MOLECULES
·
2023
·Vol. 28
·期 15
·5743
·
IF 4.6
Yexuan Ren,Ruwen Tian,Ting Wang,Junlin Cao,Jianguo Li,Anping Deng
In this study, an extremely highly sensitive enzyme-linked immunosorbent assay (ELISA) based on a newly produced monoclonal antibody (mAb) for the detection of ochratoxin A (OTA) in food samples was developed. OTA-Bovine serum albumin (BSA) conjugate was prepared and used as the immunogen for the production of the mAb. Among four hybridoma clones (8B10, 5C2, 9B7, and 5E11), the antibody from 8B10 displayed the highest affinity recognition for OTA. Based on the mAb (8B10), the IC50and LOD of the ELISA for OTA were 34.8 pg mL−1and 1.5 pg mL−1, respectively, which was 1.53~147 times lower than those in published ELISAs, indicating the ultra-sensitivity of our assay. There was no cross-reactivity of the mAb with the other four mycotoxins (AFB1, ZEN, DON, and T-2). Due to the high similarity in molecular structures among OTA, ochratoxin B (OTB), and ochratoxin C (OTC), the CR values of the mAb with OTB and OTC were 96.67% and 22.02%, respectively. Taking this advantage, the ELISA may be able to evaluate total ochratoxin levels in food samples. The recoveries of the ELISA for OTA in spiked samples (corn, wheat, and feed) were 96.5–110.8%, 89.5–94.4%, and 91.8–113.3%; and the RSDs were 5.2–13.6%, 8.2–13.0%, and 7.7–13.7% (n= 3), respectively. The spiked food samples (corn) were measured by ELISA and HPLC-FLD simultaneously. A good correlation between ELISA (x) and HPLC-FLD (y) with the linear regression equation of y = 0.918x − 0.034 (R2= 0.985,n= 5) was obtained. These results demonstrated that the newly produced mAb-based ELISA was a feasible and ultra-sensitive analytical method for the detection of OTA in food samples.Keywords:enzyme-linked immunosorbent assay (ELISA);monoclonal antibody (mAb);mycotoxin;ochratoxin A (OTA);food samples;sensitive detection
研究领域:
食品科学
·
免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMICAL RESEARCH IN CHINESE UNIVERSITIES
·
2017
·Vol. 33
·期 6
·876-881
·
IF 1.024
Su Ping,Chen Xiaonan,He Zhangjing,Yang Yi
Kanamycin is an aminoglycoside antibiotic used increasingly in human and veterinary medicine. However, kanamycin residues in food can cause serious side effects. Here we reported the preparation of polyclonal antibody and the development of an indirect competitive biotin-streptavidin-amplified-based enzyme-linked immunosorbent assay(BA-ELISA) that can sensitively and specifically detect kanamycin residues in milk and honey. The immunogen and coating antigen were synthesized by covalently linking kanamycin to carrier proteins using the carbodiimide method. The anti-kanamycin polyclonal antibodies were obtained from immunized rabbits. The key assay parameters were investigated and optimized. The results show that under optimum conditions, the limit of detection for kanamycin is 0.07 ng/mL and the IC 50 is 6.48 ng/mL. Cross-reactivity values of the antibody with four kanamycin analogues are all <1%. Trace amounts of kanamycin in milk and honey samples can be detected by this novel BA-ELISA method successfully with satisfactory recoveries of 91.0%—103.3%. The developed protocol was also validated against liquid chromatography-mass spectrometry, returning a significant correlation. These results indicate that BA-ELISA is a viable option for monitoring kanamycin residues in milk and honey.
