Biosensors-Basel
·
2022
·Vol. 13
·期 1
·22
·
IF 5.743
Lingyuan Xu,A. M. Abd El-Aty,Jing Zhao,Xingmei Lei,Xiuyuan Zhang,Yun Zhao,Xueyan Cui,Yongxin She,Fen Jin,Jing Wang,Maojun Jin,Bruce D. Hammock
In this study, a previously unreported 3-((4-(isopropylamino)-6-(methylthio)-1,3,5-triazin-2-yl) amino) butyric acid hapten was designed and synthesized. This maximized the exposure of the antigen-determinant isopropyl of prometryn to the immune system in animals to induce the production of anticipated highly specific anti-prometryn antibodies. The hapten has a molecular weight of 285.37 Da. The compound was confirmed by nuclear magnetic resonance hydrogen spectroscopy (1H NMR), nuclear magnetic resonance carbon spectroscopy (13C NMR), and high-resolution mass spectrometry (HRMS). By using the active ester approach, immunogens and coated antigens were created. Bovine serum albumin (BSA) was used as an immunogen, along with the successfully produced hapten, to immunize mice. The IC50value of mouse monoclonal anti-prometryn antibody (mAb) 7D4 (the quantity of analyte that generated 50% prometryn inhibition) was 3.9 ng/mL. The anti-prometryn mAb was of the IgG1 subclass. The IC20(80% binding level (B/B0) of prometryn)-IC80(20% binding level (B/B0) of prometryn) range of the anti-prometryn monoclonal antibody standard curve working range was 0.9–18.1 ng/mL. The prepared mAb has good characteristics because it can specifically recognize prometryn, and the cross-reaction rates for ametryn, desmetryn, and terbumeton were 34.77%, 18.09%, and 7.64%, respectively. The cross-reaction rate with the other seven triazines was less than 1%. The hapten structure proposed can serve as an additional tool for modulating selectivity in detecting triazines.Keywords:prometryn;hapten;monoclonal antibody;immunoassays
研究领域:
环境科学
·
生物化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2014
·
IF 3.107
Lin Luo,Zhen-Lin Xu,Jin-Yi Yang,Zhi-Li Xiao,Yong-Jun Li,Ross C. Beier,Yuan-Ming Sun,Hong-Tao Lei,Hong Wang,Yu-Dong Shen
Novel haptens were designed and synthesized to prepare antibodies against free histamine, but none resulted in producing suitable antibodies for developing an enzyme-linked immunosorbent assay (ELISA). However, an antiserum was obtained having high specificity and affinity to p-nitrobenzoylated histamine (NPHA), which can be easily formed from reaction between histamine and p-nitrobenzoic acid N-hydroxysuccinimide ester (PNBA-OSu) under mild conditions. Based on rabbit polyclonal antibodies, a competitive indirect ELISA (ciELISA) for histamine determination in foods was developed. After ciELISA and derivatization optimization, the assay showed good sensitivity, with limits of detection of 1.8 mg/kg, 93.6 μg/L, and 93.6 μg/kg in fish, red wine, and yoghurt, respectively, with negligible cross-reactivity with related biogenic amines and amino acids. Average recovery of histamine in fortified food samples ranged from 80.9% to 110.1% with coefficients of variation below 16.3%. Good correlation between the ciELISA and liquid chromatography–tandem mass spectrometry was obtained for spiked food samples.
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
FOOD AND AGRICULTURAL IMMUNOLOGY
·
2019
·
IF 2.398
Xiujin Chen,Zhaozhou Li,Fengxia Sun,Xiaotian Cao,Yao Wang,Li Cao,Hongli Gao,Dong Gao,Yawen Wang
Tebuconazole is widely applied for controlling pathogenic fungi of crops and proved hazardous to human being. To better measure tebuconazole residues, an enzyme-linked immunosorbent assay for screening tebuconazole residues in aqueous samples was established using an innovative hapten (hapten1) designed via molecular model. The work concentration of calibration standard curves based on aqueous matrices ranged from 0.51 to 10.71 ng mL–1. The cross-reactivities of other analogs were less than 6%. The detection limits of the assay were in the range of 0.197–0.297 ng mL–1 for tebuconazole in water samples. The average recoveries of tebuconazole in the spiked water samples varied from 76.76% to 107.52% with the variation coefficient scope of 4.54–13.26%. The obtained immunoassay was verified by gas chromatography coupled to mass spectrometry with high correlation coefficients (r > 0.9868). Herein, the proposed immunoassay offers a cost-effective, convenient, reliable tool for the determination of tebuconazole residues in aqueous samples.
