Food Bioscience
·
2023
·
IF 5.2
Jialin Hu,Xinxin Xu,Liguang Xu,Hua Kuang,Chuanlai Xu,Lingling Guo
Wheat gliadin is one of the most important causes of gluten sensitivity. This study aimed to develop a sandwich enzyme-linked immunosorbent assay (ELISA) and gold nanoparticle-based lateral flow (LFIA) strips to detect wheat allergen in food. Through cell screening, ten monoclonal antibodies (mAbs) against gliadin and after optimization were obtained, a pair of mAbs suitable for its detection (capture antibody: mAb 7; detection antibody : HRP-labeled mAb 6) were identified. Based on two of these antibodies, a sandwich ELISA with limit of detection (LOD) of 60 ng/mL in negative milk, and more importantly, negligible cross-reactivity to other allergens was developed. The average recoveries for gliadin in negative milk were 99.16%–100.07% using the sandwich ELISA. We also developed LFIA strips for the rapid detection of gliadin with visual limit of detection (vLOD) of 25 ng/mL and calculated LOD value of 6.56 ng/mL in negative milk. The test results of positive samples obtained from the LFIA strips were highly consistent with those of the sandwich ELISA. Thus, the developed LFIA strip is an effective and reliable tool for the rapid and on-site detection of wheat allergen in milk.
研究领域:
食品科学
·
免疫学
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DOI
高影响因子
Food Bioscience
·
2024
·
IF 4.8
Jiao Li,Panpan Cui,Zhijia Wang,Yang Lu,Yuan Ding,Xiude Hua
Haloxyfop-P-methyl, a widely used aryloxyphenoxypropionate herbicide, has been identified as a potential hazard to ecological security and human health. Consequently, it is essential to develop superior methods for detecting haloxyfop-P-methyl. In this study, 12 peptidomimetics and 4 anti-immunocomplex peptides were isolated from phage-displayed peptide libraries, which can specifically recognize the anti-haloxyfop-P-methyl antibody and the immunocomplex formed by haloxyfop-P-methyl and its antibody, respectively. Following the sensitivity assessment of each phage-displayed peptide, competitive and noncompetitive phage enzyme-linked immunosorbent assays (P-ELISAs) were developed using the best phage-displayed peptides. The half inhibition concentration (IC 50 ) and half saturation concentration (SC 50 ) of competitive and noncompetitive P-ELISAs were determined to be 8.15 and 1.83 ng/mL, respectively, representing the 5-fold and 22-fold increase in sensitivity compared to previously reported immunoassay. The competitive P-ELISA exhibited 2.2%–114.8% cross-reactivities (CRs) to haloxyfop-methyl, haloxyfop and quizalofop-p-ethyl, while the noncompetitive P-ELISA showed no significant CR (<0.1%) except for haloxyfop-methyl (CR = 41.6%). The average recoveries of two P-ELISAs in soil, soybean, potato, and cabbage samples ranged from 78.4% to 104.3% and 76.8%–95.2%, respectively. Additionally, the results of P-ELISAs for blind samples exhibited a good correlation with those obtained from ultra-performance liquid chromatography/tandem mass spectrometry.
研究领域:
环境科学
·
分析化学
·
毒理学
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DOI
高影响因子
ANALYTICAL SCIENCES
·
2019
·Vol. 35
·期 2
·133-140
·
IF 1.618
Longjie ZHONG,Jiadi SUN,Ying GAN,Shuqi ZHOU,Zijian WAN,Quchao ZOU,Kaiqi SU,Ping WANG
Saxitoxin (STX) is one of the paralytic shellfish poisons (PSP) that endanger people’s health. It is necessary to develop methods for the on-site rapid detection for STX in order to prevent safety accidents. An enzyme-linked immunosorbent assay (ELISA) is timesaving and effective, but it is not suitable for large-scale in-field tests due to the expensiveness of commercial ELISA kits and the bulkiness of a microtiter plate reader (MTPR). In this study, a portable smartphone-based colorimetric analyzer (SBCA) with a cost-effictive enhanced gold nanoparticle-based ELISA (EGNB-ELISA) was proposed for STX detection. In a bicinchoninic acid (BCA) protein assay (R2 = 0.9939) and a glucose assay (R2 = 0.9937), SBCA was shown to be in good agreement with MTPR. EGNB-ELISA had a 12.5-fold lower detection limit (0.4 ng/mL) and a lower detection range (1 – 50 ng/mL, Y = 0.4037X + 0.3564, R2 = 0.9797) than the classical ELISA. The recovery rate ranged over 89.1 – 112.2%. The whole detection system, combining both homemade SBCA and ENGB-ELISA, is expected to satisfy the needs of on-site STX sample tests to guarantee seafood safety.