研究领域:
免疫学
·
分析化学
·
食品科学
·
兽医学
✓ 已验证
产品关联
DOI
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Nuanfei Zhu,Xuesong Li,Ye Liu,Jingfu Liu,Yawei Wang,Xiangyang Wu,Zhen Zhang
Dibutyl phthalate (DBP) is considered as one of the most widely used phthalate esters (PAEs), which has attracted worldwide concerns because of its potential threats to eco-environments and human health. Systematic investigations of DBP environmental occurrence contribute to the further risk assessment, which depends on effective and available analytical methods. In this study, an amplified ratiometric fluorescence ELISA was established for sensitive and high-throughput detection of DBP in the aquatic system based on a novel tetrahedral DNA nanostructure (TDN)-scaffolded-DNAzyme (Tetrazyme). Wherein, Tetrazyme was prepared by the precise folding of G-quadruplex sequence on three vertex angles of the TDN, together with hemin as the horseradish peroxidase (HRP)-mimicking enzyme. The rigid TDN avoided the local overcrowding effect to provide a reasonable spatial spacing on the interface for G-quadruplex sequence, increasing the collision chance between DNAzyme and substrates, improving the catalytic ability of DNAzyme effectively. Besides, streptavidin (SA) and biotin (bio) were used to anchor TDN and antibody, in which the specific binding of SA/bio could make more Tetrazyme conjugate on each signal element, resulting in the dual signal amplification. Meanwhile, the accuracy and precision were enhanced owing to the inherent built-in rectification to the environment from the dual output ratiometric fluorescence assay. Under the optimized conditions, the detection limit of this proposed method was 0.17 ng/mL (16 times lower than that of conventional ELISA using the same antibody) with a satisfactory accuracy (recoveries, 79.0%- 116.2%; CV, 2.1–6.5%). Overall, this platform provides a promising way for accurate, sensitive and rapid determination of DBP from environmental waters.
研究领域:
环境科学
·
分析化学
·
纳米技术
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Liyan Zhang,Chuan Fan,Min Liu,Fengjuan Liu,Shanshan Bian,Shuyue Du,Shuyun Zhu,Hua Wang
Catalytic hemin was first remolded with biomineralized gold to form Hemin-Au core and further encapsulated into the Tb metal organic framework (MOF) matrix. The so obtained Hemin-Au@MOF composites could possess high environmental stability and especially double catalysis activities of enhanced peroxidase-like and gold catalysis. Steady state kinetics studies indicate that Hemin-Au@MOF could present much higher intrinsic catalysis and substrate affinity (lower Michaelis constants) than native hemin and even natural peroxidase . Furthermore, the catalytic composites were employed for labeling antibodies towards the sandwiched immunoassays for alpha-fetoprotein (AFP) as a model biomarker of cancers. The results indicate that the Hemin-Au@MOF-based immunoassays could be conducted by two signal amplification ways of hemin-catalyzed chromogenic reaction and gold-catalyzed silver staining, enabling the detection of AFP in blood with the level down to 0.020 ng mL −1 . Importantly, the proposed fabrication protocol combining the gold biomineralization and MOF encapsulation routes may be extended for the fabrication of a variety of enzymes or enzymatic mimics with the improved environmental stability and multiple catalysis activities for the wide catalysis applications.
研究领域:
生物化学
·
医学诊断
·
材料科学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2023
·Vol. 415
·期 10
·1877-1887
·
IF 4.3
Wang Yinghui,Yu Ling,Zhang Hui,Zhu Runzhi,Meng Zhe
An accurate and sensitive competitive enzyme-linked immunosorbent assay (ELISA) based on persistent luminescence nanoparticles Zn 2 GeO 4 :Mn 2+ , Eu 3+ (ZGME) was developed for detecting ochratoxin A (OTA), a powerfully toxic mycotoxin usually found in grains. As a signal output element of autofluorescence-free biosensors, ZGME can be integrated into ELISA with glucose oxidase (GOx)-binding OTA molecules due to its excellent pH-responsive persistent luminescence. In the absence of OTA, the OTA-GOx conjugate was captured by the anti-OTA monoclonal antibody (anti-OTA mAb) pre-coated on the 96-well plate. The results indicate a decrease in the pH value of the solution, which triggered the quenching of ZGME luminescence due to GOx-dependent gluconic acid production. The presence of OTA inhibited the binding of OTA-GOx on the plate, thus decreasing the production of gluconic acid and increasing the persistent luminous intensity of ZGME. Under the optimized concentrations of anti-OTA mAb and OTA-GOx, quantitative determination of OTA was achieved by plotting the increase or decrease in persistent luminescence intensity of ZGME at 535 nm. In this study, the linear range was from 0.1 μg L −1 to 63 μg L −1 , and the limit of detection (LOD) was as low as 0.045 μg L −1 . In five food samples (corn grit, brown rice, soybean, rice, and wheat), the results exhibited good stability and repeatability, with a recovery range from 81.3% to 94.4% and a relative standard deviation (RSD) of less than 4.2%. Hence, the established method provides a sensitive, accurate, and autofluorescence-free approach for the determination of OTA in different grain samples.
