International Journal of Environmental Research and Public Health
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2013
·Vol. 10
·期 7
·2897-2905
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IF 1.998
Juan Peng,Shanshan Song,Liguang Xu,Wei Ma,Liqiang Liu,Hua Kuang,Chuanlai Xu
We have established a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) based on two monoclonal antibodies (mAb) to measure the content of the major peanut allergen Ara h 1 in foods. Two mAbs were selected out of 12 murine hybridoma cells secreting Ara h 1-specific antibody. Using mAb 6 as the capture antibody and HRP-labelled mAb 4 as the detection antibody, the limit of detection (LOD) the assay was 0.34 ng/mL. Cross-reaction analysis showed that this method was strongly specific and had no cross-reactions with Ara h 2, pea protein or soy protein. Sample analysis showed that this ELISA was a useful tool to monitor peanut allergens in food products by measuring Ara h 1 content.Keywords:peanut allergen;Ara h 1;monoclonal antibody;sandwich ELISA
研究领域:
食品科学
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免疫学
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分析化学
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DOI
PubMed
高影响因子
Analytical Methods
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2014
·Vol. 6
·期 13
·4742-4745
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IF 1.938
Liqiang Liu,Dezhao Kong,Changrui Xing,Xun Zhang,Hua Kuang,Chuanlai Xu
Lactoferrin (LF) content in infant milk powder has been strictly regulated by many governments and there is a need for convenient and reliable assays. Using hybridoma techniques, fourteen monoclonal antibodies (mAbs) active against LF were prepared. Two antibodies (mAb2 and mAb3), recognizing spatially distant epitopes of LF, were selected to establish a sandwich enzyme-linked immunosorbent assay (ELISA). A solution of mAb3 (1 μg mL−1) was coated onto micro-titer plates for LF capture while mAb2 labeled with horseradish peroxidase (2.2 μg mL−1) was used as a detection antibody. Under optimized conditions, the proposed sandwich ELISA was evaluated, and linearly responded to LF standards in a range of 5–600 ng mL−1 and the limit of detection was 3.23 ng mL−1. Lactoferrin samples were able to be determined after simple dilution, and the recovery in fortified milk powder averaged between 98% and 109%. The developed assay showed both high specificity (with no obvious cross-reactivity with related proteins) and reproducibility (the coefficient of variation ranged from 4.5% to 7.1%), indicating the utility of this sandwich ELISA in LF monitoring.
研究领域:
免疫学
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食品科学
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分析化学
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DOI
高影响因子
FOOD AND AGRICULTURAL IMMUNOLOGY
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2012
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IF 0.714
Xiaofang Deng,Liqiang Liu,Wenwei Ma,Chuanlai Xu,Libing Wang,Hua Kuang
In this study, two anti-peanut agglutinin (PNA) polyclonal antibodies were successfully prepared. Using mouse anti-PNA polyclonal antibody as the capture antibody and rabbit anti-PNA polyclonal antibody as the detection antibody, a novel sandwich enzyme-linked immunosorbent assay (ELISA) was developed to quantify PNA. The detection limit of the ELISA method was low (0.49 ng/mL), and the linear dynamic range was between 0.78 and 100 ng/mL. The recovery ranged from 93.86% to 139.3%, whereas the intra- and inter-assay coefficients of variation were less than 10.25% and 12.06%, respectively. Sample analysis verified this method as a reliable tool for the detection of PNA in processed foods.
研究领域:
免疫学
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食品科学
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DOI
高影响因子
Analytical Methods
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2015
·Vol. 7
·期 15
·6190-6197
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IF 1.821
Xiaoling Wu,Dezhao Kong,Shanshan Song,Liqiang Liu,Hua Kuang,Chuanlai Xu
A pair of sensitive monoclonal antibodies against both Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola were generated by immunizing mice and cell fusion techniques. The sandwich enzyme-linked immunosorbent assay method and immunochromatographic test strip were developed based on newly generated monoclonal antibodies for detection in rice samples. The lower limit of detection of the sandwich ELISA method for X. oryzae pv. oryzae was 1.0 × 104 cfu mL−1. The cut-off values for the immunochromatographic test strip for X. oryzae pv. oryzae and X. oryzae pv. oryzicola in rice samples were 3.3 × 105 cfu mL−1 and 1 × 106 cfu mL−1, respectively. The quantitative sandwich ELISA method and rapid immunochromatographic test strip assay could be useful for food safety inspections.
研究领域:
植物病理学
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微生物学
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食品安全
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DOI
高影响因子
ANALYTICAL CHEMISTRY
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2021
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IF 6.986
Hailing Zheng,Hailiang Yang,Yang Zhou,Tianxiao Li,Qinglin Ma,Bing Wang,Qin Fang,Haixiang Chen
The extraction and identification of silk residues in tombs is of great significance for studying the distribution and spread of early silk. However, the complex organic matter in the tomb hinders the accurate identification of silk. In this study, a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) based on immunomagnetic beads (IMBs) was developed for the rapid enrichment and detection of silk residues. The double-antibody sandwich ELISA method established by pairing the IMBs prepared by the silk fibroin monoclonal antibody SF-3 and the silk fibroin monoclonal-labeled antibody bio-SF-1 had the highest detection sensitivity, with a linear detection range of 10 to 104 ng mL–1 and a detection limit of 5.12 ng mL–1. This method was excellent in the extraction and analysis of silk residues from archaeological imprints and soil samples and successfully identified silk residues in samples at the final stage of silk degradation (physical invisible silk). The proteomics analysis results demonstrated the feasibility and practicability of this method.