研究领域:
食品安全
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分析化学
·
纳米技术
·
环境科学
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产品关联
DOI
PubMed
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
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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
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DOI
PubMed
高影响因子
Drug Testing and Analysis
·
2014
·Vol. 6
·期 10
·1020-1027
·
IF 2.84
Yajing Lei,Qian Zhang,Lizheng Fang,Muhammad Sajid Hamid Akash,Kanwal Rehman,Zhiming Liu,Weixing Shi,Shuqing Chen
We simultaneously set up two competitive (direct and indirect) enzyme-linked immunosorbent assays (ELISAs) based on the same antibody for estimation of cotinine (COT) in pregnant women especially and population generally exposed to environmental tobacco smoke. The results show that the limits of detection (LODs) for direct competitive ELISA and indirect competitive ELISA were 0.04 μgL -1 and 0.1 μgL -1 , respectively. Direct competitive ELISA was found to be more sensitive than indirect competitive ELISA. Thereafter, we applied our direct competitive ELISA for the detection of COT from urinary samples taken from 450 volunteers from the Zhejiang Province of China. COT was detected in 100% of participants with concentration ranging from LOD to 5358.0 μgL -1 . The GM and 95th percentile concentration of COT in pregnant women were 6.3 μgL -1 and 57.2 μgL -1 , respectively. Males had statistically higher COT concentrations than females ( P < 0.0001), active smokers had statistically higher COT concentrations than non-smokers ( P < 0.0001), whereas, non-pregnant women were found to have higher COT concentration than pregnant women. We conclude that our developed direct competitive ELISA is useful for detecting the COT in urinary concentration of human. The human urinary data obtained in this study indicated that common people generally and pregnant women especially were highly exposed to COT. Further studies are needed to focus on the sources of exposure, potential health effects and risk assessment of exposure to COT. Copyright © 2014 John Wiley & Sons, Ltd.
研究领域:
分析化学
·
环境卫生
·
公共卫生
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DOI
PubMed
高影响因子
TALANTA
·
2026
·
IF 6.1
Tianyu Lan,Jiexin Zheng,Bo Xu,Kerui Dan,Chunlei Cao,Feng Sun,Qiyang Zhao,Yue He,Jing Li,Yongliang Cui,Yaohai Zhang
Cyenopyrafen is an effective pyrazole acaricide used for mite control on various crops. This study generated high-affinity monoclonal antibodies specific to cyenopyrafen and developed a high-throughput indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) validated for sensitivity, specificity, and reproducibility. The ic-ELISA demonstrated an IC 50 of 32.75 ng/mL and a dynamic range of 2.33-293.19 ng/mL, with negligible cross-reactivity (<0.1%) with related acaricides. Recovery rates in citrus samples were 88.18%–105.89%. A strong correlation (R 2 > 0.95) was detected between the results of ic-ELISA and UPLC-MS/MS, highlighting the assay’s reliability. This immunoassay offers a reliable approach for the determination of cyenopyrafen in the tested citrus and apple samples, and may serve as a basis for broader applications with potential utility in regulatory frameworks.