研究领域:
分析化学
·
毒理学
·
食品科学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2013
·Vol. 5
·期 21
·6106-6113
·
IF 1.855
Yajing Lei,Lizheng Fang,Muhammad Sajid Hamid Akash,Zhiming Liu,Weixing Shi,Shuqing Chen
Bisphenol A (BPA) is widely used to manufacture polycarbonate plastics and epoxy resins, which are in widespread use in China. Concerns about potential health impacts from exposure to BPA among the general population are increasing day by day. A sensitive and specific antibody was prepared to develop and compare direct and indirect competitive enzyme-linked immunosorbent assays (ELISAs) for the determination of BPA in human urine. The IC50 value and limit of detection (LOD) for the direct competitive ELISA were 5.5 ng mL−1 and 0.03 ng mL−1 respectively whereas, for the indirect competitive ELISA, the IC50 value and LOD were 7.0 ng mL−1and 0.08 ng mL−1, respectively. We found that the direct competitive ELISA was more specific and sensitive than the indirect competitive ELISA. The established immunoassays were also applied in the determination of BPA in human urine samples. The results from fortified samples and real samples indicated that the direct competitive ELISA was more sensitive and reliable for the measurement of BPA in human urine as compared to the indirect competitive ELISA.
研究领域:
环境健康
·
分析化学
·
毒理学
✓ 已验证
产品关联
DOI
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2014
·Vol. 38
·期 7
·2935-2940
·
IF 3.159
Lei Zhan,Wen Bi Wu,Xiao Xi Yang,Cheng Zhi Huang
Respiratory syncytial virus (RSV) mediates severe respiratory illness in infants and young children worldwide, and it is also a significant pathogen of the elderly and immune compromised. Due to the low infectious dose and high infectious rate of RSV, remarkably rapid and sensitive RSV detection is important for infection control and discovery of antiviral drugs. In this work, we described a highly sensitive enzyme-linked immunosorbent assay (ELISA) utilizing gold nanoparticles (AuNPs) for the detection of RSV. Gold nanoparticles were used as carriers of the signalling antibody anti-RSV–HRP (horseradish peroxidase) in order to achieve an amplification of the signal. Compared to conventional ELISA procedures, this assay resulted in higher sensitivity and shorter assay time in the range between 0.5 and 50 pg mL−1. The application of AuNPs to the enhanced ELISA showed acceptable reproducibility, stability, and could be readily applied to determination of other pathogens in clinical diagnostics.
研究领域:
病毒学
·
纳米技术
·
临床诊断
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
International Journal of Environmental Research and Public Health
·
2013
·Vol. 10
·期 4
·1598-1608
·
IF 1.998
Hua Kuang,Wenbing Wang,Liguang Xu,Wei Ma,Liqiang Liu,Libing Wang,Chuanlai Xu
A sensitive and specific monoclonal antibody-based sandwich enzyme-linked immunosorbent assay (ELISA) was established and validated for the detection of staphylococcal enterotoxin A (SEA). After routine fusion and selection, 10 monoclonal antibodies showed high affinity for SEA. An optimal pair for sandwich ELISA was selected by pairwise interaction analysis. After optimization, the limit of detection (LOD) and linear dynamic range of the method were established, and were found to be 0.0282 ng/mL and 0.06–2 ng/mL, respectively. The recovery in pure milk ranged from 82.67% to 111.95% and the intra- and inter-assay coefficients of variation ranged from 3.16% to 6.05% and from 5.16% to 10.79%, respectively. Cross-reactivity with staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), staphylococcal enterotoxin D (SED), and staphylococcal enterotoxin E (SEE) in this method were insignificant. These results indicate that the sandwich ELISA method developed in our study is effective for routine identification of SEA in food samples.
研究领域:
食品科学
·
免疫学
✓ 已验证
DOI
高影响因子
FOOD CHEMISTRY
·
2017
·
IF 4.529
Lu Li,Ai-hong Peng,Zheng-zhong Lin,Hui-ping Zhong,Xiao-mei Chen,Zhi-yong Huang
A highly selective and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of malachite green (MG) using a molecularly imprinted polymer (MIP) film as bionic antibody. The MIP film, based on the self-polymerization of dopamine, was fabricated on the surfaces of a 96-well microplate. It showed specific recognition for MG in aqueous solution. A direct competitive ELISA method was established with the sensitivity reaching 10.31 μg L −1 and the detection limit being 0.3 μg L −1 . The cross-reactivity of two structural analogues to MG was less than 10%. The average recovery tested by MG standard spiking was 88.8% for bass and 90.4% for water, and the relative standard deviations were less than 3.6%. All the above results indicated that the developed method could be used to detect MG in fish and water samples rapidly, specifically and accurately.
