MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 9
·1-13
·
IF 5.3
Bi Huaixiu,Zhang Cheng,Zhao Chen,Zhang Xi,Li Chenyu,Wang Shang,Yang Xiaobo,Qiu Zhigang,Wang Jingfeng,Xue Bin,Shen Zhiqiang
The overuse of tetracycline (TC) a novel reverse ELISA assay integrating quantum dots (QDs) with an allosteric transcription factor (TetR) was developed for rapid tetracycline (TC) detection. Specifically, biotin-modified double-stranded DNA (dsDNA) was immobilized on a streptavidin-coated 96-well plate, after which QD-TetR conjugates were added. In the presence of TC, the QD-TetR conjugate binds TC and undergoes allosteric changes that result in its dissociation from the dsDNA. The developed QD-TetR-based reverse ELISA achieved quantitative TC detection within a linear dynamic range of 0.05–100 μM in just 5 min, with a method detection limit of 0.02 μM. The recoveries ranged from 93.43% to 106.6% for actual samples (seawater, sewage, milk, and honey) detection. Furthermore, on-site semi-quantitative detection of TC in actual samples was achieved across a concentration range 0.5–500 μM. The results were validated with commercial ELISA kits, indicating strong potential for monitoring TC residues in environmental and food samples. Graphical abstract
研究领域:
分析化学
·
环境监测
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食品安全
·
生物传感器
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2018
·
IF 2.568
Meirong Song,Zhiming Xiao,Yanan Xue,Xiya Zhang,Shuangyang Ding,Jiancheng Li
An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on immunomagnetic beads' (IMBs) clean-up was developed for detection of the residues of maduramicin (MD) in different chicken tissues. IMBs coated with a specific monoclonal antibody (MAb) against MD named MAb 2D6 were applied to enrich the residues of MD in chicken tissues. The specificities of these IMBs were well maintained and the reversibility remained at more than 73% of the original capability after being used for three times. After elution, enriched MD was detected by a conventional ic-ELISA. The limits of detection of MD were 72, 74 and 173 μg/kg in chicken muscle, skin and fat, and liver, respectively. Recoveries ranged from 80.0% to 115.8% with coefficients of variation being less than 11.3%. These results indicated that a rapid, robust clean-up of IMBs combining ELISA provides a simple, time-saving and environmentally friendly method to detect MD in chicken tissues.
研究领域:
食品科学
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
TALANTA
·
2022
·
IF 6.556
Xueyu Zhang,Mingyan Liu,Minghui Yang,Wangqing Cheng,Juan Xiang,Wu Zhu,Xiang Chen
Nanoparticles are usually used as carrier to load more antibody and enzyme to improve the sensitivity of enzyme-linked immunosorbent assay (ELISA). However, limited by high density and complicated modification procedure, the traditional nanoparticles such as Au nanoparticles (AuNPs) usually induce large background signal and poor reproducibility in ELISA. In this work, functional lightweigh nanoparticle [email protected] ( [email protected] ) was prepared and induced as the carrier of detection antibody and horseradish peroxidase (HRP) to form [email protected] @Ab 2 /HRP biojungates. The appropriate density (close to water) and good hydropilicity ensure the good dispersion of [email protected] @Ab 2 /HRP in solution, preventing the physical sedimention and decreasing the background signal even though the bioconjugate's size is close to 200 nm. The large surface area and abundant active group from PDA facilitate the loading of detection antibody and HRP, improving the loading efficiency and stability of biojungates. Based on it, taking interleukin-17A (IL-17A, a biomarker of psoriasis) as the detection target, we developed a [email protected] sandwich ELISA, achieving a sensitive dynamic range from 0.3 to 80 pg/mL and a detection limit of 0.2 pg/mL. Furthermore, the contents of IL-17A were assayed successfully in 10-fold diluted serum samples from psoriasis patients. Compared with those commercial or AuNP-based ELISA, our [email protected] ELISA method exhibits higher sensitivity, lower background interference, and higher stability, which will significantly improve the application of ELISA in the low-abundant biomolecule assays.
