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
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 9
·1-13
·
IF 5.3
Kang Jia,Qi Meng,Wang Yasen,Li Linchun,Zhou Xinda,An Xiaoshuo,Hammock Bruce D.,Zhang Jinlin,Huo Jingqian
Nanobodies specifically recognizing atrazine were successfully obtained through immunization of alpacas and phage display libraries, and an indirect competitive immunoassay (ic-ELISA) based on nanobodies was constructed with an IC 50 of 0.062 μg/mL and a minimum detection limit of 0.01 μg/mL. An Au@PtNPs with excellent colorimetric performance, high catalytic activity, and high stability was synthesized. The combination of Au@PtNPs and nanobodies led to the development of a direct competitive immunoassay (dc-ELISA) with higher specificity, convenience, lower cost and an IC 50 value of 0.032 μg/mL which was lower than the ic-ELISA. Based on the Au@PtNPs and nanobodies, a novel LFIA model for qualitative and semi-quantitative detection of atrazine was constructed with a visual detection limit of 5 ng/mL. The present study is of great significance for the construction of atrazine immunoassay combining nanobodies and nanoenzymes, which provides a sensitive, specific, simple and reliable method for the detection of atrazine in the environment.
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研究领域:
分析化学
·
免疫学
·
纳米科技
·
环境科学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2024
·
IF 1.7
Jiaxin Wang,Wenhua Shang,Shibei Shao,Jialin Zhang,Qiuyang Wu,Min He,Licai Ma,Jianzhong Shen,Zhanhui Wang,Kai Wen
Paxilline (PAX) is a tremorgenic mycotoxin that is thought to be associated with various syndromes in animals. To facilitate the determination of this toxin, a hapten was chemically synthesized and then respectively conjugated to Bovine Thyroglobulin (BTG) and Bovine serum albumin (BSA) for antibody production. Monoclonal antibody (mAb) 4F8 was finally obtained and used to establish an accurate and sensitive indirect competitive ELISA (icELISA). Various parameters that could impact the performance of icELISA were optimized, including the ion concentration, pH value and organic solvent. Under optimal condition, this assay exhibits an IC50 value as low as 0.84 µg L−1. The limit of detection (LOD) of the established icELISA method for PAX was 0.03–0.3 µg L−1. In conclusion, this study provided a rapid, sensitive, and high-throughput immunoassay for the detection of PAX in grains.
研究领域:
免疫学
·
食品科学
·
毒理学
✓ 已验证
产品关联
DOI
高影响因子
CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
·
2018
·
IF 0.824
Ran LI,Ze-Jia LIN,Jin-Yi YANG,Zhen-Lin XU,Hong WANG,Hong-Tao LEI,Yuan-Ming SUN,Yu-Dong SHEN
A high-affinity polyclonal antibody was prepared by immunizing animals with haptens FFD and FFM. Under the optimal combination of coating antigen and antibody, an indirect competitive enzyme-linked immunosorbent assay (icELISA) for simultaneous detection of florfenicol and thiamphenicol residues in animal meat and urine samples was developed. The icELISA showed an IC 50 value of 1.32 ng mL −1 for florfenicol and 2.13 ng mL −1 for thiamphenicol, respectively. The linear ranges were from 0.31 to 5.61 ng mL −1 with a limit of detection of 0.12 ng mL −1 for florfenicol, and 0.41 to 11.2 ng mL −1 with a limit of detection of 0.15 ng mL −1 for thiamphenicol, respectively. The average recoveries of florfenicol and thiamphenicol in spiked samples ranged from 77.2% to 116.0% with a relative standard deviation of less than 15%. Therefore, this proposed icELISA provided a valid detection method for florfenicol and thiamphenicol residues in animal tissue and urine samples.
研究领域:
免疫学
·
食品安全检测
✓ 已验证
DOI
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Yaqi Jin,Zihui Jin,Di Sun,Yifan Liu,Bo Xu,Qiyang Zhao,Yue He,Lei Gong,Jing Li,Yaohai Zhang,Yongliang Cui
Spiropidion developed by Syngenta shows high insecticidal and acaricidal activity against a wide range of sucking pests. In this study, according to the structure of spiropidion, two haptens were synthesized by introducing carboxyl groups from the ester group. After cell fusion, a monoclonal antibody (mAb 8B5) of spiropidion was obtained. The IC 50 of the established heterologous indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was 7.36 ng/mL, and its working range was 1.75–34.92 ng/mL. The average recoveries were 76.05–124.78% in the Yangtze River and citrus samples. Moreover, the ic-ELISA results of 15 citrus samples agreed well with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Overall, the established ic-ELISA could be applied for the spiropidion residue monitor in food and agricultural samples.
