Foods
·
2021
·Vol. 10
·期 8
·1822
·
IF 4.35
Qidi Zhang,Ming Zou,Wanyu Wang,Jinyan Li,Xiao Liang
The compound, 4,4′-dinitrocarbanilide (DNC), is the marker residue of concern in edible tissues of broilers fed with diets containing anticoccidial nicarbazin (NIC). In this study, 25 fluorescein-labeled DNC derivatives (tracers) are synthesized and characterized to develop a rapid fluorescence polarization immunoassay (FPIA) for the detection of DNC in chickens using DNC monoclonal antibodies (mAbs). The effect of the tracer structure on the sensitivity of the FPIA is investigated. Our results show that after optimization, the half maximal inhibitory concentrations (IC50) and limit of detection (LOD) of the FPIA in the buffer are 28.3 and 5.7 ng mL−1, respectively. No significant cross-reactivity (CR < 0.89%) with 15 DNC analogues is observed. The developed FPIA is validated for DNC detection in spiked chicken homogenates, and recoveries ranged from 74.2 to 85.8%, with coefficients of variation <8.6%. Moreover, the total time needed for the detection procedure of the FPIA, including sample pretreatment, is <40 min, which has not been achieved in any other immunoassays for DNC from literature. Our results demonstrate that the FPIA developed here is a simple, sensitive, specific, and reproducible screening method for DNC residues in chickens.Keywords:DNC residue;FPIA;tracers;chicken muscle
研究领域:
食品科学
·
免疫学
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产品关联
DOI
PubMed
高影响因子
Foods
·
2023
·Vol. 12
·期 8
·1581
·
IF 5.2
Changfei Duan,Huiyan Zhang,Yingjie Zhang,Qiang Li,Peipei Li,Ghulam Mujtaba Mari,Sergei A. Eremin,Jianzhong Shen,Zhanhui Wang
Erythromycin (ERY) is one of the most common macrolides applied in veterinary medicine to treat diseases or as a feed additive for animal growth promotion. Long-term irrational use of ERY could lead to residues in animal-derived food and the emergence of drug-resistant strains, posing potential threats to human health. In this study, a highly sensitive, specific, robust, and rapid fluorescence polarization immunoassay (FPIA) for the determination of ERY in milk has been described. Herein, to achieve high sensitivity, five tracers of ERY with different fluorescein structures were synthesized and paired with three monoclonal antibodies (mAbs). Under the optimized conditions, the combination of mAb 5B2 and tracer ERM-FITC achieved the lowest IC50value in the FPIA with 7.39 μg/L for ERM. The established FPIA was used to detect ERY in milk, revealing a limit of detection (LOD) of 14.08 μg/L with recoveries of 96.08–107.77% and coefficients of variations (CVs) of 3.41–10.97%. The total detection time of the developed FPIA was less than 5 min from the addition of samples to the result readout. All the above results showed that the proposed FPIA in this study was a rapid, accurate, and simple method for the screening of ERY in milk samples.Keywords:erythromycin;fluorescence polarization immunoassay;homogeneous detection;milk
研究领域:
生物化学
·
食品安全检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2018
·
IF 2.831
Qiyi He,Xiping Cui,Ding Shen,Yingshan Chen,Zhengyun Jiang,Rui Lv,Sergei A. Eremin,Suqing Zhao
In this paper, a simple, rapid and high-throughput fluorescence polarization immunoassay (FPIA) based on polyclonal antibodies (PAb) is described for the determination of glycocholic acid (GCA) in human urine. Three fluorescein-labeled GCA (tracers) with different structures and spacer bridges were synthesized and purified by thin-layer chromatography (TLC). The structure effect of tracers on the assay was investigated and the sensitivity of best tracer in the optimized FPIA demonstrated an IC 50 value of 306 ng/mL. The working range of FPIA was 36 ∼ 2 600 ng/mL and the limit of detection (LOD) was 9 ng/mL. The developed FPIA was time-saving that could be completed within 10 min. Human urine samples spiked with GCA were analyzed by this method, followed by confirmation with commercial enzyme immunoassay analysis (EIA). Excellent recoveries and correlation between these two methods were observed ( R 2 = 0.996), suggesting the developed FPIA could be applied to screening of GCA in human urine samples without complicated cleanup.