研究领域:
考古学
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生物化学
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材料科学
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DOI
PubMed
高影响因子
RSC Advances
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2023
·Vol. 13
·期 16
·10503-10507
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IF 3.9
Chenlong Jiang,Xiangde Lai,Feng Han,Zhijun Gao,Haixia Yang,Xuan Zhao,Huajie Pang,Bin Qiao,Hua Pei,Qiang Wu
Heart failure (HF) is a major public health problem triggered by heart circulation disorders. Early detection and diagnosis are conducive to the prevention and treatment of HF. Hence, it is necessary to establish a simple and sensitive method to monitor the diagnostic biomarkers of HF. The N-terminal B-type natriuretic peptide precursor (NT-proBNP) is acknowledged as a sensitive biomarker. In this study, a visual detection method for NT-proBNP was developed based on the oxidized 3,3′,5,5′-tetramethylbenzidine (TMB2+)-mediated etching of gold nanorods (AuNRs) and double-antibody-sandwich ELISA. The etching color for different amounts of NT-proBNP was obvious and significant differences could be ascertained based on the blue-shift of the longitudinal localized surface plasmon resonance (LLSPR) of the AuNRs. The results could be observed by the naked eye. The constructed system showed a concentration range from 6 to 100 ng mL−1 and a low detection limit of 6 ng mL−1. This method exhibited negligible cross-reactivity toward other proteins, and the recoveries of the samples ranged from 79.99% to 88.99%. These results demonstrated that the established method is suitable for the simple and convenient detection of NT-proBNP.
研究领域:
生物标志物检测
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纳米技术
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF DAIRY SCIENCE
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2016
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IF 2.408
Shan Shan,Daofeng Liu,Qi Guo,Songsong Wu,Rui Chen,Kai Luo,Liming Hu,Yonghua Xiong,Weihua Lai
In this study, cascade signal amplification in ELISA involving double-antibody sandwich ELISA and indirectly competitive ELISA was established to sensitively detect Escherichia coli O157:H7. In the double-antibody sandwich ELISA, a complex was formed comprising anti- E. coli O157:H7 polyclonal antibody, E. coli O157:H7, biotinylated anti- E. coli O157:H7 monoclonal antibody, streptavidin, and biotinylated β-lactamase. Penicillin solution was then added into the ELISA well and hydrolyzed by β-lactamase. Afterward, the penicillin solution was transferred to indirectly competitive ELISA. The concentration of penicillin can be sensitively detected in indirectly competitive ELISA. In the cascade signal amplification system, increasing the amount of added E. coli O157:H7 resulted in more β-lactamase and less penicillin. The detection sensitivity of E. coli O157:H7, which was 20 cfu/mL with the cascade signal amplification in ELISA, was 1,000-fold higher than that of traditional ELISA. Furthermore, the novel method can be used to detect E. coli O157:H7 in milk (2 cfu/g). Therefore, this new signaling strategy will facilitate analyses of highly sensitive foodborne pathogens.
研究领域:
微生物学
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食品科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2022
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IF 6.911
Yarong Ren,Juan Wei,Ye Wang,Peng Wang,Yanwei Ji,Baoyuan Liu,Jianlong Wang,Gualberto González-Sapienza,Yanru Wang
Salmonella are major pathogens that cause foodborne diseases. In this work, a broad-spectrum Salmonella nanobody-01 (Nb-01) was isolated and applied in the development of a streptavidin-bridged sandwich ELISA (SAB-ELISA) for simultaneously identifying five Salmonella serovars , including Salmonella Enteritidis ( S. Enteritidis), Salmonella Typhimurium ( S. Typhimurium), Salmonella London (S. London), Salmonella Paratyphi B (S. Paratyphi B) and Salmonella Hadar (S. Hadar). In this work, streptavidin (SA) was utilized as a scaffold to directionally immobilize biotinylated nanobody (BiNb) and Salmonella was detected by phage-displayed nanobodies. The SAB-ELISA can be accomplished within 180 min with a limit of detection (LOD) of 6.31 × 10 3 colony forming units (CFU) mL −1 , 9.15 × 10 3 CFU mL −1 , 4.23 × 10 3 CFU mL −1 , 7.31 × 10 3 CFU mL −1 and 7.25 × 10 3 CFU mL −1 towards S. Typhimurium, S. Enteritidis, S. London, S. Paratyphi B and S. Hadar, respectively. In comparison of sandwich ELISA by passive immobilization of Nb-01 on polystyrene plate, the sensitivity was increased by around 6-fold, which confirmed the enhanced immobilization efficacy of SAB-ELISA. Furthermore, the feasibility of the assay for S. Typhimurium determination in actual samples was evaluated, showing excellent recovery, inter-day and intra-day precision.
研究领域:
微生物学
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免疫学
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食品科学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
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