Hapten synthesis, monoclonal antibody production and immunoassay development for direct detection of 4-hydroxybenzehydrazide in chicken, the metabolite of nifuroxazide

Mari, Ghulam Mujtaba; Li, Hongfang; Dong, Baolei; Yang, Huijuan; Talpur, Aisha; Mi, Jiafei; Guo, Liuchuan; Yu, Xuezhi; Ke, Yuebin; Han, Diangang; Wang, Zhanhui

doi:10.1016/j.foodchem.2021.129598

Cited by 22 publications

(8 citation statements)

References 27 publications

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“…To enhance the assay sensitivity and stability, three parameters were optimized. The A max /IC 50 ratio was used as the index for evaluating the influence of each condition; a higher ratio demonstrates a higher level of sensitivity in the assay [ 44 , 45 , 46 , 47 ]. The optimized results are shown in Figure 3 .…”

Section: Resultsmentioning

confidence: 99%

A Sensitive Monoclonal-Antibody-Based ELISA for Forchlorfenuron Residue Analysis in Food Samples

et al. 2022

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In this study, forchlorfenuron (CPPU) was coupled with succinic anhydride to yield a CPPU hapten (CPPU-COOH), and a high-affinity monoclonal antibody (mAb) that can specifically recognize CPPU was produced. Using this mAb as a recognition reagent, a sensitive indirect competitive enzyme-linked immunosorbent assay (icELISA) for CPPU was optimized, which exhibits an IC50 of 1.04 ng/mL, a limit of detection of 0.16 ng/mL, and a linear range of 0.31–3.43 ng/mL for CPPU. Cross-reactivity percentages with six analogues were all below 6%. The average recovery rates for cucumber and orange samples were from 85.23% to 119.14%. The analysis results of this icELISA showed good consistency with those from liquid chromatography mass spectrometry. These results suggest that the proposed icELISA provides a sensitive, specific, and reliable strategy for CPPU detection in food samples.

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Section: Resultsmentioning

confidence: 99%

A Sensitive Monoclonal-Antibody-Based ELISA for Forchlorfenuron Residue Analysis in Food Samples

et al. 2022

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“…Hapten design is key to generating excellent antibody performances against small molecules [25]. Generally, haptens should mimic the target molecule in terms of size, shape, electronic properties, and insert functional groups such as, carboxyl, amino, and sulfhydryl groups for carrier protein coupling [16,17]. In principle, there are two ways to synthesize PbTx-2 haptens.…”

Section: Hapten Design Synthesis and Conjugate Preparationmentioning

confidence: 99%

“…It is accepted that the coupling ratio is an important factor affecting generated antibodies; higher coupling ratios could induce higher antibody titers [17]. In the literature, appropriate coupling ratios ranged from 3 to 15 [16,17,21]. The coupling ratios of PbTx-2-CMO-KLH/BSA and PbTx-2-HS-KLH/BSA were relatively low due to the lower reaction molar ratio (hapten to carrier protein).…”

Section: Hapten Design Synthesis and Conjugate Preparationmentioning

confidence: 99%

“…In principle, aldehyde groups from PbTx-2 may be conjugated with a spacer arm, such as carboxymethoxylamine hemihydrochloride (CMO) and/or aminobenzoic acid. In addition, molecular modeling has become a powerful tool in guiding and improving hapten design strategies [15][16][17][18][19]. Therefore, we explored and developed novel haptens using molecular modeling to prepare mAbs against brevetoxins.…”

Section: Introductionmentioning

confidence: 99%

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Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay

Zhang

Ding

Zhang

et al. 2021

Foods

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The consumption of shellfish contaminated with brevetoxins, a family of ladder-frame polyether toxins formed during blooms of the marine dinoflagellate Karenia brevis, can cause neurotoxic poisoning, leading to gastroenteritis and neurotoxic effects. To rapidly monitor brevetoxin levels in oysters, we generated a broad-spectrum antibody against brevetoxin 2 (PbTx-2), 1 (PbTx-1), and 3 (PbTx-3) and developed a rapid indirect competitive enzyme-linked immunosorbent assay (icELISA). PbTx-2 was reacted with carboxymethoxylamine hemihydrochloride (CMO) to generate a PbTx-2-CMO hapten and reacted with succinic anhydride (HS) to generate the PbTx-2-HS hapten. These haptens were conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen reagents, respectively, using the active ester method. After immunization and cell fusion, a broad-spectrum monoclonal antibody (mAb) termed mAb 1D3 was prepared. The 50% inhibitory concentration (IC50) values of the icELISA for PbTx-2, PbTx-1, and PbTx-3 were 60.71, 52.61, and 51.83 μg/kg, respectively. Based on the broad-spectrum mAb 1D3, an icELISA was developed to determine brevetoxin levels. Using this approach, the limit of detection (LOD) for brevetoxin was 124.22 μg/kg and recoveries ranged between 89.08% and 115.00%, with a coefficient of variation below 4.25% in oyster samples. These results suggest that our icELISA is a useful tool for the rapid monitoring of brevetoxins in oyster samples.

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“…To realize coupling to macromolecular substances, the aldicarb hapten molecule must have an active group (such as -NH 2 , -COOH, -OH, and -SH) that can covalently bind to the carrier, and to enhance the performance of the resulting antibody, the spacer arms of the hapten should be of a certain length (20). Current studies have found that the length and composition of the spacer arm affect the ability of the antibody to recognize the analyte, that is, the affinity of the antibody for the analyte, because the spacer arm affects the properties and structures of small pesticide molecules in artificial antigen molecules (21)(22)(23).…”

Section: Introductionmentioning

confidence: 99%

Hapten designs based on aldicarb for the development of a colloidal gold immunochromatographic quantitative test strip

Shen

Yue

et al. 2022

Front. Nutr.

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The common carbamate insecticide aldicarb is considered one of the most acutely toxic pesticides. Herein, rational design was used to synthesize two haptens with spacers of different carbon chain lengths. The haptens were then used to immunize mice. The antibodies obtained were evaluated systematically, and a colloidal gold immunochromatographic strip was developed based on an anti-aldicarb monoclonal antibody. The 50% inhibition concentration and linear range of anti-aldicarb monoclonal antibody immunized with Hapten 1 were 0.432 ng/mL and 0.106–1.757 ng/mL, respectively. The cross-reactivities for analogs of aldicarb were all <1%. The limit of detection of the colloidal gold immunochromatographic strip was 30 μg/kg, and the average recoveries of aldicarb ranged from 80.4 to 110.5% in spiked samples. In the analysis of spiked samples, the test strip could accurately identify positive samples detected by the instrumental method in the GB 23200.112-2018 standard but produced some false positives for negative samples. This assay provides a rapid and accurate preliminary screening method for the determination of aldicarb in agricultural products and environments.

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Hapten synthesis, monoclonal antibody production and immunoassay development for direct detection of 4-hydroxybenzehydrazide in chicken, the metabolite of nifuroxazide

Cited by 22 publications

References 27 publications

A Sensitive Monoclonal-Antibody-Based ELISA for Forchlorfenuron Residue Analysis in Food Samples

A Sensitive Monoclonal-Antibody-Based ELISA for Forchlorfenuron Residue Analysis in Food Samples

Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay

Hapten designs based on aldicarb for the development of a colloidal gold immunochromatographic quantitative test strip

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