研究领域:
医学检验
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2018
·
IF 4.946
Baolei Dong,Sijun Zhao,Hongfang Li,Kai Wen,Yuebin Ke,Jianzhong Shen,Suxia Zhang,Weiming Shi,Zhanhui Wang
In this study, 10 fluorescein-labeled ractopamine (RAC) derivatives (tracers) were synthesized and characterized to develop a rapid fluorescence polarization immunoassay (FPIA) for the detection of RAC in pork, using previously produced RAC polyclonal antibodies. The effect of the tracer structure on the sensitivity of the FPIA was investigated. The specificity of the FPIA was evaluated with 70 β-agonists and β-blockers. The FPIA showed a limit of detection of 0.56 μg kg −1 for RAC in pork, with recoveries ranging from 74.8% to 86.6% in spiked samples. The total analysis time, including sample pretreatment, was less than 1 h. The FPIA was used to screen 150 commercial pork samples for RAC residues and the results were consistent with those of an enzyme-linked immunosorbent assay (ELISA) and ultra-high-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). Our results demonstrate that the FPIA developed here is a rapid, accurate, and sensitive screening method for RAC residues in pork.
研究领域:
食品科学
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免疫学
·
化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF SOILS AND SEDIMENTS
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2017
·Vol. 18
·期 3
·845-851
·
IF 2.522
Guan Tianzhu,Sun Yonghai,Zhang Tiehua,Li Tiezhu,Li Zhuolin,Zhang Yue,Zhang Jie,Wang Yongjun
Purpose Extensive use of bisphenol A (BPA) and its analogues has caused increasing concern over the potential adverse health impacts of these chemicals. The aim of this study was to develop a fluorescence polarization (FP) assay to detect bisphenol A, bisphenol AF (BPAF), tetrabromobisphenol A (TBBPA), and tetrachlorobisphenol A (TCBPA) simultaneously. Materials and methods A recombinant peroxisome proliferator-activated receptor α ligand binding domain (mPPARα-LBD*) was chosen as recognition element in developed FP procedure. The environmental soil samples were assayed for (1) analytical parameters, (2) matrix effect, (3) recovery experiment, and (4) HPLC validation. For statistical analysis, GraphPad Prism 5 software was used. Results and discussion Under the optimized conditions, this assay had an IC 50 of 2.80, 6.84, 4.67, and 4.66 μg g −1 for BPA, BPAF, TBBPA, and TCBPA, respectively. The developed assay demonstrated the limits of detection (LODs) of 0.35, 0.21, 0.27, and 1.03 μg g −1 for these four compounds, which could meet the requirement of environmental monitoring. For spiked soil samples, mean recoveries of the analytes ranged from 85.3 to 101.1%, yielding coefficient of variations (CVs) ranged from 9.8 to 17.5%. Conclusions In this study, a broad-specificity receptor protein for BPA, BPAF, TBBPA, and TCBPA was produced and was used to develop an FP assay in soil samples. The efficiency and simplicity of the proposed assay define its possible application as a screening tool for environmental monitoring.
研究领域:
环境科学
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分析化学
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毒理学
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产品关联
DOI
高影响因子
Analytical Methods
·
2015
·Vol. 7
·期 10
·4246-4251
·
IF 1.821
Pinong Huang,Suqing Zhao,Sergei A. Eremin,Shengwu Zheng,Dan Lai,Yingshan Chen,Bin Guo
Based on a sensitive monoclonal antibody against bisphenol A (BPA) and a new tracer named BVA–AMF, a homogeneous fluorescence polarization immunoassay (FPIA) was developed and applied in the determination of bisphenol A in environmental water samples. BVA was selected as the hapten to couple with bovine serum albumin and the conjugate was used as the immunogen for the monoclonal antibody production. Three fluorescein-labeled BVA tracers with different structures (BVA–AMF, BVA–EDF, and BVA–lysFITC) were synthesized. Under the same optimal conditions, BVA–AMF showed the highest sensitivity for FPIA and the detection of BPA exhibited a limit of detection of 5.60 ng mL−1, an IC50 of 140 ng mL−1 and a dynamic range of 11.32–904.21 ng mL−1 approximately. In this assay, several similar compounds were shown of little significantly with the cross-reactivity being less than 0.15%. Four different kinds of water samples were analyzed, with recoveries being 87.91–114.28%. The detection standard curve of BPA exhibited a good linearity (R2 = 0.9913, n = 3). Compared with ELISA and HPLC methods, FPIA showed reliability and a high correlation with ELISA of 0.9964 and HPLC of 0.9971. The proposed immunoassay technique is suitable for detection of BPA in authentic environmental water samples.