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DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2021
·
IF 3.365
Yuanyuan Ding,Xiaoqian Li,Qingpeng Gao,Xinyan Dong,Liyun Kong,Shengli Han,Tao Zhang,Langchong He
Mas-related G-protein-coupled receptor X2 (MRGPRX2) has recently been reported to be associated with anaphylaxis. Detection of MRGPRX2 levels in human peripheral blood might serve as a powerful tool for predicting the predisposition of patients to anaphylactic reactions. For rapid measurement of MRGPRX2, we established a paper-based double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antibody and horseradish peroxidase (HRP)-labelled rabbit polyclonal antibody as capture antibody and detection antibody, respectively. We avoided chemical functionalization of the cellulose paper by introducing bovine serum albumin (BSA) to provide COOH and NH 2 groups for covalent immobilization of the capture antibody. Through amide condensation , a two-layer immobilization strategy was applied with BSA-BSA and BSA-capture antibody networks as the first and second layers, respectively. This strategy improved the quantity, activity and stability of the immobilized antibody . We then established a paper-based ELISA to detect MRGPRX2 in human peripheral blood. Our method is less laborious, easier to implement, and more cost-effective than conventional ELISA, while offering similar sensitivity, specificity, and accuracy. Therefore, it could serve as an innovative clinical point-of-care diagnostic tool, especially in areas that lack advanced clinical equipment.
研究领域:
医学
·
免疫学
·
生物化学
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DOI
高影响因子
TALANTA
·
2025
·
IF 6.1
Haiyun Deng,Weipeng Yu,Zhuqiao Fan,Jinfang Xue,Yanhua Gu,Xiaoqing Tang,Xiaomian Zhou,Nan Yu
Enzyme-linked immunosorbent assay (ELISA) is widely employed. Enhancing the surface of ELISA plate wells to increase the number of immobilized antibody molecules per unit area represents a viable strategy for improving the assay's detection sensitivity. Chitosan is particularly suitable for biomolecule attachment due to its porous structure and high density of amine groups. We aimed to enhance ELISA performance by applying a chitosan hydrogel layer onto polystyrene (PS) surfaces. The primary challenge was the instability of the hydrogel membrane formed on the PS surface, which was susceptible to detachment and cracking. To address this, a standardized three-step protocol was developed for the immobilization of chitosan hydrogel onto PS wells, comprising O 2 plasma pretreatment, vacuum drying for 3 h to enhance membrane adhesion, and glutaraldehyde-based chemical activation. The modified PS microplates exhibited stability for at least 1 year. The chitosan hydrogel membrane-based ELISA for alpha-fetoprotein (AFP) detection (AFP-CHM-ELISA) demonstrated excellent linearity (R 2 = 0.9992), with a limit of detection (LOD) of 1.98 ng/mL—comparable to the 1.00 ng/mL LOD of chemiluminescence—achieving a sensitivity approximately 2.5 times higher than that of conventional ELISA (5 ng/mL), and an extended upper linear range of 280 ng/mL. Intra- and inter-batch coefficients of variation were below 6 %, with spiked serum recovery rates ranging from 99.67 % to 103.48 %. Comparative analysis revealed a 95.54 % agreement with commercial chemiluminescence methods, along with a strong correlation (R 2 = 0.9236). This chitosan hydrogel-based surface modification approach significantly improves ELISA performance and demonstrates promising potential for diagnostic applications.
研究领域:
免疫学
·
肿瘤学
·
分子诊断
·
生物材料
·
临床检验
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2017
·
IF 5.401
Yi Liang,Xiaolin Huang,Xirui Chen,Wenjing Zhang,Guo Ping,Yonghua Xiong
A novel direct competitive plasmonic enzyme-linked immunosorbent assay (dc-pELISA) was applied to detect ochratoxin A (OTA) with naked eyes. In this assay, horseradish peroxidase (HRP) + hydrogen peroxide (H 2 O 2 ) + tyramine (TYR)-induced gold nanoparticle (AuNP) aggregation was considered as a signal output; AuNP aggregation could be triggered through the phenol polymerization of TYR , which was induced by hydroxyl radicals from HRP-catalyzed H 2 O 2 ; OTA-labeled catalase (CAT) was used as a competing antigen to consume H 2 O 2 . Owing to the combined advantages of ultrahigh CAT catalytic activity for H 2 O 2 and dual-color responses (red and blue) generated through AuNP aggregation, the proposed method was highly sensitive and thus could be employed with naked eyes to detect OTA qualitatively with a cut-off limit of 150 pg/mL. Our method also demonstrated a good dynamic linear range (12.5–150 pg/mL) for quantitative OTA determination with a reliable correlation coefficient of R 2 = 0.992, a half-maximal inhibitory concentration of 84.75 pg/mL, and a detection limit of 17.8 pg/mL. In brief, this newly-designed technique is considerably suitable for high-throughput screening detection or point-of-care diagnostics in resource-constrained regions because of the easy readout of results by naked eyes without the use of advanced detection equipment.
