specificity using fluorescent assay. Apt3 emerged as the optimal candidate aptamer with the dissociation constant (K d ) of 235.07 nM. After secondary structure analysis and molecular docking, Apt3 was systematically truncated and subjected to single-base mutations. The optimal aptamer Apt3–27 T was identified, exhibiting the K d value of 215.84 nM and the improved specificity compared to the original aptamer. Then Apt3–27 T was utilized to construct a ratiometric fluorescent aptasensor, which exhibited the linear dynamic range of 25–400 nM and low detection limit of 9.78 nM. More importantly, the ratiometric fluorescence mechanism further improved the specificity of the biosensor, which finally can discriminate azithromycin from structurally similar erythromycin. Evaluation experiments using actual samples spiked with azithromycin showed the recovery ranging from 97.1% to 106% and the relative standard deviation ranging from 4.29% to 8.61%. The screened aptamer and constructed aptasensor offer the advantages of high affinity, high effectiveness and ease of use in actual sample detection, thereby displaying promising applications in the monitoring of azithromycin residues.
研究领域:
分析化学
·
环境科学
·
生物传感器
·
药物分析
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DOI
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Xiaoling Zheng,Shuang Jiang,Yueyang Ren,Sixian Wang,Yong Xie,Tao Le
Accurate and rapid detection of nitrofurantoin metabolite 1-aminohydantoin (AHD), as one of the tissue-bound toxic metabolites in foods, is crucial. In this study, a fluorescent aptasensor was constructed for the detection of AHD, using anti-AHD aptamers firstly obtained by high-efficiency magnetic crosslinking precipitation (MCP)-SELEX. The AHD-specific aptamers were optimized and identified for their binding affinity and specificity using fluorescence-based assays and molecular dynamics simulations. The truncated aptamer AHD50-1, which exhibited highest affinity ( K d = 86 nM) and an excellent specificity, was applied to the construction of fluorescent aptasensor. Under optimized conditions, the aptasensor showed a low limit of detection (0.2 ng/mL) and a wide dynamic linear range (0.25–150 ng/mL). For the five types of samples (pork, beef, shrimp, fish and honey), the recoveries ranged from 89.4% to 110.04%, with coefficients of variation ranging from 1.43% to 14.46%. These results obtained by the proposed aptasensor corroborate very well with the method of HPLC for detection of AHD, indicating that the developed aptasensor provides an efficient and promising sustainable strategy for monitoring AHD in animal derived food.
研究领域:
分析化学
·
分子生物学
·
食品安全
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Wenwei Wu,Lixia Qian,Jie Shen,Hongmei Li,Liang-cheng Li
Cancer remains a leading cause of mortality worldwide, with China bearing a particularly high burden. Early detection through reliable biomarkers is essential for improving patient outcomes. This study reports the development and validation of a highly specific aptamer-based biosensor for detecting the complement protein C1QC, a promising cancer biomarker. Using a multi-step selection process involving Magnetic beads-SELEX, qPCR, flow cytometry, high-throughput sequencing, surface plasmon resonance (SPR), pull-down, and western blotting, we identified Apt 13, a slow-dissociation aptamer with high specificity and affinity for C1QC (K D = 48.7 nM). SPR confirmed the formation of a sandwich complex between Apt 13, C1QC, and Anti_C1QC, confirming their distinct binding sites. We developed two biosensor platforms based on this aptamer: the enzyme-linked antibody-aptamer sandwich (ELAAS) and chemiluminescent antibody-aptamer sandwich (CLAAS) biosensors. The ELAAS biosensor, designed for visual colorimetric detection, demonstrated a limit of detection (LOD) of 98 ng/mL. In contrast, the CLAAS system, utilizing acridinium ester-modified antibodies, achieved an enhanced LOD of 0.375 ng/mL. Receiver Operating Characteristic (ROC) analysis on serum samples from 75 hepatocellular carcinoma (HCC) patients and 20 healthy controls yielded an AUC of 0.942, with 86.67 % sensitivity and 95.00 % specificity, demonstrating CLAAS’s high diagnostic accuracy in differentiating HCC from controls. Further testing in HCC patient serum samples revealed significantly elevated C1QC levels compared to healthy controls, underscoring C1QC’s potential as a HCC cancer biomarker. Apt 13 presents a promising tool for the development of innovative, sensitive, and cost-effective diagnostic technologies for C1QC detection and may offer future potential for targeted therapies.
研究领域:
癌症研究
·
分子诊断
·
生物传感器
·
适配体技术
·
蛋白质组学
·
临床诊断
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DOI
高影响因子
Food Analytical Methods
·
2022
·Vol. 16
·期 1
·215-224
·
IF 3.498
Sha Junyi,Lin Hong,Zhang Ziye,Sui Jianxin
Aptamer is widely used in the detection field as a new type of probe due to their advantages of easy synthesis and modification. Due to the traditional aptamer screening method, the aptamers that are screened by SELEX technology generally have redundant sequences, which affect the binding affinity and detection sensitivity. In this study, a structural transformation strategy for rationally shortening was established by analyzing the secondary structure, finding the sequence of the binding site by base mutation, and double cloning the binding sites. We obtained a 27-mer new aptamer SEQ.A52 with the dissociation constants (Kd) of 10.74 nM, which is 21 times higher than the original aptamer. When used to detect enrofloxacin (ENR) with the fluorescent method, the sensitivity could reach 50 to approximately 1000 pM, and the limit of detections (LOD) was only 41.35 pM, which was 88 times higher than the original aptamer. All the results implied that the strategy could be used for structural modification of some other aptamers to increase affinity and sensitivity.
