Analytical Methods
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2016
·Vol. 8
·期 48
·8461-8465
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IF 1.915
Hongyan Wang,Jine Wang,Lijun Xu,Yajie Zhang,Yang Chen,Hongxia Chen,Renjun Pei
Aptamers are single-stranded oligonucleotides which are selected and are capable of binding certain targets tightly and specifically. Systematic Evolution of Ligands by Exponential Enrichment (SELEX) has been used to select aptamers for various targets such as proteins, metal ions, small molecules and so on. For the first time, we have used a modified SELEX technology to obtain DNA aptamers for a fluorogenic dye, thioflavin T (ThT). DNA aptamers with low micromolar dissociation constants were achieved after only 8 rounds of selection and the final minimized 33-mer aptamer showed 191-fold fluorescence enhancement in a saturated concentration. The truncated DNA aptamer showed excellent selectivity and specificity for ThT.
研究领域:
分子生物学
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生物化学
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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
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分析化学
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PubMed
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CHINESE CHEMICAL LETTERS
·
2021
·
IF 6.779
Yuan Liu,Gaojian Yang,Taotao Li,Yan Deng,Zhu Chen,Nongyue He
An improved ssDNA library immobilized systematic evolution of ligands by enrichment (SELEX) was applied to select aptamers against carbaryl. After nine selection rounds, a highly enriched ssDNA pool was obtained. The Apta3 was demonstrated as the optimal aptamer. In order to facilitate the modification of aptamer, the Apta3 was further truncated with the dissociation constant ( K d ) of 0.364 ± 0.055 μmol/L and a fluorescent aptasensor was developed. The linear range for carbaryl was from 100 nmol/L to 1500 nmol/L, with the limit of detection was as low as 15.23 nmol/L. Besides, the biosensor was validated for the carbaryl spiked real samples, and the recoveries were between 97.7% and 107.3%.
研究领域:
生物化学
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分析化学
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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
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分析化学
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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
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分子生物学
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食品安全
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JOURNAL OF HAZARDOUS MATERIALS
·
2025
·
IF 11.3
Lingjun Geng,Jingcheng Huang,Haowei Dong,Haifang Wang,Rui Xu,Zhiping Yu,Ibrahim A. Darwish,Yemin Guo,Xia Sun
Organophosphorus pesticides (OPs) have attracted attention due to their widespread application in agriculture and public health sector. Among them, azamethiphos (AZA) may pose risks to non-target organisms and human health through food chain accumulation. Therefore, establishing a highly sensitive and specific detection method of AZA is of great significance for ensuring food safety and ecological protection. In this study, based on graphene oxide-SELEX technology, an aptamer (Apt) with specific recognition for AZA (K d =26.27±1.27 nM) was screened for the first time. This aptamer was subsequently integrated with molecularly imprinted polymers to construct a dual-recognition electrochemical sensor, leveraging the complementary advantages of both recognition elements. This dual-recognition strategy enabled the synergistic enhancement of specific recognition ability, effectively reducing interference from non-target substances and improving sensor selectivity and accuracy. Additionally, a doping strategy was adopted to modify the electrode surface with AuNPs@SnS 2 @ZnCo-MOF nanocomposites, improving electron transfer efficiency and providing abundant active sites, thereby significantly enhancing the electrochemical signal response. The sensor exhibited a wide detection range (1.00×10 −2 -1.00×10 4 ng/mL) and a low limit of detection (3.33×10 −3 ng/mL), while also demonstrating excellent stability and specificity. In summary, this study developed a highly efficient, sensitive and selective electrochemical sensor, providing a novel strategy for the rapid detection of AZA and other organophosphorus pesticides, with broad application prospects in food safety and environmental monitoring.
研究领域:
食品安全检测
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环境监测
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生物传感技术
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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
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分析化学
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药理学
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ANALYTICA CHIMICA ACTA
·
2017
·
IF 4.95
Nuo Duan,Wenhui Gong,Shijia Wu,Zhouping Wang
An improved SELEX technique was developed for selecting aptamers against ractopamine (RAC) by immobilizing ssDNA library on the magnetic beads. After sixteen selection rounds, a highly enriched ssDNA pool was sequenced and nine families were grouped according to their homology and secondary structures analysis. One representative aptamer candidate from each family was picked out for binding affinity identification by graphene oxide (GO) adsorption platform. The aptamer RAC-6 was demonstrated as the optimal aptamer with high specificity and dissociation constant (K d ) value of 54.22 ± 8.02 nM. To prove the potential application of aptamer RAC-6 in the quantitative determination of RAC, a fluorescent bioassay with aptamer RAC-6 was developed. The linear range for RAC was from 0.10 ng/mL to 100 ng/mL and the limit of detection was as low as 0.04 ng/mL. Furthermore, the method was validated for the analysis of RAC spiked real samples, and the recoveries were between 82.57% and 104.65%.
研究领域:
分子生物学
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分析化学
·
生物技术
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PubMed
高影响因子
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