MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Yongshu Li,Yani Li,Qin Wu,Jianjun Hou,Xixia Liu
As a promising recognition molecule, aptamers are widely used in the field of small molecule detection. However, few reports are available on the key factors governing successful aptamer selection. To improve the success rate, the melamine-specific aptamer served as a model to investigate critical factors in library-immobilization-based SELEX methods (GO-SELEX and Beads-SELEX). The results indicated that non-ionic surfactants significantly enhanced the desorption of ssDNA from the graphene oxide (GO) surface in Mode one of the mimic GO-SELEX (GO-facilitated library immobilization and target-induced aptamer release). In contrast, in Mode two of the mimic GO-SELEX, where GO was used solely for non-target-binding aptamer removal, and significant non-specific binding between ssDNA and GO compromised the enrichment efficiency of target-specific sequences, necessitating the use of high target concentrations during SELEX. Similarly, in Beads-SELEX, high target concentrations were required to release specific ssDNA, which also led to a small amount of non-specific ssDNA co-release during the procedures. Other SELEX conditions were also compared and constructive suggestions were provided for facilitating the successful aptamer selection. In summary, this study provided SELEX condition optimization guidance for the selection of aptamers specific to small molecules, when different library-immobilized SELEX methods were performed.
研究领域:
生物化学
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分子生物学
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分析化学
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生物技术
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高影响因子
Food Analytical Methods
·
2022
·Vol. 15
·期 6
·1535-1544
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IF 3.498
Li Linfang,Zhou Jinru,Wang Kuiwu,Chen Xiaojing,Fu Linglin,Wang Yanbo
Tropomyosin (TM), known as the major allergen in shellfish, has gained increasing concerns in industrialized countries, resulting in an urgent need for an accurate, simple, and sensitive method for TM detection in food products. Nucleic acid aptamers, short single-stranded oligonucleotides chains, can bind to various targets with a high affinity and specificity and hence have the potential to be used for the accurate detection of targets. Here, we proposed capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX) for rapid selection and identification of specific aptamers against shellfish TM. A selected aptamer IX exhibited high specificity and high binding affinity against TM with K d determined (0.95 nM). In addition, the aptamer IX has great potential to be employed in the establishment of various aptasensors for TM detection in shellfish products. Furthermore, the CE-SELEX method may provide a universal and convenient method for the aptamer selection against more targets.
研究领域:
食品科学
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分子生物学
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高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2017
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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
高影响因子
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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DOI
PubMed
高影响因子
SUPRAMOLECULAR CHEMISTRY
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2025
·
IF 2.6
Peng Chen,Lancheng Wang,Dandan Xue,Mingjie Xu,Chi Hu
Split aptamers (SPAs) display unique functions to bind one another in the presence of a specific small-molecule or protein target, thus recognised as compelling tools in diagnostics and therapy. However, only a limited number of SPAs have been proven successful so far due to the largely compromised affinity, specificity and stability arising from cleaving a long parent aptamer. To this end, we propose a supramolecular anchoring approach to improve SPA function after the Systematic Evolution of Ligands by Exponential Enrichment process. With the macrocyclic host cucurbituril and its guest molecule tethered to separate strands of an SPA for the fluorescence dye Thioflavin T (TT), the anchoring module was constructed by host-guest complexation once TT was bound to the SPA and brought the discrete aptamer units into proximity. With the supramolecular anchor to facilitate the aptamer-target complex formation and stabilise the assembled structure via dynamic crosslinking, SPA function for bio-imaging and sensing purposes was largely improved as witnessed by the increased fluorescence readout of TT in the aptamer-bound form, even when the nucleic acid sequences were shortened below the duplex-formation level. The obtained fluorescence response tracked well with the anchoring strength (none, weak or strong), thereby validating the supramolecular entity as a sophisticated and versatile strategy to rehash the otherwise incompetent SPAs.
研究领域:
生物化学
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分子生物学
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纳米技术
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生物医学工程
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分析化学
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高影响因子
TALANTA
·
2018
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IF 4.244
Junqing He,Junyan Wang,Nan Zhang,Luyao Shen,Linlin Wang,Xiao Xiao,Yan Wang,Tao Bing,Xiangjun Liu,Songqing Li,Dihua Shangguan
Ovarian cancer is regarded as the most lethal gynecologic malignancy with poor prognosis and high mortality rate. Drug-resistance was thought to be the main obstacle to improving overall survival rate of ovarian cancer. New ligands for drug-resistant ovarian cancer cells have potential for the development of diagnosis and therapy of ovarian cancer. In present work, we reported two aptamers , HF3–58 and HA5–68 generated by cell-SELEX, against a paclitaxel-resistant ovarian cancer cell line (A2780T). Both two aptamers exhibited high selectivity and strong affinity to target cells with low nanomolar dissociation constants . The binding of aptamers to target cells was independent of divalent ions, and was tolerant of incubation temperature and nucleases in serum. Molecular targets of the two aptamers were preliminarily demonstrated to be two different glycoproteins on cell surface of A2780T cells. Owing to the structure stability and high resistance to nuclease, these two aptamers had good performance in the detection of drug-resistant ovarian cancer cells in human serum.