研究领域:
环境科学
·
分析化学
·
毒理学
✓ 已验证
产品关联
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
Analytical Methods
·
2014
·Vol. 6
·期 12
·4228-4233
·
IF 1.938
Liqiang Liu,Huang Kuang,Chifang Peng,Libing Wang,Chuanlai Xu
Salbutamol (SAL) is a short-acting β2-adrenergic receptor agonist (β-agonist) forbidden in livestock production. To develop a more sensitive and specific detection method for SAL, a new hapten was synthesized, keeping the structure of benzene ring side of SAL while changing the tert-butyl group structure that SAL shares with other β-agonists. Ten mice were immunized with a SAL hapten–keyhole limpet hemocyanin (KLH) conjugate, and cells were selected within a urine sample at the cell fusion stage. The best monoclonal antibody (Mab) had an IC50 value of 0.31 ng mL−1 in PBST buffer and an IC50 value of 0.19 ng mL−1 in a swine urine sample diluted five times (with a lower B0 value), and had no cross-reactivity with clenbuterol and other β-agonists. These results show that the hapten design was successful and that the Mab can potentially be used in highly sensitive immunoassays for specifically monitoring the illegal use of SAL in real samples.
研究领域:
免疫学
·
药物检测
·
动物医学
✓ 已验证
产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Hongfei Yang,Hao Wen,Yu Si,Mingyue Ding,Yiting Liu,Ziyan Yu,Linwei Zhang,Jiacan Wang,Xiaoming Pan,Shiyun Han,Yiting Wang,Shixiang Wu,Jixiang Liang,Jiaxu Xiao,Rui Fang,Dapeng Peng
Altrenogest is one of the most commonly used steroid hormones; however, there are currently no relevant reports on monospecific molecular recognition elements and immunoassay methods for altrenogest. Herein, a computer-aided precise hapten design strategy was proposed for monospecific monoclonal antibodies (mAb) preparation. Based on this strategy, a monospecific and sensitive mAb-D7 was prepared for the first time. The mAb-D7 has the 50 % inhibitory concentration (IC 50 ) of 0.12 ng/mL for altrenogest and does not cross-react with other common steroid hormones. Additionally, a single-chain variable fragment (scFv) for altrenogest was constructed for the first time, which exhibits an IC 50 of 1.7 ng/mL for altrenogest. The molecular recognition mechanism studies showed the monospecific mAb-D7 to altrenogest originated from the amino acids PHE-94 and LEU-237, demonstrating the reliability of this strategy. Finally, two monospecific, rapid, and sensitive immunoassays were established for altrenogest in pork and pork liver for the first time.
研究领域:
生物化学
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免疫学
·
兽医学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD SCIENCE
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2022
·Vol. 87
·期 3
·1342-1354
·
IF 3.693
Jin-Yi Yang,Mei-Chan Xie,Xue-Cai Tan,Yuan-Xin Tian,Hong Wang,Zhen-Lin Xu,Ting-Ting Yuan,Yi-Mei Xiao,Yu-Dong Shen
The tadalafil-like compounds have appeared recently as adulterants in drinks and healthcare dietary supplements sourced from medicinal and edible food, which may cause illness and even death. In this work, the rationality of haptens was explored by computational chemistry and molecular simulation theories such as frontier molecular orbital (FMO)-based softness (S), three-dimensional (3D) structure, surface electrostatic potential (ESP), and lipophilic potential (LP). An antiserum from hapten H5 with the highest softness and maintaining the appropriate three-dimensional (3D) structure showed the optimal immunoassay performance, indicating an increasing softness was a critical factor for effective hapten. Based on the antibody induced by hapten H5, an indirect competitive enzyme-linked immunosorbent assay (icELISA) method for detecting multiple tadalafil-like adulterants was established. The icELISA showed a limit of detection (LOD), 50% inhibition concentration (IC 50 ), and a working range of 0.004–0.396, 0.89–4.27, and 0.094–16.71 ng/ml for tadalafil, amino tadalafil, acetamino tadalafil, nortadalafil, and N-desmethyl ent-tadalafil, respectively. The spiked recoveries of tadalafil-like adulterants in samples ranged from 84.9% to 116.2%. The results of the icELISA and HPLC-MS/MS methods had a good correlation for real samples with the R 2 of 0.9955. Specially, this work not only provided a convenient immunoassay method for measuring tadalafil-like adulterants in spirit drinks and dietary supplements in group-screening manner, but also suggested that softness was likely to be a general theory for rational hapten design. Practical Application Rapid monitoring of tadalafil-like adulterants in food samples is very necessary and important for consumers, regulatory agencies, and the food industry.
