Sensitivity and specificity enhanced enzyme-linked immunosorbent assay by rational hapten modification and heterogeneous antibody/coating antigen combinations for the detection of melamine in milk, milk powder and feed samples

Cao, Biyun; Yang, Hong; Song, Juan; Chang, Huafang; Li, Shuqun; Deng, Anping

doi:10.1016/j.talanta.2013.05.009

Cited by 50 publications

(28 citation statements)

References 30 publications

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“…40,42–44 Therefore, additional haptens were used in the development of heterologous immunoassays (Haptens 5and 6). Fipronil analogue, compound 2 (Scheme 1), and 2,6-dichloro-4-(trifluoromethyl)aniline were attached to protein without additional modification through the amine group.…”

Section: Resultsmentioning

confidence: 99%

Development of an Immunoassay for the Detection of the Phenylpyrazole Insecticide Fipronil

Vasylieva

Ahn

Barnych

et al. 2015

Environ. Sci. Technol.

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Phenylpyrazole insecticides such as fipronil have been used as replacements for organophosphates. The wide application of fipronil raises concern about environmental contamination and risk for fish, birds, other non-targeted beings and human health. A sensitive, competitive indirect heterologous enzyme-linked immunosorbent assay (ELISA) was developed. Antibodies with different specificities to fipronil and its metabolites were produced. Two ELISAs having IC50 values of 0.58 ± 0.06 and 2.6 ± 0.4 ng/mL were developed. Design of different haptens and coating antigens resulted in two assays with distinct cross-reactivity patterns for structurally related compounds: 96%, 38% and 101% vs 39%, 1.4% and 25% for fipronil-sulfide, fipronil-detrifluoromethylsulfonyl and fipronil-desulfinyl, respectively. Performance of the immunoassays was demonstrated by a recovery study from spiked water, human serum and urine matrices, giving recovery values in the range of 85–111% for different concentrations. The assays demonstrated good correlation in fipronil recovery with conventional LC-MS/MS analysis. The generic assay 2265 has the sensitivity to measure fipronil and its analogs in serum at levels relevant for exposure monitoring. The assays were used to analyze human urine samples obtained from exposure studies and serum samples from rats treated with fipronil-containing diet.

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Section: Resultsmentioning

confidence: 99%

Development of an Immunoassay for the Detection of the Phenylpyrazole Insecticide Fipronil

Vasylieva

Ahn

Barnych

et al. 2015

Environ. Sci. Technol.

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“…Cao reported previously a method for the production of a monoclonal antibody which demonstrated a higher specicity towards melamine. 1 However, the ability to screen for cyromazine should not be viewed as a drawback. The immunoassay developed in this report is designed as a screening tool for food safety purposes.…”

Section: Discussionmentioning

confidence: 99%

“…The white solid was then dried by vacuum (13.2 g, 93.1%). 1 Aer obtaining CAAT, AETT was synthesized from it by Zhou's method. 17 Briey, ethylenediamine (3.1 g) was dissolved in water, followed by the addition of CAAT (1.45 g).…”

Section: Synthesis Of the Coating Antigenmentioning

confidence: 99%

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A simple, highly sensitive, high throughput and organic solvent-free screening method for melamine by microsphere-based flow cytometry immunoassay

Tsoi

Wong

2015

Anal. Methods

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In recent years, the safety problem of infant formula powder has provoked panic among Chinese citizens.One of the most notorious incidents was the 2008 Chinese milk scandal which harmed thousands of infants. Therefore the development of a fast, high throughput screening method towards melamine and other harmful chemicals is of paramount importance. Microsphere-based flow cytometry is a new multiplexing immunochemical method for food-testing. Through competition between the free analyte in the sample and analyte-coupled microspheres, the content of the analyte can be indirectly quantified by measuring the fluorescence emitted from labelled antibodies. This developed method of high sensitivity (limit of detection ¼ 0.70 ng ml À1 ) has already been validated and successfully applied to screen for melamine in various brands of infant formula powder analysis fulfilling the legislated tolerance level in a simple, fast, high throughput and organic solvent-free manner.

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“…Crosslinking reagents have been used to polymerize those functional groups into macromolecules to obtain haptenic polymers which will be used to immunize animal for antibodies production. The non-immunogenic MA has been polymerized using glutaraldehyde to reach a preferred complete antigen size at least of 4 kDa [78,79]. However, in vivo produced antibodies still have experimental limitations in their applications, mainly non-specific binding that can lead to cross-reactivity and high background.…”

Section: Merits Of Aptamer-based Techniquesmentioning

confidence: 99%

Current Advances and Prospects on Implementation of Highly Sensitive Aptamer-based Dual System for Melamine Detection: New Promising Tool of Great Affinity

Ndikubwimana¹

2015

J Bioprocess Biotech

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Sensitivity and specificity enhanced enzyme-linked immunosorbent assay by rational hapten modification and heterogeneous antibody/coating antigen combinations for the detection of melamine in milk, milk powder and feed samples

Cited by 50 publications

References 30 publications

Development of an Immunoassay for the Detection of the Phenylpyrazole Insecticide Fipronil

Development of an Immunoassay for the Detection of the Phenylpyrazole Insecticide Fipronil

A simple, highly sensitive, high throughput and organic solvent-free screening method for melamine by microsphere-based flow cytometry immunoassay

Current Advances and Prospects on Implementation of Highly Sensitive Aptamer-based Dual System for Melamine Detection: New Promising Tool of Great Affinity

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