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A paper-based competitive lateral flow immunoassay for multi ?-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles.

ABSTRACT: An ultrasensitive paper based lateral flow assay is described for rapid and simultaneous fluorometric detection of several ?-agonists including clenbuterol and its chemical analogues (mabuterol, brombuterol, cimaterol, cimbuterol, bromchlorbuterol and banbuterol). A nonspecific monoclonal antibody (mAb) against clenbuterol and its analogues was prepared and employed in a competitive immunoassay where mAb conjugated to fluorescent nanoparticles and free ?-agonists compete for the binding sites. This enables rapid screening for the 7 ?-agonists in a single run that takes about 8 min. Detection limits for the seven ?-agonists are <50 pg g-1 of pork. Recoveries ranged from 69.5% to 102.4%, and relative standard deviations were ±15%. The assay was applied to the analysis of both using spiked and unspiked pork for ?-agonists, and the results compare well to those obtained by HPLC-MS. Graphical abstractSchematic presentation of an ultra sensitive fluorescent nanoparticle based paper based assay for rapid detection of multi ?-agonists in pork tissue.

SUBMITTER: Wang R

PROVIDER: S-EPMC5823949 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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A paper-based competitive lateral flow immunoassay for multi β-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles.

Wang Ruiguo R Zhang Wei W Wang Peilong P Su Xiaoou X

Mikrochimica acta 20180222 3

An ultrasensitive paper based lateral flow assay is described for rapid and simultaneous fluorometric detection of several β-agonists including clenbuterol and its chemical analogues (mabuterol, brombuterol, cimaterol, cimbuterol, bromchlorbuterol and banbuterol). A nonspecific monoclonal antibody (mAb) against clenbuterol and its analogues was prepared and employed in a competitive immunoassay where mAb conjugated to fluorescent nanoparticles and free β-agonists compete for the binding sites. T ...[more]

PMID: 29503465

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Project description:A fluorescent immunoassay based on europium nanoparticles (EuNPs-FIA) was developed for the simultaneous detection of antibiotic residues, solving the problems of single target detection and low sensitivity of traditional immunoassay methods. In the EuNPs-FIA, EuNPs were used as indictive probes by binding to anti-tetracyclines monoclonal antibodies (anti-TCs mAb), anti-sulphonamides monoclonal antibodies (anti-SAs mAb) and anti-fluoroquinolones monoclonal antibodies (anti-FQs mAb), respectively. Different artificial antigens were assigned to different regions of the nitrocellulose membrane as capture reagents. The EuNPs-FIA allowed for the simultaneous detection of three classes of antibiotics (tetracyclines, fluoroquinolones and sulphonamides) within 15 min. It enabled both the qualitative determination with the naked eye under UV light and the quantitative detection of target antibiotics by scanning the fluorescence intensity of the detection probes on the corresponding detection lines. For qualitative analysis, the cut-off values for tetracyclines (TCs), fluoroquinolones (FQs) and sulphonamides (SAs) were 3.2 ng/ml, 2.4 ng/ml and 4.0 ng/ml, respectively, which were much lower than the maximum residue limit in food. For quantitative analysis, these ranged from 0.06 to 6.85 ng/ml for TCs, 0.03-5.14 ng/ml for FQs, and 0.04-4.40 ng/ml for SAs. The linear correlation coefficients were higher than 0.97. The mean spiked recoveries ranged from 92.1 to 106.2% with relative standard deviations less than 8.75%. Among them, the three monoclonal antibodies could recognize four types of TCs, seven types of FQs and 13 types of SAs, respectively, and the detection range could cover 24 antibiotic residues with different structural formulations. The results of the detection of antibiotic residues in real samples using this method were highly correlated with those of high performance liquid chromatography (R 2 > 0.98). The accuracy and precision of the EuNPs-FIA also met the requirements for quantitative analysis. These results suggested that this multiplex immunoassay method was a promising method for rapid screening of three families of antibiotic residues.

Dataset Information

A paper-based competitive lateral flow immunoassay for multi ?-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles.

Publications

A paper-based competitive lateral flow immunoassay for multi β-agonist residues by using a single monoclonal antibody labelled with red fluorescent nanoparticles.

Similar Datasets

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