研究领域:
分析化学
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免疫学
·
药物化学
✓ 已验证
DOI
高影响因子
Food Chemistry-X
·
2024
·
IF 6.5
Kunying Nie,Jiali Zhang,Haitao Xu,Keyun Ren,Chunlei Yu,Qi Zhang,Falan Li,Qingqing Yang
Rapid simultaneous detection of multi-component adulteration markers can improve the accuracy of identification of gutter cooking oil in edible oil, which is made possible by broad-spectrum antibody (bs-mAb). This study used capsaicinoids (CPCs) and gingerol derivatives (GDs) as adulteration markers, and two broad-spectrum haptens (bs-haptens) were designed and synthesized based on a reverse design strategy of molecular docking. Electrostatic potential (ESP) and monoclonal antibodies (mAbs) preparation verified the strategy's feasibility. To further investigate the recognition mechanism, five other reported antigens and mAbs were also used. Finally, the optimal combination (Hapten 5-OVA/1-F12) and key functional groups (f-groups) were determined. The half maximal inhibitory concentration (IC 50 ) for CPCs-GDs was between 88.13 and 499.16 ng/mL. Meanwhile, a preliminary lateral flow immunoassay (LFIA) study made practical monitoring possible. The study provided a theoretical basis for the virtual screening of bs-haptens and simultaneous immunoassay of multiple exogenous markers to monitor gutter oil rapidly and accurately.
研究领域:
食品科学
·
免疫学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2024
·
IF 4.8
Ghulam Mujtaba Mari,Weilin Wu,M. Farooque Hassan,Ambreen Laghari,Hidayatullah Soomro,Aisha Talpur,Zhanhui Wang,Xuezhi Yu
Nitrovin, classified as a nitrofuran drug, has been banned in numerous countries due to its carcinogenic effect on human health. However, existing analytical techniques for its detection are laborious, requiring an overnight derivatization process lasting 16 h. In this study, novel haptens were designed to prepare antibodies against aminogunidine (AGD), the metabolite of nitrovin, and then developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) method to detect AGD from fish samples without derivatization for the first time. Computer-assisted molecular modeling predicted the efficiency of novel haptens by conformational studies and electronic analysis. The results were consistent with the predicted results in producing high-affinity monoclonal antibody (mAb) against AGD. The half-inhibition concentration (IC 50 ) value of icELISA was 0.80 ng mL −1 for AGD, with a limit of detection of 0.27 μg kg −1 and recoveries of 89.8%–97.2%. The results showed that this new hapten design and synthesis provided a great prospect to obtain high-affinity mAb for AGD. This developed icELISA substantially avoided the conventional and lengthy derivatization process and was an effective method for the direct detection of AGD in fish tissue without derivatization.