研究领域:
纳米技术
·
生物化学
·
分析化学
·
免疫学
·
诊断学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 8
·1-11
·
IF 5.833
Feng Jun,Lu Hao,Yang Yu,Huang Wenyi,Cheng Hao,Kong Hongxing,Li
Lijun
By in situ synthesis of gold nanoparticles (AuNPs) within the acid-etched (AE) MIL-101 (Cr) framework, AE-MIL-101 (Cr) nanocomposites embedded with AuNPs (AuNP/AE-MIL-101 (Cr)) were prepared as surface-enhanced Raman scattering (SERS) substrate. AuNPs are uniformly distributed and stabilized inside the metal-organic framework (MOF), thus forming more SERS hotspots. The SERS performance of AuNP/AE-MIL-101 (Cr) was evaluated using 4-mercaptophenylboronic acid (4-MPBA), 4-mercaptobenzoic acid (4-MBA), benzidine, and rhodamine 6G (R6G). The SERS substrate displays satisfying stability with very low background signal. When benzidine is used as the Raman reporter, the limit of detection (LOD) can reach 6.7 × 10 −13 mol·L −1 , and the relative standard deviation (RSD) of the intra- and inter-batch repetitive tests is less than 5.2%. On this basis, we developed a method for the detection of human carboxylesterase 1 (hCE 1) in human serum using AuNP/AE-MIL-101 (Cr) nanocomposite as SERS substrate and enzyme-linked immunosorbent assay (ELISA) colorimetric substrate as SERS marker. This method was used to determine hCE 1 in clinical serum samples without complicated sample pretreatment, and the detection results were consistent with the data determined by ELISA. In the concentration range 0.1–120 ng·mL −1 , the SERS signal intensity of benzidine at 1609 cm −1 gradually decreases with the increase of hCE 1 concentration (R 2 = 0.9948). The average recoveries of hCE 1 in human serum are in the range 84 to 108%, with RSDs lower than 7.7%. Graphical abstract
By using AuNP/acid etching-MIL-101(Cr) metal organic framework (MOF) as SERS substrate and enzyme-linked immunosorbent assay (ELISA) colorimetric substrate as the SERS marker, a rapid and sensitive method for the determination of human carboxylesterase 1 (hCE1) in human serum samples has been developed.
研究领域:
分析化学
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Weiguo Wang,Rongkai Du,Changzhi Dong,Jinwu Yan,Lei Zhang
A sensitive enzyme-linked immunosorbent assay (ELISA) has been developed based on the dual catalysis of dendritic Pd-Pt porous nanoparticles and horseradish peroxidase (HRP). The Pd-Pt porous nanoparticles, for the first time, were used to immobilize HRP. As a proof-of-concept demonstration, HRP functioned Pd-Pt porous nanoparticles were applied in hCG detection with a detection limit of 0.07 IU L −1 . The linear intervals is 0.1–6.4 IU L -1 . The Pd-Pt-HRP possesses a better sensitivity than the traditional BA-ELISA. Furthermore, the precision and reproducibility and selectivity were assessed to be eligible for the hCG monitoring.
研究领域:
生物化学
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医学诊断
·
纳米技术
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产品关联
DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Jianing Chen,Zhuoqun Su,Wenrui Li,Ziye Pei,Di Wu,Lin Li,Yongning Wu,Guoliang Li
ABSTRACT Effective detection technologies in food safety with the merits of portable and on-site detection potential are always in pressing demand. Herein, we developed a nanopore-assisted Enzyme-linked immunosorbent assay (NELISA) platform, which innovatively introduced hairpin DNA (HP DNA) probes as reaction substrates. This innovation of substrates effectively avoided the inherent limitations of colorimetric signals (i.e., low sensitivity and inaccurate results) and greatly improved the sensitivity and accuracy of NELISA platform. The alkaline phosphatase (ALP)-modified detection antibody (ALP-Ab2) can specifically bind to ricin and hydrolyze the phosphate groups modified on the HP DNA probes. Nanopore recordings demonstrated that two states of probes produced highly distinguishable nanopore events, enabling the qualitative and quantitative detection of ricin. This NELISA platform fully combined the specificity of ELISA with the ultra-sensitivity, and unique single-molecule fingerprint recognition of nanopore, showing a great on-site detection potential. The established method achieved the ultrasensitive and reliable detection of ricin down to 2.46 fg/mL, which enhanced the detection sensitivity by at least 10 6 -fold compared to traditional ELISA. Furthermore, this proposed method was capable of accurately detecting ricin in real food samples with satisfactory recoveries.