研究领域:
癌症生物学
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分子生物学
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药学
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PubMed
高影响因子
TALANTA
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2025
·
IF 6.1
Lian Xia,Jia Zheng,Jian Su,Yue Tang,Zhen Wu,Xiaojuan Niu,Yuangen Wu
Amygdalin, a cyanogenic glycoside abundant in apricot kernels, poses a food safety risk, requiring rapid detection that conventional chromatographic methods cannot fulfill for on-site application. Thus, this study aims to construct a novel colorimetric sensing platform and improve detection performance through a multivalent aptamer strategy. This pioneering work introduces a novel bivalent aptamer strategy for ultra-sensitive on-site amygdalin detection. The aptamers specific to amygdalin were screened using a five-segment single-stranded DNA library and Capture-SELEX, with truncated variants optimized to obtain a high-affinity core sequence. A key innovation involves the first design and synthesis of a bivalent aptamer that exploits synergistic binding to enhance target affinity. The bivalent aptamer (Bapt 4-4) exhibited doubled affinity for amygdalin compared to its monovalent truncated counterpart (Amyg 4–26), achieving a dissociation constant ( K d ) of (2.19 ± 1.03) nM. By integrating the bivalent aptamer with the laccase-like activity of silver oxide nanoparticles (Ag 2 O NPs), an aptasensor with high sensitivity and specificity was developed, yielding a detection limit of 0.008 μg mL −1 , which is lower than that of the monovalent aptamer-based sensor (0.0137 μg mL −1 ). This multivalent aptamer strategy provides an effective approach for highly sensitive amygdalin detection. Furthermore, the constructed bivalent aptasensor provides a robust technical tool for the on-site monitoring of plant-derived cyanogenic glycoside toxins, with broad application prospects in the domains of food safety and traditional Chinese medicine safety.
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高影响因子
ANALYTICA CHIMICA ACTA
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2017
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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
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生物技术
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PubMed
高影响因子
ANNALS OF THE RHEUMATIC DISEASES
·
2024
·
IF 20.3
Fang Qiu,Duoli Xie,Hongzhen Chen,Zhuqian Wang,Jie Huang,Chunhao Cao,Yiying Liang,Xu Yang,Dong-Yi He,Xuekun Fu,Aiping Lu,Chao Liang
Objectives Rheumatoid arthritis (RA) is an autoimmune disease characterised by aggressive fibroblast-like synoviocytes (FLSs). Very few RA patients-derived FLSs (RA-FLSs)-specific surface signatures have been identified, and there is currently no approved targeted therapy for RA-FLSs. This study aimed to screen therapeutic aptamers with cell-targeting and cytotoxic properties against RA-FLSs and to uncover the molecular targets and mechanism of action of the screened aptamers. Methods We developed a cell-specific and cytotoxic systematic evolution of ligands by exponential enrichment (CSCT-SELEX) method to screen the therapeutic aptamers without prior knowledge of the surface signatures of RA-FLSs. The molecular targets and mechanisms of action of the screened aptamers were determined by pull-down assays and RNA sequencing. The therapeutic efficacy of the screened aptamers was examined in arthritic mouse models. Results We obtained an aptamer SAPT8 that selectively recognised and killed RA-FLSs. The molecular target of SAPT8 was nucleolin (NCL), a shuttling protein overexpressed on the surface and involved in the tumor-like transformation of RA-FLSs. Mechanistically, SAPT8 interacted with the surface NCL and was internalised to achieve lysosomal degradation of NCL, leading to the upregulation of proapoptotic p53 and downregulation of antiapoptotic B-cell lymphoma 2 (Bcl-2) in RA-FLSs. When administrated systemically to arthritic mice, SAPT8 accumulated in the inflamed FLSs of joints. SAPT8 monotherapy or its combination with tumour necrosis factor (TNF)-targeted biologics was shown to relieve arthritis in mouse models. Conclusions CSCT-SELEX could be a promising strategy for developing cell-targeting and cytotoxic aptamers. SAPT8 aptamer selectively ablates RA-FLSs via modulating NCL-p53/Bcl-2 signalling, representing a potential alternative or complementary therapy for RA.
