Analytical Methods
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2014
·Vol. 6
·期 7
·2306-2313
·
IF 1.938
Jianhua Shu,Liming He,Huanzhong Ding,Liqi Wang,Hongyan Guo,Yan Gao,Emmanuel E. Dzakah,Zhenling Zeng
A sensitive and specific monoclonal enzyme-linked immunosorbent assay (ELISA) for the determination of tissue-bound metabolite 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ) was developed. Three different haptens of AMOZ were synthesized. The haptens (I, III) were derived from 4-carboxybenzaldehyde and ethyl 4-bromobutyrate, respectively. Hapten II was based on hapten I but a CN bond in the parent structure was reduced. Corresponding immunogens and coating antigens were prepared to improve the assay sensitivity. The hybridomas 4 × 108 secreting antibodies against CPAMOZ were obtained from immunogens I-BSA by monoclonal antibody (mAb) technology. The antibody indicated that: (1) the small molecular AMOZ cannot induce the specific response against AMOZ; (2) the weakened conjugation effect improved the recognition of AMOZ. Assessment of twelve coating antigen/antibody combinations consequently resulted in the development of an indirect competitive enzyme-linked immunosorbent assay (icELISA). The results showed that the sensitivity was highly improved about fifteen-fold for the novel heterologous coating antigen at low hapten density. The IC50 value of the ELISA for CPAMOZ was 0.13 ng mL−1. The cross reactivity values of the assay with NPAMOZ, AMOZ and FTD were 118%, 2.3% and 16.3%. Recoveries from AMOZ fortified animal tissues were in a range of 82.6–108.4% and the CV values were less than 12.5%. The immunoassay was further validated by a LC-MS/MS method and the two methods showed good correlation (r2 = 0.9908). The proposed icELISA could be used for an accurate monitoring of AMOZ in animal tissues.
研究领域:
分析化学
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免疫学
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兽医学
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DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
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2018
·
IF 2.568
Fengyin Liu,Zijian Chen,Yudong Shen,Yuanming Sun,Jinyi Yang,Hong Wang,Hongtao Lei,Zhenlin Xu
Immunoassay direct for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ), the metabolite of furaltadone, based on a specific antibody was usually unavailable due to its small molecule weight. In this work, a hapten with a double bond and an active carboxyl group was designed and then conjugated to peptide dendrimer to obtain novel multiple haptens. The haptens and multiple haptens were coupled to bovine serum albumin as immunogens. Polyclonal antibodies showed binding to free AMOZ in a heterologous competitive indirect enzyme-linked immunosorbent assay were obtained. The assay showed IC50 (half maximal inhibitory concentration) of 4.1 μg/kg and limit of detection of 0.2 μg/kg for AMOZ. Recoveries of AMOZ from grass carp were tested from 83.1% to 117.0%, with the coefficient of variation below 12%. As no laborious derivatisation procedure was employed, the proposed assay should be useful for the screening of AMOZ for a large number of samples.
研究领域:
生物化学
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免疫学
·
食品科学
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2015
·
IF 4.097
Mingxin Li,Hong Yang,Shuqun Li,Chunwei Liu,Kang Zhao,Jianguo Li,Danni Jiang,Lulu Sun,Hong Wang,Anping Deng
3-Amino-5-methylmorpholino-2-oxazolidinone (AMOZ) is the metabolite of furaltadone (FTD), one kind of nitrofuran antibiotics which have been banned as veterinary drugs due to the potential harmful effects on human health. AMOZ is stable in tissue matrices, and can be used as an indicator for illegal usage of FTD. In this study, a novel ultrasensitive competitive immunochromatographic assay (ICA) based on surface-enhanced Raman scattering (SERS) for direct detection of AMOZ in tissue and urine samples was developed. Monoclonal antibody (mAb) against AMOZ and Raman reporter 4-mercaptobenzoic acid (MBA) dual labelled colloidal GNPs ( e.g. Ab-Au-MBA) was prepared, characterized and used as an immunoprobe. The specific Raman scattering intensity of MBA on the test line of the ICA strip was measured for quantitative determination of AMOZ. The assay was completed within 15 min. The values of IC 50 and limit of detection (LOD) of the assay for AMOZ were 0.04 ng mL −1 and 0.28 pg mL −1 , respectively. There was no cross-reactivity (CR) of the assay with other three nitrofurans and five compounds. The ICA strips could be used for 7 weeks without loss of activity. The tissue and urine samples were spiked with AMOZ and the recoveries of AMOZ were in range of 87.8–105.9% with the relative standard deviations (RSD) of 1.4–9.0% ( n = 3). The results demonstrated that the proposed SERS-based ICA was able to directly detect AMOZ with high sensitivity, specificity, stability, precision and accuracy, and might be as a powerful method for the analysis of other analytes in different matrices.