研究领域:
分析化学
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环境科学
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免疫学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Ding Shen,Jie Zheng,Xiping Cui,Yingshan Chen,Qiyi He,Rui Lv,Zhaoxia Li,Suqing Zhao
Trace concentrations of cholyglycine acid (CG) are useful to reflect the damage degree in liver cells, and useful in the diagnosis and prognosis of liver diseases. Therefore, developing an effective strategy to detect the concentration of CG is of great importance. A homogeneous fluorescence polarization immunoassay (FPIA) was developed for determination of CG concentrations. The effects of tracer concentration, antibody dilution and incubation time on FPIA performance were studied. The FPIA was used to detect CG in sera from 522 patients with liver diseases and 449 healthy controls. The results were compared with those obtained through enzyme multiplied immunoassay technique (EMIT). The FPIA showed a detection limit of 50.9 ng/mL for CG and the cross-reactivity of antibodies with nine structurally and functionally related analogs were negligible. Concentrations of CG in patients with chronic hepatitis B virus, liver cirrhosis and decompensated cirrhosis were 5.4, 7.9 and 6.5-fold higher than those in controls, respectively, and hence were significantly different from controls (P < 0.001). The FPIA developed in this study is a rapid, convenient, and simple method, suitable to be used as a screening tool for homogeneous detection of CG in serum. The detection results of FPIA demonstrate that concentrations of CG were much higher in liver patients than in controls. Therefore, CG may be a good biomarker for the diagnosis of liver diseases.
研究领域:
医学检验
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免疫学
·
肝脏疾病研究
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产品关联
DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2022
·
IF 4.831
Wenchao Jia,Xiangying Jin,Yuhua Wu,Danping Xie,Wenhua Yin,Bo Zhao,Zhonghui Huang,Lijun Liu,Yanyan Yang,Tonghui Cao,Xidan Feng,Sheng Chang
Here, we report a novel aptasensor based on decahedral silver nanoparticles (Ag 10 NPs) enhanced fluorescence polarization (FP) for detecting PCB-77. Using aptamer modified Ag 10 NPs hybridized with DNA sequence labeled fluorescent group as an analytical probe, polychlorinated biphenyls (PCB-77) could be detected with high sensitivity and selectivity. The linear range of determination was 0.02 ng/L to 390 ng/L and the limit of detection was 5 pg/L. In addition, through the optimization of the experiment condition and signal probe DNA (pDNA), we found that the maximum FP signal could be generated when the distance between fluorescein and the surface of Ag 10 NPs was 3 nm. When the aptamer was immobilized on the surface of Ag 10 NPs could be strengthened the anti-interference performance of aptamer nanoprobe and further improved the detection ability. At the same time, we also compared the detection performance of the traditional FP signal enhancer streptavidin (SA) analysis system. The fluorescence polarization aptasensor could detect PCB-77 samples efficiently in complex environmental water, which shows a good application prospect.
研究领域:
分析化学
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环境科学
·
纳米技术
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DOI
PubMed
高影响因子
IMMUNOLOGICAL INVESTIGATIONS
·
2012
·
IF 1.164
Xiaoling Wu,Libing Wang,Wei Ma,Yingyue Zhu,Liguang Xu,Hua Kuang,Chuanlai Xu
A rapid, sensitive, specific and simple method based on Fluorescence Polarization Immunoassay (FPIA) was developed for the quantitative detection of Bisphenol A in water. 4, 4-Bis (4-hydroxyphenyl) valeric acid (BHPVA) was selected as the hapten to produce the immunogen. Fluorescein-labeled Bisphenol A derivatives (tracers) with different structures were synthesized and purified by thin layer chromatography. Based on the polyclonal antibody and tracers, an optimized FPIA method was developed with a detection limit (10% inhibition) of 2 μg L−1 and a linear working range of 20 to 800 μg L−1. FPIA was suitable for screening a large number of samples. The recoveries of fortified Tai lake water samples ranged from 91.85 to 102.78%, and tap water samples ranged from 90.36 to 96.01%. The coefficients of variation were all less than 20%. This FPIA method, which did not require a complicated cleanup process, proved to be very useful for the screening of Bisphenol A in environmental water samples.