研究领域:
分析化学
·
环境科学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2011
·
IF 0.633
Fei Lin,Shanshan Song,Liqiang Liu,Hua Kuang,Libing Wang,Chuanlai Xu
An enzyme-linked immunosorbent assay method was developed here for benzophenone (BZP) detection in package paper samples based on polyclonal antibody. 4-Aminobenzophenone (ABZP) was used as hapten and conjugated with bovine serum albumin for antibody production. Cross-reactivity results showed that the antibody can be recognised BZP, ABZP and Michler's ketone very well (IC50 value was 31.2±7.8 ng/mL, 35.9±8.3 ng/mL and 43.7±8.2 ng/mL, respectively). After optimisation of the effects from pH, ionic strength and organic solvents on immunoassay, we established a rapid method and used for BZP detection in fortified package paper samples with good recovery data (98.5 and 99.7%) at 100 and 600 ng/g spiked level.)
研究领域:
环境科学
·
分析化学
✓ 已验证
DOI
高影响因子
FOOD CHEMISTRY
·
2021
·
IF 7.514
Jing Zhao,Yaohai Zhang,Qiyang Zhao,Yue He,Zhixia Li,Aihua Chen,Chengqiu Wang,Baomin Wang,Bining Jiao,Yongliang Cui
Naringenin, a flavonoid compound found in pummelo, is a key biological active compound in some traditional Chinese medicines, including Citri reticulatae pericarpium , Citri reticulatae pericarpium viride , Aurantii fructus immaturus , and Aurantii fructus . These Chinese medicinal preparations are the peels or immature fruits of certain citrus species. Aiming at detecting naringenin in complex matrices such as pummelo and traditional Chinese medicines, we put forward a sensitive and practical indirect competitive enzyme-linked immunosorbent assay (icELISA) based on anti-naringenin monoclonal antibodies (anti-Nar-mAbs). The median inhibitory concentration (IC 50 ) was 4.43 ng/mL, and the working range was 1.15–15.81 ng/mL. The findings of the icELISA for the analysis of naringenin in pummelo and herb samples had a good correlation with the ultra performance liquid chromatography (UPLC) methodology and showed good accuracy and reproducibility. These data demonstrated that the developed icELISA is reliable, accurate, and suitable for detecting naringenin in pummelo and traditional Chinese medicines.
研究领域:
药理学
·
分析化学
·
中药学
·
食品科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2021
·
IF 7.514
Sihan Wang,Zile Wang,Liang Zhang,Yuliang Xu,Jincheng Xiong,Huixia Zhang,Zhiwei He,Yongjun Zheng,Haiyang Jiang,Jianzhong Shen
Metal-organic frameworks (MOFs) has excellent adsorption performance, herein, three kinds of common MOFs were used for the adsorption of sulfamethazine (SM 2 ) in milk, then enzyme-linked immunosorbent assay (MOF-ELISA) was established. Firstly, NH 2 -UiO-66, NH 2 -MIL-101, and ZIF-8 were successfully prepared and their adsorption characteristics for SM 2 were investigated. The kinetic models of the three MOFs were more in line with the pseudo-second-order adsorption kinetics, and the saturated adsorption capacity of NH 2 -UiO-66, NH 2 -MIL-101, and ZIF-8 for SM 2 at 298 K were 139.64, 29.98, and 36.5 mg/g, respectively. Using three different MOFs as adsorbents, the pretreatment of milk samples could be completed within 1 h, the half inhibitory concentrations (IC 50 ) of MOF-ELISA were 1.26, 1.86 and 2.74 ng/mL, the limit of detections (LOD) were 0.05, 0.12, and 0.19 ng/mL and the recovery rate were from 82.30% to 105.62% with the intra-day coefficient of variations (CVs) below 5.81% and inter-day CVs below 7.21%. Detection results showed good correlations with LC-MS/MS (R 2 > 0.99), indicated that MOFs could effectively eliminate the interference of sample matrix, and has the potential to become a general pretreatment method for the detection of various matrices residues in food safety monitoring.
研究领域:
食品安全
·
分析化学
·
生物技术
✓ 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）