研究领域:
食品科学
·
分析化学
·
生物医学工程
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DOI
高影响因子
Analytical Methods
·
2026
·
IF 3
Ruoran Shi,Jingyao Liu,Honglin Ren,Yansong Li,Jiaqi Hou,Shaohui Hu,Yuzhu Wang,Chengwei Li,Bo Zhang,Ziqi Xing,Yuyang Ding,Fang Yang,Shiying Lu,Qiang Lu,Zengshan Liu,Xiaoxu Wang,Pan Hu
Listeria monocytogenes (LM) is one of the eight species within the Listeria genus and one of the most lethal foodborne pathogens. Accurate and rapid detection is crucial for preventing infections caused by this bacterium. Nanobodies, with the advantages of strong affinity, high specificity, low off-target effects, excellent thermal stability, and good solubility, have emerged as a novel reagent for pathogen detection. InlG (Internalin G), a surface-anchored virulence protein accessible on the bacterial surface, was selected as the target antigen to ensure high specificity and effective antibody binding. This study expressed and purified the inlG protein by prokaryotic expression. High-specificity nanobodies targeting the inlG protein were screened using phage display technology and were then used as capture antibodies to establish an indirect sandwich ELISA detection method. The established indirect sandwich ELISA assay demonstrates excellent reproducibility (intra-assay CV: 2.39% and inter-assay CV: 2.78%) and high specificity, with slight cross-reaction with Escherichia coli, and no significant cross-reaction with other food-borne pathogens. The limit of detection (LOD) was 1.0 × 105 CFU mL−1 and limit of quantitation (LOQ) was 3.0 × 105 CFU mL−1 (S/N = 10, CV ≤ 10%). Spiked recovery rates ranged from 96.0% to 107.5% in five food matrices (beef, fish, milk, soft cheese, and deli turkey), with an RSD ≤ 4.68%. Furthermore, these nanobodies targeting InlG may serve as penicillin for controlling L. monocytogenes infections, and chemical residues benzylpenicilloic acid (BPNLA) risk, addressing the challenge of antibiotic resistance.
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高影响因子
FOOD CONTROL
·
2019
·
IF 4.248
Yanbo Wang,Qinqin Qi,Jinru Zhou,Huan Li,Linglin Fu
Fish has been recognized as one of the “Big-Eight” allergenic food sources with parvalbumin (PV) being one of the major allergens. Here, we report a novel immunomagnetic beads-derived enzyme-linked immunosorbent assay (ELISA) method which employs antibody functionalized graphene oxide (GO) and gold nanoparticles (AuNPs) to amplify the detection signal of PV. The GO and AuNPs are modified with monoclonal antibody (mAb) and HRP-labeled secondary antibody respectively to detect the PV captured by amino-functionalized magnetic beads. With the amplification effect of the antibody modified GO and AuNPs, the proposed ELISA could selectively detect PV with a detection limit as low as 4.29 ng/mL when using pure PV. Moreover, the recovery rates are found to range from 89.82% to 115.37% in the spiked samples. The feasibility in real-world fish sample analysis further shows that the proposed immunomagnetic beads-derived ELISA is a highly promising approach for PV detection and pave a new way for food safety analysis.
研究领域:
食品科学
·
纳米技术
·
分析化学
·
免疫学
✓ 已验证
产品关联
DOI
高影响因子
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