研究领域:
分析化学
·
生物传感器
·
食品安全
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOSCIENCE AND NANOTECHNOLOGY
·
2018
·Vol. 18
·期 2
·951-958
·
IF 1.354
Lou Doudou,Tian Yanyan,Zhang Yu,Yin Junjie,Yang Ting,He Chuan,Ma Ming,Yu Wei,Gu Ning
We found that the peroxidase-like activity of gold nanoparticles (GNPs) followed the Michaelis-Menten kinetic model and was dependent on environmental pH and temperature, which was very similar to natural Horseradish Peroxidase (HRP). However, unlike HRP, which needs a lower H2O2
concentration with a very narrow range to reach a maximum reaction rate and avoid enzyme poisoning, GNPs have very high activity, even at an H2O2 concentration two orders of magnitude higher than HRP. It was demonstrated that H2O2 treatment could
enhance the peroxidase-like activity of GNPs, resulting thus in the activity increase in a circular catalytic reaction by the reduplicative use of GNPs. It was also found that the peroxidase-like activity of GNPs responded sensitively to nanoparticle size and surface modifications. When used
in an immunoassay, GNPs were generally conjugated with antibody and blocked with hydrophilic macromolecules to construct a nanoprobe. This strongly reduced the peroxidase-like activity and detection sensitivity of GNPs, therefore, restricting their use as peroxidase mimetics. We presented
a novel strategy that combined the nanoprobes with gold staining to expose fresh catalytic gold surfaces and obtained a great increase in detection sensitivity.
研究领域:
生物化学
·
纳米技术
·
分析化学
·
免疫学
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DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2021
·
IF 3.935
Xiaomin Pan,Xinxin Xu,Shanshan Song,Liguang Xu,Hua Kuang,Xiaoling Wu,Liqiang Liu,Chuanlai Xu
The compound 2,4-dichlorophenoxyacetic acid (2,4-D) is a synthetic plant growth regulator , which is widely used in agricultural production. Consequently, it is necessary to establish a rapid and sensitive detection method to monitor its use and prevent the environmental and human health problems caused by overuse. In this study, a monoclonal antibody (mAb) 2D5 against 2,4-D was prepared, and based on it, an ic-ELISA and an immunochromatographic strip assay (ICA) were developed for rapid detection. The 50% inhibitory concentration (IC 50 ) of the mAb against 2,4-D was 1.7 ng mL −1 and the mAb showed no cross-reactivity with other plant growth regulators with similar structures to 2,4-D. Under optimum conditions, the ICA showed a visual detection limit (vLOD) of 20 ng mL −1 and a cut-off value of 200 ng mL −1 in bean sprouts, and cabbage samples gave the same results. The quantitative detection showed the linear detection ranges for bean sprouts and cabbage samples were 6.3–73.0 and 5.4–132.6 ng mL −1 , respectively. The ICA was successfully applied to detect 2,4-D in bean sprouts and cabbage samples and achieved good recoveries ranging from 94.7% to 104.3% with a coefficient of variation (CV) less than 8.3%. Hence, the ICA provides a sensitive, efficient and reliable detection method, which has broad application prospects in the detection of a large number of samples.
研究领域:
植物学
·
环境科学
·
分析化学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Hailing Zheng,Hailiang Yang,Yang Zhou,Tianxiao Li,Qinglin Ma,Bing Wang,Qin Fang,Haixiang Chen
The extraction and identification of silk residues in tombs is of great significance for studying the distribution and spread of early silk. However, the complex organic matter in the tomb hinders the accurate identification of silk. In this study, a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) based on immunomagnetic beads (IMBs) was developed for the rapid enrichment and detection of silk residues. The double-antibody sandwich ELISA method established by pairing the IMBs prepared by the silk fibroin monoclonal antibody SF-3 and the silk fibroin monoclonal-labeled antibody bio-SF-1 had the highest detection sensitivity, with a linear detection range of 10 to 104 ng mL–1 and a detection limit of 5.12 ng mL–1. This method was excellent in the extraction and analysis of silk residues from archaeological imprints and soil samples and successfully identified silk residues in samples at the final stage of silk degradation (physical invisible silk). The proteomics analysis results demonstrated the feasibility and practicability of this method.