研究领域:
农业科学
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2020
·
IF 6.306
Yi-Fang Li,Zheng-Zhong Lin,Cheng-Yi Hong,Zhi-Yong Huang
Due to complex matrixes and specific reagent deficiency, the rapid detection of histamine is still a challenge to date. Based on the high peroxidase-like activity of iron-cobalt co-doped carbon dots, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established for histamine detection using the mimic enzyme labeled with histamine antibody (His-Ab). Through the competitive binding of the labeled His-Ab to solid-phase and sample antigens, histamine content was detected with a linear range of 2.5–150 μg mL −1 . The detection limit based on 3σ/K was 0.50 mg kg −1 , which was much lower than those of commercial His-kit and HPLC methods. The ic-ELISA method was applied to histamine detection in fish samples with the recovery of (103.4 ± 0.5)%, which was in accord with those of commercial His-kit and HPLC methods. The results indicated that the established ic-ELISA method was suitable for rapid detection of histamine in fish samples with high accuracy, sensitivity and stability.
研究领域:
食品科学
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Mengxiang Wang,Jinxing Song,Yanze Li,Junru Sun,Lei Zhou,Zhuoya Sun,Angke Zhang,Guoqing Zhuang,Pingan Xia,Gaiping Zhang,Yanan Wu
African swine fever has caused huge losses to the global pig industry. In the absence of effective vaccines, reliable detection methods are crucial. The p30 protein of ASFV is often used as a target for detection due to its high antigenicity in the early stage of virus replication. In this study, we modified the p30 protein using magnetic nanoparticles to increase its specific surface area and suspension in solution. Based on this modified p30 protein, we optimized the experimental steps to determine the best reaction conditions and then established a double probe competitive ELISA method. The established detection method can produce results within 45 min, with repeatability between and within batches <15 %, and it does not react with other pathogens, which demonstrates high specificity. In conclusion, we modified the ASFV p30 protein with magnetic nanoparticles and established a rapid, ultra-sensitive, and convenient detection method for ASF based on the modified p30. This also provides a new reference for the rapid detection of other veterinary diseases.
研究领域:
兽医学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2013
·Vol. 138
·期 17
·5151-5156
·
IF 3.969
Zhiwei Qie,Baoan Ning,Ming Liu,Jialei Bai,Yuan Peng,Nan Song,Zhiqiang Lv,Ying Wang,Siming Sun,Xuan Su,Yihong Zhang,Zhixian Gao
Fast detection is important in screening large-scale samples. This study establishes a direct competitive ELISA method (dcTELISA) based on an enzyme thermistor for fast atrazine (ATZ) detection. ATZ competes with β-lactamase-labeled ATZ (ATZ–E) for the binding sites on anti-ATZ monoclonal antibody (mAb). The mAb are covalently bound to Controlled Pore Glass (CPG) in an immunoreactor to form immunocomplexes with ATZ and ATZ–E. Several parameters of biosensor performance were optimized, such as the ATZ–E concentration, concentration and nature of the substrate, flow rate, and effect of temperature on the sensor response. After optimization, the assay time for a single sample was 12 min. The work process and result were compared with those of high-performance liquid chromatography (HPLC). The detection results exhibited a recovery rate of 88% to 107% in ATZ-spiked fresh cut corn stalks and silage samples. The results obtained via dcTELISA had good correlation with that of HPLC, and the biosensor response was reproducible and stable even when used continuously for over 4 months. All these properties suggested that the fast detection method, dcTELISA, may be used to detect pesticide residue in large-scale samples.
