A sandwich enzyme immunoassay for wheat gliadin

Troncone, Riccardo; Vitale, Mario; A, Donatiello; Farris, Evelina; Rossi, Guido; Auricchio, Salvatore

doi:10.1016/0022-1759(86)90498-9

Cited by 41 publications

(12 citation statements)

References 8 publications

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“…All three assays are less sensitive than the two-site assays of Windemann et al (1982), Troncone et al (1986) and Ayob et al (1988). The reason for this may be that these assays employed polyclonal antisera throughout and were probably of broader specificity in terms of the gliadin sub-fractions recognized.…”

Section: Discussionmentioning

confidence: 88%

A two‐site enzyme‐linked immunosorbent assay for wheat gliadins

Mills¹,

Spinks²,

Morgan³

1989

Food and Agricultural Immunology

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A two site enzyme-linked immunosorbent assay for wheat gliadins has been developed. Polyclonal antibodies from chicken eggs were used as the capture agent adsorbed to the surface of the microtitration plate. Murine monoclonal antibodies bound to captured gliadin were detected by addition of enzyme-labelled, species-specific antibodies. The assay was developed for the purposes of screening monoclonal antibodies specific for conformational epitopes on gliadins. The properties of a panel of 6 anti-gliadin antibodies in the assay were assessed and compared to properties exhibited under conditions where conformational epitopes were less likely to be present. Three of the monoclonal antibodies appeared to be specific for conformational epitopes.

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Section: Discussionmentioning

confidence: 88%

A two‐site enzyme‐linked immunosorbent assay for wheat gliadins

Mills¹,

Spinks²,

Morgan³

1989

Food and Agricultural Immunology

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“…Among the different methods published, Skerritt and Smith (1985) developed a method based on MAbs that allows the evaluation of thermally treated samples, but has only limited sensitivity. Troncone et al (1986) developed a capture ELISA with PAbs that exhibits a high detectability, but it showed cross-reactivity with non-toxic prolamins from rice and maize. Friis (1988) using PAbs developed a competitive ELISA which in spite of being able to detect up to 10 ng gliadin/ml cannot be used on thermally processed food.…”

Section: Discussionmentioning

confidence: 99%

“…Using some antibodies, cross-reactivity with proteins non-toxic for coeliac patients-leading to false-positive results-have also been documented in quantitative methods (e.g. Troncone et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system

Chirdo¹,

Añón

Fossati

1998

Food and Agricultural Immunology

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Optimization of three enzyme immunoassays of very high sensitivity using three antiprolamin monoclonal antibodies (MAbs) (13B4, 11C4 and 12A1) is presented here. These MAbs are specific for those prolamins toxic for coeliac patients, as determined by immunoblotting analysis. Biotinylated MAbs were used in two of the assays. In a competitive ELISA, the binding of each biotinylated MAb to a gliadin-coated solid phase was inhibited by gliadin in the fluid phase. The best result was obtained using the biotinylated MAbl3B4 (detection limit: 20 ng ml-1). With regard to capture ELISA, we tested the performance of the three MAbs. In this sandwich ELISA, the MAb used for antigenic capture was the same as that used as secondary biotinylated antibody. The MAbl2Al had the best performance (detection limit: 1 ng ml-1). The use of biotin-labelled gliadin in a quantitative immunoassay with a detection limit of 5 ng ml-1 is also reported. This assay involves an antigenic capture using the MAbl2Al followed by a competition between biotinylated and non-biotinylated gliadin. We have found the use of the streptavidin-biotin interaction as signal amplification system to be very useful. This technique, as far as we know, has not been previously reported for gliadin quantification.

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“…Numerous commercial ELISA kits are available on the market from several suppliers (Jayasena et al., 2015; Monaci & Visconti, 2010; Panda et al., 2010). Available commercial wheat ELISA kits may differ in format, specificity, and sensitivity depending on the manufacturer, while these ELISA methods are only specific for a single allergen, such as α‐amylase inhibitor, gluten, gliadin (Scherf, 2017; Scherf & Poms, 2016; Troncone et al., 1986; Verity et al., 1999). Nevertheless, wheat and its derivatives are usually consumed after being exposed to certain degrees of processing treatments, which would cause the denaturation and the aggregation of some wheat proteins due to intra‐ and inter‐molecular interactions of the proteins in the complex food matrix (Nowak‐Wegrzyn & Fiocchi, 2009).…”

Section: Introductionmentioning

confidence: 99%

A sensitive sandwich enzyme‐linked immunosorbent assay (sELISA) targeted multiple wheat protein fractions for the detection of several cereal grains in processed foods

Zhao

Khan

Chen

et al. 2022

Journal of Food Science

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Wheat allergy has become a global public health and food safety concern; however, there is no accessible cure for wheat allergy. The complete exclusion of wheat‐containing foods and environmental exposure is the most efficient allergy management to avoid the adverse reactions, which can be severe and occasionally life threatening. Therefore, the assay for accurate detection of wheat residues is demanded urgently for appropriate labeling guidelines and consumer safety. Thus, a sandwich enzyme‐linked immunosorbent assay (sELISA) targeting multiple wheat protein fractions was fabricated in the present study. The results showed that the limit of detection (LOD) and limit of quantitation (LOQ) of the constructed sELISA were 0.25 and 0.5 µg/g with high specificity for wheat. No cross‐reactivity was observed in 32 foods or food ingredients tested, except barley and rye. The developed sELISA can also discriminate against many commercial foods containing declared or undeclared wheat residues except for Chinese yellow wine. Furthermore, high heat also can obtain a higher level of proteins extracted with corresponding enhanced detectability up to 100°C from heated samples and 160 °C in baked samples. Practical Application Wheat is the most common food ingredient and wildly applied in various processed foods. However, wheat can cause severe and life‐threatening symptoms in some allergic patients and must be labeled and tested accurately to protect those with a wheat allergy. Developing a new test assay can serve as a powerful tool for food manufacturers and regulatory agencies to accurately quantify wheat residues in processed foods and ensure their absence due to unintended contamination.

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A sandwich enzyme immunoassay for wheat gliadin

Cited by 41 publications

References 8 publications

A two‐site enzyme‐linked immunosorbent assay for wheat gliadins

A two‐site enzyme‐linked immunosorbent assay for wheat gliadins

Development of high‐sensitive enzyme immunoassays for gliadin quantification using the streptavidin‐biotin amplification system

A sensitive sandwich enzyme‐linked immunosorbent assay (sELISA) targeted multiple wheat protein fractions for the detection of several cereal grains in processed foods

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