Sensitive Monoclonal Antibody-Based Immunoassays for Kresoxim-methyl Analysis in QuEChERS-Based Food Extracts

A monoclonal antibody against carazolol (CRZ), 2F5, was generated for the construction of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and an immunochromatographic strip for detection of CRZ in milk and pig kidney samples. The antibody had a half-maximal inhibitory concentration of 0.29 ng/ mL; its limit of detection was 0.068 ng/mL, and its linear range of detection was 0.137-0.638 ng/mL. For CRZ in milk samples and porcine kidney samples, the detection limits for this strip were 0.25 and 2.5 ng/mL, respectively. Analysis of CRZ in milk samples showed that the results obtained by immunochromatographic strip were consistent with those obtained by ic-ELISA. Therefore, the CG immunoassay is a sensitive screening method for semiquantitative and qualitative detection of residual CRZ in milk and porcine kidney samples.

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“…The CR values were calculated as follows: CR (%) = (IC 50 of CRZ/IC 50 of competitor) × 100% (Mercader et al, 2014).…”

Section: Production Of Monoclonal Antibodiesmentioning

confidence: 99%

Development of an ic-ELISA and colloidal gold strip for the detection of the beta-blocker carazolol

Zhou

Kuang

Liu

2020

Food and Agricultural Immunology

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“…In the references, the minimum detectable mass fraction and recovery rate were 0.02 mg/kg and 81.8-99.15% by HPLC, respectively (Chen et al 2011;Wan et al 2011), 0.005 mg/kg and 80.2-103% by GC (Zhang et al 2008), and 0.001-0.02 mg/kg and 76-106% by LC-ESI-MS-MS (Anna et al 2004). In our experiment, IC 50 values were 14.7 and 22.9 µg/ml, and the limits of detection were 0.0044 and 0.014 g/ml, respectively, for kresoxim-methyl and trifloxystrobin, which were below the common maximum residue limit, 0.05 g/ml (Mercader et al 2014a). According to the determined results for spiked orange samples, the recovery rates of the new ELISA assay were 82.73-104.35% for kresoxim-methyl and 91.1-92.43% for trifloxystrobin, which were the values consistent with those of the above spectrometry examination.…”

Section: Discussionmentioning

confidence: 48%

“…In Europe and in the USA, MRLs for kresoxim-methyl range between 0.05 and 1.5 mg/kg Czech J. Food Sci., 34, 2016(5): 429-438 doi: 10.17221/275/2015 in most fruits and vegetables (Mercader et al 2014a), and for trifloxystrobin they range from 0.05 mg/kg to 30 mg/kg in authorised crops (Mercader et al 2014b). The most stringent limits of trifloxystrobin and kresoxim-methyl concentrations in oranges are 0.02-2.0 and 0.05 mg/kg, respectively, in the European Union, and 0.05-10.0 and 0.03 mg/kg in Japan (Campillo et al 2010).…”

Section: Kresoxim-methyl Methyl(e)-methoxyimino[α-(o-tolyloxy)-o-tolmentioning

confidence: 99%

Enzyme-linked immunosorbent assay for simultaneous detection of two fungicides kresoxim-methyl and trifloxystrobin in oranges

Yang¹,

Zhu²,

Liu³

et al. 2016

Czech J. Food Sci.

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Yang W., Zhu J., Liu M.Q., Dai X.J. (2016): Enzyme-linked immunosorbent assay for simultaneous detection of two fungicides kresoxim-methyl and trifloxystrobin in oranges. Czech J. Food Sci., 34: 429-438.To assemble an indirect competitive enzyme-linked immunosorbent assay (ELISA) for estimating two strobilurin fungicides at the same time, the hapten was synthesised which contained the common active group, (E)-2-(2-bromophenyl)-2-(methoxyimino) acetic acid methyl ester (OEBr) in kresoxim-methyl and trifloxystrobin. The immunogen and coating antigen were respectively prepared through conjugating the above-mentioned hapten with BSA and OVA by the mixed anhydride and activated ester methods, and polyclonal antibodies were produced by immunised rabbits. An enzyme-linked immunosorbent assay was developed for simultaneous detection of kresoxim-methyl and trifloxystrobin. In ELISA, the antiserum showed high affinity and sensitivity to kresoxim-methyl and trifloxystrobin, and their IC 50 value and detection limit (expressed as IC 10 ) were 14.7 and 0.0044 µg/ml, respectively, for kresoxim-methyl, and 22.9 and 0.014 µg/ml, respectively, for trifloxystrobin. The cross-reaction rate was below 0.1% for other strobilurin fungicides. Recovery study of ELISA from spiked samples of homogenised peeled oranges (final concentrations of 100, 10, and 1 µg/ml) resulted in recovery levels in the range of 82-104%.

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“…This pretreatment method involves only three steps: extraction of sample with organic solvent, transfer of the extract to a d-SPE tube and cleanup by d-SPE. The d-SPE uses less sorbent and equipment while also saving time, effort and money, thus making it more applicable in the sample preparation procedure before analysis by immunoassay323334.…”

Section: Resultsmentioning

confidence: 99%

Development and comparative study of chemosynthesized antigen and mimotope-based immunoassays for class-specific analysis of O,O-dimethyl organophosphorus pesticides

Zhao

Wang

Han

et al. 2016

Sci Rep

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The multi-residue determination of organophosphorus pesticides (OPs) is an important task due to the wide application and high toxicity of OPs. However, there is no promising immunoassay to monitor the multi-residue of O,O-dimethyl OPs. In this study, a monoclonal antibody (mAb) against a generic hapten of O,O-dimethyl OPs (O,O-dimethyl O-(3-carboxyphenyl)phosphorothioate) was prepared. To develop an effective class-specific immunoassay, two strategies were performed to select the appropriate coating antigen or competing antigen. On the one hand, a total of 20 haptens were chemosynthesized, attached to ovalbumin for use as coating antigen candidates, and selected by direct competitive ELISA (dcELISA). As a second strategy, mimotopes of the mAb were selected from a random phage-display peptide library by panning, and the optimum mimotope was expressed as a fusion protein and biotinylated in vitro. Based on the selected chemosynthesized coating antigen and the biotinylated mimotope fusion protein, two sensitive broad-specificity dcELISAs were developed. The sensitivity, selectivity and practicability of the two immunoassays were compared. The results demonstrated that both methods showed similar selectivity and sensitivity and were reliable for O,O-dimethyl OP residues screening. However, the screening operation of mimotopes was much simpler and safer compared to the preparation of chemosynthesized coating antigens.

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Sensitive Monoclonal Antibody-Based Immunoassays for Kresoxim-methyl Analysis in QuEChERS-Based Food Extracts

Cited by 7 publications

References 14 publications

Development of an ic-ELISA and colloidal gold strip for the detection of the beta-blocker carazolol

Development of an ic-ELISA and colloidal gold strip for the detection of the beta-blocker carazolol

Enzyme-linked immunosorbent assay for simultaneous detection of two fungicides kresoxim-methyl and trifloxystrobin in oranges

Development and comparative study of chemosynthesized antigen and mimotope-based immunoassays for class-specific analysis of O,O-dimethyl organophosphorus pesticides

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