研究领域:
环境科学
·
生物技术
✓ 已验证
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
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 104
·期 17
·7345-7354
·
IF 3.53
Liu Yuan,Liu Dan,Shen Chen,Dong Sa,Hu Xiaodan,Lin Manman,Zhang Xiao,Xu Chongxin,Zhong Jianfeng,Xie Yajing,Zhang Cunzheng,Wang Donglan,Liu Xianjin
Abstract Pyrethroids are insecticides that are widely used in rural and urban areas worldwide. After entering the environment, pyrethroids are rapidly metabolized or degraded by various biological or abiotic methods. In this study, a single-chain variable fragment (scFv) which could simultaneously detect three pyrethroid metabolites was constructed based on a hybridoma raised against 3-phenoxybenzoic acid (3-PBA). By molecular docking, it showed that there were hydrogen bonds, hydrophobic interactions, CH-π interaction, and cation-π interaction between 3-PBA and its scFv. All the contact residues contributing to hydrogen bonds are located in VH-CDR2 or its neighboring region, and two of them were mutants of the closest germline sequence. Based on competitive ELISA, the half maximal inhibitory concentration (IC 50 ) of the scFv for 3-PBA, 3-phenoxybenzaldehyde (PBAld), and 3-phenoxybenzyl alcohol (PBAlc) were calculated to be 0.55, 0.59, and 0.63 μgmL −1 , respectively. The scFv also showed 23.91%, 13.41%, 1.15%, 1.00%, and 0.56% cross-reactivity with phenothrin, deltamethrin, fenvalerate, beta-cypermethrin, and fenpropathrin. The broad specificity of the scFv may be due to its hapten design. The scFv could be employed in class-specific immunoassays for pyrethroid metabolites with phenoxybenzyl (PB) group. It is also potentially used for characterizing degradation of pyrethroids or detecting PBAlc (PBAld) alone, and the detection results should be confirmed by other selective methods. Key points • A scFv which can simultaneously detect 3-PBA, PBAlc, and PBAld was constructed. • Antibody informatics and binding mode of the scFv were obtained. • The reason for its broad specificity was discussed. • It could be used to monitor single or multi-pyrethroid metabolites with PB group.
研究领域:
环境科学
·
免疫学
✓ 已验证
DOI
PubMed
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2017
·Vol. 24
·期 8
·7042-7050
·
IF 2.741
Zhang Dai,Zhou Lifang,Lei Yajing,Zhou Zhan,Zhou Jie,Chen Shuqing
Diethylstilbestrol (DES) is used as a kind of animal feed additive and affects people’s health through the food chain. The purpose of this study is to detect the residue level of DES in 576 human urine samples directly. DES-BSA was used to immunize Balb/c mice. The monoclonal antibody was produced by hybridoma that was screened through cell fusion techniques. Finally, we developed the indirect competitive ELISA method to analyze 576 human urine samples from Zhejiang Province, China. The IC 50 of this method was 3.33 ng/mL. The LOD and LOQ were 0.16 and 0.54 ng/mL. Linear range of the standard curve was from LOD to 12.50 ng/mL. There was no cross-reactivity with two kinds of estrogens and two structural analogs with DES. Five hundred seventy-six urine samples were analyzed by the indirect competitive ELISA method, and the detection rate was 98.78%. The mean concentration and geometric mean were 4.70 and 3.50 ng/mL. The indirect competitive ELISA method based on monoclonal antibody was sensitive and reliable for the detection of DES in human urine samples. The results warned us to pay more attention to human health and food safety.
研究领域:
分析化学
·
毒理学
·
公共卫生
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Lakshani Madushika,Yongming Huang,Menghan Sun,Yuxuan Liu,Linfang Lu,Xiaoli Li,Na Li,Kang Jiang,Sumei Ling,Shihua Wang
Moniliformin (MON) is a toxic secondary metabolite from Fusarium species. The natural contamination of MON in cereals and cereal by-products, poses a risk of exposure to MON. However, so far, no immunoassay method has been reported to detect MON in field samples. In this study, anti-MON monoclonal antibody (mAb) with high affinity (1 × 10 8 ) and isotype of IgG1 was subjected to establish indirect competitive ELISA (ic-ELISA) and immunochromatographic test strips (ICTS). The optimized ic-ELISA resulted in the lowest detection limit (LOD) of 1.55 μg/mL. The LOD value for gold nanoparticles (AuNP) -based test strip was 0.83 μg/mL. The multi branched gold nano-flower particles (AuNFs) reported higher sensitivity with LOD of 0.38 μg/mL. Moreover, the developed test strips exhibited a high specificity without cross-reactivity to related competitive toxins. In conclusion, immunoassay methods were successfully developed to detect MON in field samples with a short detection time to ensure food security.
研究领域:
食品科学
·
免疫学
·
纳米材料科学
✓ 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）