研究领域:
分析化学
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兽医学
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食品安全检测
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2016
·
IF 2.857
Feng Wang,Hong Wang,Yu-Dong Shen,Yong-Jun Li,Jie-Xian Dong,Zhen-Lin Xu,Jin-Yi Yang,Yuan-Ming Sun,Zhi-Li Xiao
A new multianalyte immunoassay was designed to screen furaltadone metabolite 5-morpholinomethyl-3-amino-2-oxazolidone (AMOZ), malachite green (MG), and leucomalachite green (LMG) in aquatic products using a bispecific monoclonal antibody (BsMAb). Gradient drug mutagenesis methods were separately used to prepare an anti-3-nitrobenzaldehyde-derivatized AMOZ (3-NPAMOZ) hybridoma cell line that was hypoxanthine-guanine-phosphoribosyltransferase (HGRPT) deficient and an anti-LMG hybridoma cell line that was thymidine kinase (TK) deficient. BsMAb recognizing 3-NPAMOZ and LMG was generated using hybrid–hybridomas of HGRPT and TK deficient cell lines. For AMOZ and LMG, respectively, the BsMAb-based indirect competitive ELSIA (ic-ELISA) values of 1.7 ng/mL and 45.3 ng/mL and detection limits of 0.2 ng/mL and 4.8 ng/mL. To establish the ic-ELISA, 3-NPAMOZ derivatized from AMOZ with 3-nitrobenzaldehyde and LMG reduced from MG by potassium borohydride was recognized by BsMAb. Recoveries of AMOZ, MG, and LMG in aquatic products were satisfactory and correlated with HPLC analysis. Thus, the multianalyte ic-ELISA is suitable for rapid quantification of AMOZ, MG, and LMG in aquatic products.
研究领域:
分析化学
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食品科学
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免疫学
·
水产养殖
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产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2021
·
IF 3.101
Xinzhu Chen,Huaying Du,Yanping Hong,Yubo Wang,Mei Shu,Dan Wang,Peng Dai,Weijie Deng,Jianhua Xiong,Wuying Yang
To determine furaltadone metabolites 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) residual levels in ctenopharyngodon idellus, direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. Firstly, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against AMOZ derivative, and then the productive VH and VL genes were assembled to a scFv gene and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step dcELISA against AMOZ derivative was developed with a half-maximal inhibitory concentration (IC50) and limit of detection (LOD) of 10.62 and 4.83 ng/mL, respectively. The recovery values of AMOZ from spiked fish samples determined by dcELISA were in 71.31%∼82.88%. The results suggested that the prepared scFv-AP fusion protein in this work can be used for rapid and convenient immunoassays to detect AMOZ residues in fish samples.