研究领域:
生物技术
·
分析化学
·
分子生物学
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DOI
高影响因子
Signal Transduction and Targeted Therapy
·
2024
·Vol. 9
·期 1
·1-13
·
IF 40.8
Yang Minghui,Li Chunhui,Ye Guoguo,Shen Chenguang,Shi Huiping,Zhong Liping,Tian Yuxin,Zhao Mengyuan,Wu Pengfei,Hussain Abid,Zhang Tian,Yang Haiyin,Yang Jun,Weng Yuhua,Liu Xinyue,Wang Zhimin,Gan Lu,Zhang Qianyu,Liu Yingxia,Yang Ge,Huang Yuanyu,Zhao Yongxiang
Emerging and recurrent infectious diseases caused by human coronaviruses (HCoVs) continue to pose a significant threat to global public health security. In light of this ongoing threat, the development of a broad-spectrum drug to combat HCoVs is an urgently priority. Herein, we report a series of anti-pan-coronavirus ssDNA aptamers screened using Systematic Evolution of Ligands by Exponential Enrichment (SELEX). These aptamers have nanomolar affinity with the nucleocapsid protein (NP) of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and also show excellent binding efficiency to the N proteins of both SARS, MERS, HCoV-OC43 and -NL63 with affinity K D values of 1.31 to 135.36 nM. Such aptamer-based therapeutics exhibited potent antiviral activity against both the authentic SARS-CoV-2 prototype strain and the Omicron variant (BA.5) with EC 50 values at 2.00 nM and 41.08 nM, respectively. The protein docking analysis also evidenced that these aptamers exhibit strong affinities for N proteins of pan-coronavirus and other HCoVs (−229E and -HKU1). In conclusion, we have identified six aptamers with a high pan-coronavirus antiviral activity, which could potentially serve as an effective strategy for preventing infections by unknown coronaviruses and addressing the ongoing global health threat.
研究领域:
病毒学
·
分子生物学
·
药物开发
·
传染病
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DOI
PubMed
高影响因子
TALANTA
·
2022
·
IF 6.556
Mingqian Song,Changxin Li,Shijia Wu,Nuo Duan
Chlorpromazine is a phenothiazine representative drug that can be used to anesthetize and calm animals. However, chlorpromazine excess may lead to residual persistence in edible tissues, which is potentially harmful to human health and animal production. In this work, high affinity and specificity aptamers against chlorpromazine were screened out based on Capture-SELEX. After ten rounds of screening, the candidate aptamers were obtained. The optimal aptamer of CHL-3 was obtained by isothermal titration calorimetry (ITC) and SYBR Green I (SGI) fluorescence-competition methods. The Kd value of CHL-3 was 69.8 ± 9.81 nM. Subsequently, the Uio-66-NH 2 material was prepared, filled with rhodamine 6G (Rho 6G) dye into the pore, and sealed with CHL-3, and fluorescence probes were obtained. The ratiometric fluorescence detection method was established to detect the concentration of chlorpromazine. A linear relationship was obtained in a range of 1–100 nM, with a lower detection limit of 0.67 nM. Meanwhile, a good recovery was shown in spiked food samples, such as eggs and milk. These results indicate that the constructed ratiometric fluorescence strategy can be successfully applied in food detection.
研究领域:
分析化学
·
生物传感器
·
食品安全
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 19
·4855-4863
·
IF 4.157
Ren Le,Qi Shuo,Khan Imran Mahmood,Wu Shijia,Duan Nuo,Wang Zhouping
Acyclic guanosine analogues, a class of widely used antiviral drugs, can cause chronic toxicity and virus resistance. Therefore, it is essential to establish rapid and accurate methods to detect acyclic guanosine analogues. In this study, five acyclic guanosine analogues (acyclovir, famciclovir, ganciclovir, penciclovir, and valaciclovir) were used as positive targets to obtain broad-spectrum aptamers through Capture-SELEX technology. Real-time quantitative PCR (Q-PCR) was used to monitor the aptamer SELEX process. After the sixteen rounds of selection against mixed targets, sequences were obtained by high-throughput sequencing (HTS). Furthermore, a broad-spectrum aptamer, named CIV6, was found as the higher performance aptamer that was suitable for five acyclic guanosine analogues by graphene oxide (GO) polarization and fluorescence assay. Finally, the aptamer CIV6 was used to construct GO fluorescence assay to detect five acyclic guanosine analogues. The limits of detection (LOD) of acyclovir, famciclovir, ganciclovir, penciclovir, and valaciclovir were 0.48 ng·mL −1 , 0.53 ng·mL −1 , 0.50 ng·mL −1 , 0.56 ng·mL −1 , and 0.38 ng·mL −1 , respectively. Graphical abstract
研究领域:
分子生物学
·
分析化学
·
药理学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）