研究领域:
药理学
·
毒理学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2025
·
IF 9.8
Feng He,Lingling Guo,Liqiang Liu,Xinxin Xu,Chuanlai Xu,Hua Kuang,Maozhong Sun
Maleic hydrazide (MH), a selective herbicide and plant growth inhibitor, poses risks to ecological balance and human health due to its widespread use. In this study, we developed a simple, reliable, and rapid detection method for MH. Four haptens were analyzed using computational chemistry simulations, and a matrix effect-enhanced screening method was used to obtain the anti-MH monoclonal antibody, with a half-maximal inhibitory concentration (IC 50 ) of 0.157 μg/mL. An immunochromatographic assay was developed for detecting MH in lake water, soil, onion, garlic, potato, and tobacco samples. The IC 50 values ranged from 0.590 to 9.419 mg/kg, with a detection limit of 0.047–1.614 mg/kg. Spiked recovery and real sample analyses demonstrated results consistent with instrumental methods, achieving recoveries of 89.2 %–111.3 % and CV < 7 %. These findings confirm the method's suitability for detecting MH in food and environmental samples, fulfilling the requirements for on-site rapid detection in resource-limited settings.
研究领域:
分析化学
·
计算化学
·
免疫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Liang Luo,Yingjie Zhang,Xiangjun Zhao,Weilin Wu,Jie Fei,Xuezhi Yu,Kai Wen,Jianzhong Shen,Yantong Pan,Zhanhui Wang
Xylazine (XYL) is an illicit adulterant in opioids and an approved veterinary sedative drug, which has been abused, misused, and residued in food samples, endangering people health, causing drug-facilitated crimes and even death. Immunoassay used antibody as core biomaterial could to achieve highly sensitive and rapid detection screening purpose for XYL in situ . Here, we rationally designed four novel XYL haptens with different spacer arms to produce antibodies with high affinity and specificity. Ten monoclonal antibodies (mAbs) were obtained and mAb 7H5 showed a high affinity with IC 50 of 0.23 ng mL −1 and ignorable cross-reactivity for the other eight analogs. One highly sensitive indirect competitive ELISA (icELISA) and lateral flow immunoassay (LFIA) were established based on heterologous haptens for XYL detection in a series of human urine and food samples. The limit of detections (LODs) of the icELISA were 0.10–2.24 μg L −1 with recovery of 83.5 % to 128.6 % and CV below 15.8 % in different samples, while, the visual limits of detection (vLOD) of LFIA were 0.10–1.80 μg L −1 with the cut-off value of 0.60–4.80 μg L −1 . In addition, the molecular recognition mechanism of mAbs was explored. The study provides powerful tools for rapid screening of XYL in human urine and food samples for the first time.
研究领域:
免疫学
·
药物检测
·
食品安全
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Zizhe Li,Qianqian Cui,Qingyue Li,Changwei Luo,Mengtian Chen,Beibei Feng,Huan Li,Tong Bu,Yexuan Mao,Meng Dang,Xianqing Huang,Lianjun Song,Dapeng Peng,Xiya Zhang
To overcome limitations with respect to the low sensitivity of rapid immunoassay methods due to dilution of the extract in extraction solution, ultra-sensitive antibodies are needed. The linker-tethering site between the target and carrier protein , the length of the spacer arm , and the atomic charge of the linker-tethering site between the target and spacer arm in haptens design play a pivotal role in the preparation of ultra-sensitive antibodies. In this study, carbendazim (CBZ), a systemic broad-spectrum fungicidal pesticide, was used as model compound. 2-Aminobenzimidazole was chosen as the common skeletal structure and reacted with different groups, such as alkanes and urine groups, to form haptens H1 (previously reported), H2 (novel), and H3 (novel), respectively. Subsequently, eight monoclonal antibodies (mAbs) were generated using these three haptens , which exhibited notable variation in sensitivity. The half maximal inhibitory concentration (IC 50 ) value of mAb 4B11 based on hapten H3 against CBZ was 0.04 ng/mL, which was seven times better than that of mAb 5B10 (IC 50 value of 0.28 ng/mL) prepared using hapten H2 and 129 times better than that of mAb 6D5 (IC 50 value of 5.15 ng/mL) prepared using hapten H1. Finally, the optimal antigen–antibody combination was employed to establish a sensitive colloidal gold lateral flow immunoassay (CG-LFA) for CBZ detection in vegetables and fruits with convenient sample pretreatment . The developed CG-LFA shows promising practical utility and prospects for application due to its low limit of detection (0.10–0.18 ng/mL) and high recovery ratios (76.0%–96.1%) in six different samples.
研究领域:
农业科学
·
免疫学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）