研究领域:
考古学
·
生物化学
·
材料科学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
·
2019
·
IF 4.527
Zhichang Sun,Xuerou Wang,Zongwen Tang,Qi Chen,Xing Liu
A biotin-streptavidin-amplified enzyme-linked immunosorbent assay using a biotinylated nanobody (BA-Nb ELISA) was developed to detect ochratoxin A (OTA) in cereal. The limit of detection (LOD) of the BA-Nb ELISA, which equals to 10% maximal inhibitory concentration, was 0.011 ng/mL for OTA in buffer, and the sensitivity was approximately improved by one order of magnitude compared with the traditional Nb ELISA (LOD = 0.112 ng/mL). Under optimal conditions, the developed assay could be accomplished in 40 min with maximal inhibitory concentration of 0.138 ng/mL and the linear detection range of 0.034–0.460 ng/mL. The average recovery rate of the BA-Nb ELISA ranged from 92.8% to 114%, and the relative standard deviation was in the range of 2.04–9.85%. The developed BA-Nb ELISA was validated by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and the results indicated the reliability of BA-Nb ELISA for the detection of OTA in cereal.
研究领域:
分析化学
·
食品科学
·
毒理学
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2015
·Vol. 7
·期 13
·5635-5640
·
IF 1.821
Yajing Lei,Xiaoyan Yao,Dai Zhang,Lifang Zhou,Weixian Lu,Shuqing Chen
Salbutamol (SAL) is used to promote animal growth, because it increases feeding efficiency and carcass lean percentage. SAL has been banned in many countries because of the potential hazard to human health. To investigate the SAL residue level in humans, a sensitive direct competitive ELISA was developed. Under the optimal experimental conditions, the limit of detection (LOD) for the assay was 0.01 ng mL−1, and the standard curve was constructed at concentrations of 0.01–100 ng mL−1. We used the developed assay to detect the urinary concentration of SAL in 1648 participants. SAL was detected in 96% of participants with a concentration ranging from the LOD to 30.83 ng mL−1. Pregnant women were found to have a lower SAL concentration than non-pregnant women. Sources of exposure, risk assessment and measures for reducing the exposure level require further investigation.
研究领域:
分析化学
·
毒理学
·
公共卫生
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2019
·
IF 8.456
Huaping Peng,Zhongnan Huang,Weihua Wu,Mingkai Liu,Kaiyuan Huang,Yu Yang,Haohua Deng,Xinghua Xia,Wei Chen
This report outlines a versatile high-performance electrochemiluminescence (ECL) enzyme-linked immunosorbent assay (ELISA) platform, which combines the merits of high-quantum-yield Au nanocluster (AuNC) probe-based ECL technology, the efficient ECL-resonance energy-transfer (ECL-RET) strategy, and highly sensitive and specific ELISA technology. The ECL detection procedure was performed on a recyclable MnO2/AuNC-modified glassy carbon electrode interface by taking advantage of the ECL-RET between the AuNC probe and MnO2 nanomaterials (NMs) to quench the ECL intensity. The etching of MnO2 NMs by the product of ALP-based ELISA recovers the ECL signal. Notably, the ELISA process and the ECL detection procedure in this system are independent. Thus, the ECL-ELISA system can effectively avoid the influence of complex biological samples, and the ECL efficiency of the AuNC probe can be used readily. As demonstrated on TNF-α, because of the abovementioned characteristics, the ECL-ELISA platform presented an extremely wide dynamic range, with a detection limit of 2 orders lower than ELISA. Moreover, the system was also applicable for ultrahigh sensitive detection of various disease-related proteins and able to detect trace biomarkers in real serum samples. Therefore, this multifunctional ECL assay platform is versatile, facile, ultrasensitive, recyclable, and sufficiently straightforward for trace biomarker detection in complex biological samples. This approach not only enriches the foundational study of ECL devices but also greatly expands the potential application of ECL sensors in biological testing and clinical high-throughput diagnosis.
研究领域:
分析化学
·
生物化学
·
生物技术
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）