研究领域:
免疫学
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分子生物学
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治疗学
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自身免疫疾病
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高影响因子
FOOD CHEMISTRY
·
2023
·
IF 8.8
Ying Li,Boyu Jia,Pengyue Song,Nan Long,Linchun Shi,Peng Li,Jiabo Wang,Lidong Zhou,Weijun Kong
As nucleic acid-based affinity elements, aptamers have attracted significant attention for a wide range of analytical applications. Although several aflatoxin B 1 (AFB 1 ) aptamers have been identified, they are unsuitable for overcoming the unavoidable cross-reactions from interferents in complex food matrices due to their poor binding affinities and specificities. Herein, a novel precision-systematic evolution of ligands by exponential enrichment (P-SELEX) strategy through introducing the counter (matrix without target AFB 1 ) and positive (with AFB 1 ) screening steps was implemented to accurately identify AFB 1 aptamers. A DNA aptamer A-42-2 at a 24-nt length was selected finally, which possessed nanomolar-level affinity of 5.55 nM, high specificity to other interferents, and strong anti-cross-reactivity ability for matrix components. Then, an A-42-2 aptamer-based ultra-sensitive colorimetric and fluorescent dual-readout aptasensor was fabricated for AFB 1 detection in three kinds of complex food samples rich in starch without cross-reactions. The aptasensor displayed outstanding detection capacity with a wide liner range of 0.25–30 nM (1.95–234.4 μg/kg), while the detection limit for colorimetric measurement as low as 0.22 nM (1.72 μg/kg) and 0.048 nM (0.20 μg/kg) for fluorescent determination. P-SELEX is ideal for screening and applying aptamers in complex food matrices, creating more opportunities for the efficient and cost-effective development of high-quality aptamers and aptasensors for other targets.
研究领域:
分析化学
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食品科学
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生物技术
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分子生物学
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PubMed
高影响因子
RSC Advances
·
2019
·Vol. 9
·期 11
·6328-6334
·
IF 3.049
Qiong-Lin Wang,Hui-Fang Cui,Jiang-Feng Du,Qi-Yan Lv,Xiaojie Song
DNA aptamers against carcinoembryonic antigen (CEA) have been identified through the systematic evolution of ligands by exponential enrichment (SELEX) technique, but their affinity needs to be improved. In this study, an in silico approach was firstly used to screen the mutation sequences of a reported DNA aptamer (the parent aptamer, denoted as P) against CEA. The affinities of several high-score DNA mutants were determined by the biolayer interferometry technique. Finally, the newly obtained aptamers were verified in an aptasensor application. For the in silico approach, Mfold and RNA Composer were combined to generate the 3D RNA structures of the DNA mutants. The RNA structures were then modified to 3D DNA structures with the Write program. The docking model and binding ability of the 3D DNA structures with CEA were simulated and predicted with the ZDOCK program. Two mutation sequences (P-ATG and GAC-P) exhibited significantly higher ZDOCK scores than P. The dissociation constant of P-ATG and GAC-P to CEA was determined to be 4.62 and 3.93 nM respectively, obviously superior to that of P (6.95 nM). The detection limit of the P-ATG and GAC-P based aptasensors was 1.5 and 1.2 ng mL−1, respectively, markedly better than that based on P (3.4 ng mL−1). The consistency between the in silico and the experimental results indicates that the developed in silico post-SELEX screening approach is feasible for improving DNA aptamers. The P-ATG and GAC-P aptamers found in this study could be used for future CEA aptasensor design and fabrication, promisingly applicable for highly sensitive CEA detection and early cancer diagnosis.
研究领域:
生物信息学
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分子生物学
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癌症诊断
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纳米技术
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2022
·
IF 5.895
Mengyue Liu,Lingjun Geng,Fengjuan Zhang,Shouyi Dou,Falan Li,Zhanli Liu,Yemin Guo,Xia Sun
In this work, an aptamer against Escherichia coli is isolated via non-SELEX, which executes efficient selection by employing repetitive cycles of centrifugation-based partitioning, and the binding site of the aptamer on E. coli cell surfaces is inferred to be a membrane protein. Moreover, truncated sequence 2–17–2 with a higher affinity (Kd = 101.76 nM) is employed for highly sensitive colorimetric detection of bacteria based on the dual signal amplification strategy. When targets exist, the release of DNA 1 from the polymer activates a hybridization chain reaction (HCR) between DNA 1 and DNA 2, thereby inducing the aggregation of probe 1. Subsequently, DNA 3 dissociated from probe 1 as a linker DNA further assembles probe 2/3. In this system, two types of DNA@gold nanoparticles (AuNPs) coexist and successively aggregate AuNPs based on divergent triggering mechanisms. Under optimal conditions, the dual signal amplification strategy presents excellent sensitivity (10 CFU mL–1) and specificity, as well as the realization of real sample analysis.
研究领域:
微生物学
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纳米技术
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分析化学
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生物技术
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DOI
PubMed
高影响因子
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