研究领域:
环境科学
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免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2021
·
IF 4.098
Yingfeng Qin,Gaojie Ye,Hao Liang,Ming Li,Jingjin Zhao
It is highly desirable to develop a simple, efficient and sensitive strategy for organophosphorus pesticides (OPs) in both environment pollution and human health. Herein, a novel amplified fluorescence polarization (FP) biosensor was established for highly sensitive detection of OPs using MnO 2 nanosheets as the signal enhancer. In this system, OPs can suppress the activity of acetylcholinesterase (AChE) efficiently, blocking the hydrolysis reaction of acetylthiocholine (ATCh) to generate thiocholine (TCh) by AChE. TCh can lead the decomposition of MnO 2 nanosheets to manganese ions. So, without the influence of TCh, MnO 2 nanosheets can maintain its original shape and form a stable complex with FAM-DNA, which greatly enhanced the FP signal. This method can tremendously improve the sensitivity of FP with a detection limit of 0.01 ng/mL for diazinon. In addition, it was also applicable to determine other four OPs and investigate the level of diazinon in real water samples. Consequently, the proposed approach provides a new promising platform for detection of OPs and is expected to be used in application of environmental monitoring.
研究领域:
环境科学
·
分析化学
·
生物传感
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ARCHIVES OF TOXICOLOGY
·
2017
·Vol. 92
·期 3
·1215-1223
·
IF 5.901
Zhang Jie,Li Tiezhu,Wang Tuoyi,Yuan Cuiping,Zhong Shuning,Guan Tianzhu,Li Zhuolin,Wang Yongzhi,Yu Hansong,Luo Quan,Wang Yongjun,Zhang Tiehua
The binding interactions of bisphenol A (BPA) and its halogenated derivatives (halogenated BPAs) to human estrogen receptor α ligand binding domain (hERα-LBD) was investigated using a combined in vitro and in silico approach. First, the recombinant hERα-LBD was prepared as a soluble protein in Escherichia coli BL21(DE3)pLysS. A native fluorescent phytoestrogen, coumestrol, was employed as tracer for the fluorescence polarization assay. The results of the in vitro binding assay showed that bisphenol compounds could bind to hERα-LBD as the affinity ligands. All the tested halogenated BPAs exhibited weaker receptor binding than BPA, which might be explained by the steric effect of substituents. Molecular docking studies elucidated that the halogenated BPAs adopted different conformations in the flexible hydrophobic ligand binding pocket (LBP), which is mainly dependent on their distinct halogenation patterns. The compounds with halogen substituents on the phenolic rings and on the bridging alkyl moiety acted as agonists and antagonists for hERα, respectively. Interestingly, all the compounds in the agonist conformation of hERα formed a hydrogen bond with His524, while the compounds in the antagonist conformation formed a hydrogen bond with Thr347. These docking results suggested a pivotal role of His524/Thr347 in maintaining the hERα structure in the biologically active agonist/antagonist conformation. Comparison of the calculated binding energies vs. experimental binding affinities yielded a good correlation, which might be applicable for the structure-based design of novel bisphenol compounds with reduced toxicities and for environmental risk assessment. In addition, based on hERα-LBD as a recognition element, the proposed fluorescence polarization assay may offer an alternative to chromatographic techniques for the multi-residue determination of bisphenol compounds.
研究领域:
生物化学
·
环境科学
·
毒理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Chao Zhang,Tian Tian,Nanzhu Yin,Jingjin Zhao
Ampicillin (AMP) is a β-lactam antibiotic that can inhibit bacterial wall synthesis. The overuse and misuse of AMP makes it micropollutant that commonly found in food and various environmental media. In this work, a fluorescence polarization sensor was designed to sensitive detection of trace ampicillin based on click chemistry, using graphene oxide (GO) as a fluorescence polarization (FP) signal enhancement element. First, when ampicillin binds to its aptamer (apt), the adjacent alkyne and azide groups are separated, hindering the click-linking reaction. When Carboxyfluorescein (FAM) fluorophore-labeled probe (C-FAM) is added, its protruding 3-terminal FAM is recognized and cleaved by exonuclease I (EXO I), releasing fluorophores free that could not be adsorbed on GO, resulting in a lo0wer polarization signal. If there is no AMP in the system, aptamer probe is connected to its complementary chain ends by a click reaction. After C-FAM hybridizes with apt, the apt/P duplex is opened and the prominent single-stranded ends adsorb on the GO, leading a significantly enhanced FP signal. According to the relationship between the difference in FP values and the concentrations of AMP, the limit of detection of proposed method is as low as 80 pg/mL. This assay has a wide linear range plus excellent selectivity, and has been applied to detect AMP in milk and river water samples with satisfactory results, which demonstrates that the FP sensor has great potential for practical applications in food safety and environmental protection fields.
研究领域:
分析化学
·
食品安全
·
环境科学
✓ 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）