ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Lian Xia,Yuxia Yang,Hongqin Yang,Yue Tang,Jianli Zhou,Yuangen Wu
There is still a great challenge of on-site and rapid detection for ethyl carbamate (EC) in liquor. Herein, the aptamer (EC1) against EC was firstly screened by Capture-SELEX strategy, and its affinity and binding sites for EC were further investigated by fluorescence assay and molecular docking . Under the premise of retaining the structural features, the obtained aptamer was further truncated into a short sequence (named EC1-34), which has the dissociation constant of 17.97 ± 0.98 nM for EC. Subsequently, a visible, rapid, and cost-effective test strip was firstly designed for EC detection in liquor, in which the cationic polymer was used to efficiently capture the recognition probe conjugated with EC1-34 aptamer and gold nanoparticles (AuNPs). The minimum detection limit of the test strip is calculated as 2.14 μg⋅L -1 . The rapid test strip can be widely used for EC detection in liquor due to the advantages of high stability, convenience and selectivity .
研究领域:
分析化学
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纳米技术
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食品科学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Lin Zhang,Siqi Wu,Junfeng Liu,Meiling Ping,Weijuan Yang,FengFu Fu
Sulfonamides (SAs) is a class of antibiotics that extensively used for treating infectious diseases in livestock industries and aquaculture. Thus, it is urgent need to obtain the bio-receptor, which has excellent cross-reactivity and specificity to SAs, for developing high-throughput methods for the determination of multiple SAs even all commonly-used SAs, to realize the quick screening/detection of total SAs in animal-derived foods. We herein isolated several SAs-specific cross-reactive aptamers by using a library-immobilized SELEX with multi-SAs parallel selection strategy. Two of the isolated aptamers (Sul-01 and Sul-04) can specifically recognize and bind seven SAs respectively with higher binding affinity and no interference of non-sulfonamide antibiotics, and thus can be applied as bio-receptors for developing high-throughput aptasensors for the quick screening/detection of multiple SAs. By using the mixture of Sul-01 and Sul-04 as bio-receptor, a ratiometric fluorescent aptasensor was created for the quick detection of nine SAs including sulfamethoxydiazine (SMD), sulfapyridine (SPD), sulfaquinoxaline (SQ), sulfathiazole (ST), sulfamonomethoxine (SMM), sulfamerazine (SMR), sulfaguanidine (SG), sulfamethazine (SMZ) and sulfadiazine (SD) with a detection limit (LOD) of 0.10–0.50 μM, or total of above nine SAs with a LOD of 0.20 μM. The fluorescent aptasensor was successfully applied to detect each or total of SMD, SPD, SQ, ST, SMM, SMR, SG, SMZ and SD in fish samples with a recovery of 83 %–92 % and a relative standard deviation (RSD, n = 5) < 5 %. This study not only provided several promising bio-receptors for the development of diverse high-throughput aptasensors to achieve the quick screening of multiple SAs residues, but also provided a simple, stable and sensitive method for the quick screening of SMD, SPD, SQ, ST, SMM, SMR, SG, SMZ and SD in seafood.
研究领域:
分析化学
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生物技术
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食品科学
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环境科学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2016
·Vol. 183
·期 9
·2555-2562
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IF 4.831
Wu Shijia,Li Qi,Duan Nuo,Ma Haile,Wang Zhouping
Microcystin-RR (MC-RR) is a highly acute hepatotoxin produced by cyanobacteria. It is harmful to both humans and the environment. A novel aptamer was identified by the systemic evolution of ligands by exponential enrichment (SELEX) method as a recognition element for determination of MC-RR in aquatic products. The graphene oxide (GO) SELEX strategy was adopted to generate aptamers with high affinity and specificity. Of the 50 aptamer candidates tested, sequence RR-33 was found to display high affinity and selectivity, with a dissociation constant of 45.7 ± 6.8 nM. Aptamer RR-33 therefore was used as the recognition element in a fluorometric assay that proceeds as follows: (1) Biotinylated aptamer RR-33 is immobilized on the streptavidinylated wells of a microtiterplate, and carboxyfluorescein (FAM) labelled complementary DNA is then allowed to hybridize. (2) After removal of excess (unbound) cDNA, sample containing MC-RR is added and incubated at 37 °C for 2 h. (3) Displaced free cDNA is washed away and fluorescence intensity measured at excitation/emission wavelengths of 490/515 nm. The calibration plot is linear in the 0.20 to 2.5 ng·mL −1 concentration range, and the limit of detection is 80 pg·mL −1 . The results indicate that the GO-SELEX technology is appropriate for the screening of aptamers against small-molecule toxins. The detection scheme was applied to the determination of MC-RR in (spiked) water, mussel and fish and gave recoveries between 91 and 98 %. The method compares favorably to a known ELISA. Conceivably, this kind of assay is applicable to other toxins for which appropriate aptamers are available.
The aptamer RR-33 specific for microcystin (MC-RR) was identified by using the graphene oxide (GO) SELEX process. This aptamer was used for determination of MC-RR by a fluorometric displacement assay.
研究领域:
毒理学
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分析化学
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环境科学
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分子生物学
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产品关联
DOI
高影响因子
ACS Sensors
·
2017
·
IF 0
Shiya Qin,Nandi Chen,Xiaohai Yang,Qing Wang,Kemin Wang,Jin Huang,Jianbo Liu,Maogui Zhou
Pancreatic polypeptide (PP) is a specific biomarker of nonfunctional pancreatic neuroendocrine tumors (NF-pNETs). Clinical significance of PP inspires researchers to make great efforts in developing sensitive and specific sensors. However, there is no existing biosensor for detecting PP that combines facility and functionality. Addressing this challenge, a pair of aptamers which could be used to develop a sandwich assay for PP is reported. First, several high affinity aptamers are screened through graphene oxide-based SELEX, and appropriate dual-aptamers which could bind to different epitopes of PP are identified through fluorescence assays. Then the feasibility of the dual-aptamers for constructing the sandwich assay is validated via dynamic light scattering. This sandwich assay shows considerable sensitivity and specificity. The above results imply that the dual-aptamers have the potential toward developing novel sensors for PP in clinical samples.