研究领域:
免疫学
·
生物化学
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分析化学
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水产养殖
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食品安全
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2026
·Vol. 193
·期 4
·272
·
IF 5.4
Yang Dong,Li Xiangyu,Wu Jing,Yao Tuanli,Chen Chaoxian,Li Xuhua,Chen Jifan,Lan Xueyun,Wang Jiao,Wang Haihua
Semicarbazide (SEM), a metabolite of nitrofurazone, poses potential risks as a carcinogen and teratogen in animal-derived foods. This research is centered on creating a homemade electrochemical sensor with high sensitivity for detecting SEM with dual signals. The working electrode was modified through the in-situ growth of a copper-iron bimetallic organic framework (Cu/Fe-BTC) on carbon cloth (CC), followed by the decoration of gold nanoparticles (AuNPs) on the Cu/Fe-BTC/CC surface. The Cu/Fe-BTC component provides abundant metallic active sites, resulting in a distinct [Fe(CN) 6 ] 3−/4− peak observed at 0.67 V (I 1 , vs. SCE). Concurrently, the incorporation of AuNPs enhances the electrode high electron transfer capacity, which amplifies the signal peaked at 0.18 V (I 2 , vs. SCE), thereby establishing an effective dual-signal detection system. Furthermore, a molecularly imprinted polymer (MIP) layer was electropolymerized onto the modified electrodes, utilizing resorcinol as the functional monomer and SEM as the template. Then, the peak intensities of the dual signals decreased upon target (SEM) recognition and increased during template elution, generating a measurable detection signal. By calculating the current changes, ΔI D = ΔI 1 + ΔI 2 , the adsorbed template molecule concentration could be quantified. Under optimal conditions, the sensor has excellent stability, accuracy, and repeatability in analyzing various samples, including serum, urine, meat, egg, honey, canned food, flour, milk powder, fish, and shrimp. The determination range is linear, spanning from 0.005 to 100 nM, with a limit of detection at 3.9 pM. Therefore, the Cu/Fe-BTC and AuNPs-based MIP sensor represents a reliable dual-signal strategy for the quantitative determination of SEM. Graphical
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PubMed
高影响因子
ChemistrySelect
·
2026
·Vol. 11
·期 8
·e07116
·
IF 2.3
Xiongtao Yu,Haiping Xu,Rui Wang,Feng Zhou,Chunmin Yang
In this manuscript, Au nanoparticles (AuNPs) and a nanospherical conjugated microporous polymer (CMP) were prepared and then modified on the surface of screen-printed carbon electrode (SPCE) to construct an electrochemical sensor for high sensitivity and rapid detection of 3-amino-2-oxazolidinone (AOZ). The preparation process of the materials and electrodes were characterized by scanning electron microscope (SEM), transmission electron microscope (TEM), cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance (EIS), and the test conditions were optimized. Under optimal conditions, the oxidation peak current of AOZ and the concentrations show a wider linear relationship in the ranges of 1.0×10 −8 ∼1.0×10 −6 and 1.0×10 −6 ∼ 1.0×10 −4 mol/L ( R 1 2 = 0.996, R 2 2 = 0.998), respectively, and a lower detection limit which is as low as 3.3×10 −9 mol/L (S/N = 3) compared to existing methods. The reason for this result may be that CMP exhibited abundant porous structures and numerous active sites, which could significantly improve response sensitivity; AuNPs also possess high specific surface area and make the diffusion and transport of electrolyte ions easier. CMP-AuNPs nanocomposites greatly improved the electron transport rate, signal intensity and electroactive surface of SPCE, thereby improving the sensitivity of the sensor. It is worth noting that the sensor also has strong anti-interference ability, stability, and reproducibility. When applied to detect AOZ in real samples, it also showed good recovery rate. Compared with HPLC-MS/MS, the sensor also obtained good accuracy and consistency, which has great application potential for rapid on-site analysis and detection in animal-derived food not only for AOZ but also for Furazolidone (FZD).
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高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Xu Wen,Xueling Yan,Xiaoling Zheng,Qiming Kou,Lulan Yang,Jiaming Tang,Xingyue Chen,Yong Xie,Tao Le
Nitrofurazone (NFZ) is an antibiotic banned in many countries, as its residue seriously harms the human body. Herein, anti-NFZ aptamers were selected and identified based on the magnetic bead SELEX technique using a ssDNA library with a full length of 90 nucleotides (nt). Five full sequence candidate aptamers (NFZ8, NFZ24, NFZ28, NFZ34, and NFZ70) were obtained by secondary structure analysis. We optimized the entire sequence to obtain a truncated aptamer, a 16 nt sequence (NFZ8-1:5′-GTTCTATTGAAAAAAC-3′) that showed the highest affinity for NFZ (K d = 76.11 nM). The binding site of NFZ and aptamer NFZ8-1 was found to be "GAA" by molecular docking . In addition, utilizing the most special truncated aptamer NFZ8-1 as the identification probe, a graphene oxide fluorescent aptasensor is an innovative for the detection of NFZ residue that showed a wide linear reach from 1.25 to 160 ng/mL and a low limit of detection of 1.13 ng/mL. In the actual water environment sample detection, the recovery rate ranged from 95.21 to 113.66%, and the coefficient of variation ranged from 3.53 to 11.24%. These results demonstrate that the NFZ-truncated aptamer applied to the aptasensor provides a novel methodology for recognizing NFZ residues.
研究领域:
生物化学
·
分析化学
·
药物检测
✓ 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）