研究领域:
生物传感器
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分子生物学
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癌症诊断
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分析化学
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产品关联
DOI
PubMed
Nanoscale
·
2020
·Vol. 12
·期 44
·22574-22585
·
IF 6.895
Tian Gao,Pi Ding,Wenjing Li,Zhili Wang,Qiao Lin,Renjun Pei
Circulating tumor cells (CTCs) acquire mesenchymal markers (e.g., N-cadherin) and lose epithelial markers (e.g., epithelial cell adhesion molecule, EpCAM) during the epithelial-mesenchymal transition (EMT) and are therefore ideal biomarkers of tumor metastasis. However, it is still a challenge to efficiently capture and detect circulating tumor cells with different phenotypes simultaneously. In this work, to obtain aptamers targeting N-cadherin in the native conformation on live cells, we established stable N-cadherin overexpressing cells (N-cadherin cells) and used these cells to identify a panel of N-cadherin-specific aptamers through the cell-SELEX approach. Two aptamer candidates obtained after 12 rounds of selection showed a low equilibrium dissociation constant in the nanomolar range, indicating high binding affinity. The truncated aptamer candidate NC3S showed the highest binding affinity to N-cadherin cells with a low Kd value of 20.08 nM. The SYL3C aptamer was reported to target cancer cell surface biomarker EpCAM. Then, we synthesized two kinds of aptamer-modified magnetic nanoparticles (SYL3C-MNPs and NC3S-MNPs). Both SYL3C and NC3S aptamers possess excellent capture specificity and efficiency for the target cells. The aptamer–MNP cocktail exhibits a considerable capture efficiency and sensitivity for rare cancer cells of epithelial and mesenchymal phenotypes. Furthermore, no CTCs were found in blood samples from healthy donors, while CTCs were successfully isolated by using the aptamer–MNP cocktail for 15 out of 16 samples collected from patients. In summary, the two kinds of aptamer-modified MNPs could be utilized as a promising tool for capturing CTCs from clinical samples.
研究领域:
生物技术
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肿瘤学
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分子生物学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Liping Zhao,Irfan Muhammad,Xiaomin Zhang,Ge Yang,Yao Ma,Bo Wei,Feng Qu
A new apta-sensing probe based on magnetic hydrophilic polymer (MHP) was proposed for simple and sensitive detection of fetuin-A in serum matrix. Briefly, MHP was the hydrophilic polymer modified magnetic nanoparticles , which was synthesized through free radical polymerization method with hydrophilic monomers and magnetic nanoparticles. Then, aptamer against fetuin-A first selected through capillary electrophoresis-SELEX was hybridized with FAM-labeled aptamer complementary sequence (CS) to form duplex structure, which was finally conjugated on MHP to form MHP apta-sensing probe (MHPA). In this probe, MHP with hydrophilic property was employed as separation carrier, aptamer was used for fetuin-A binding and FAM-labeled CS as signal probe . With the presence of fetuin-A, CS escaped and could be detected because of the competitive binding between fetuin-A-aptamer and aptamer-CS. Because of the synergistic effect of specific affinity of aptamer on protein and hydrophilic property of the polymer, MHPA exhibited sensitive and selective detection for fetuin-A. Under optimal conditions, the linear range was 5.0 nM-4.0 μM with the detection limit of 4.4 nM. What’s more, MHPA engaged with good stability and selectivity had been successfully applied in the detection of fetuin-A in serum samples. The as prepared hydrophilic polymer assistant apta-sensing probe with facile and sensitive properties, provides novel explication for efficient protein detection.
研究领域:
分析化学
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生物传感器
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蛋白质检测
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纳米技术
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DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Rong Hu,Xi Zhang,Qiang Xu,Dan-Qing Lu,Yun-Hui Yang,Quan-Qing Xu,Qiong Ruan,Liu-Ting Mo,Xiao-Bing Zhang
A universal aptameric system based on the taking advantage of double-stranded DNA/perylene diimide (dsDNA/PDI) as the signal probe was developed for multiplexed detection of small molecules . Aptamers are single-stranded DNA or RNA oligonucleotides which are selected in vitro by a process known as systematic evolution of ligands by exponential enrichment . In this work, we synthesized a new kind of PDI and reported this aggregated PDI could quench the double-stranded DNA (dsDNA)-labeled fluorophores with a high quenching efficiency. The quenching efficiencies on the fluorescence of FAM, TAMRA and Cy5 could reach to 98.3%±0.9%, 97.2%±0.6% and 98.1%±1.1%, respectively. This broad-spectrum quencher was then adopted to construct a multicolor biosensor via a label-free approach. A structure-switching-triggered enzymatic recycling amplification was employed for signal amplification . High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity towards small analytes . For other targets, changing the corresponding aptamer can achieve the goal. The quencher did not interfere with the catalytic activity of nuclease . The biosensor could be manipulated with similar sensitivity no matter in pre-addition or post-addition manner. Moreover, simultaneous and multiplexed analysis of several small molecules in homogeneous solution was achieved, demonstrating its potential application in the rapid screening of multiple biotargets.
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
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）