Quantification of profilins by a monoclonal antibody-based sandwich ELISA

Asturias, Juan A.; Arilla, Maite; Aguirre, Manuel A.; Gómez-Bayón, N; Martı́nez, Alberto; Palácios, Ricardo; Sánchez-Gascón, F; Martı́nez, Jorge

doi:10.1016/s0022-1759(99)00115-5

Cited by 26 publications

(23 citation statements)

References 31 publications

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“…It was very surprising to see that although the mice were boosted with rPru p 3, the reactions to the antibodies were greater with nPru p3. A very similar phenomenon also has been reported when quantifying peach profilin, demonstrating that the use of a recombinant protein isoform as standard might overestimate the profilin content of samples (Asturias et al 1999). One explanation may be that recombinant proteins are not correctly folded as natural Pru p 3, and also lack of natural ligands, such as lipids which may affect affinity (Bublin et al 2014).…”

Section: Discussionsupporting

confidence: 60%

“…The latter is the most common type of immunoassay for detecting potential food allergens and has been developed for several of them, with numerous test kits made commercially available in this format during the last decade. Sandwich ELISA has been used to quantify the peach allergen (Asturias et al 1999;Ahrazem et al 2007). In most sandwich ELISAs, a monoclonal antibody is usually used as the first antibody and a polyclonal as the detection antibody (Carnes et al 2002).…”

Section: Introductionmentioning

confidence: 99%

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Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

Zhou

Yang

et al. 2015

Food Anal. Methods

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Fruit is one of the most commonly reported food allergy sources in China, and peach lipid transfer protein (Pru p 3) has been identified as the major allergen inducing systematic symptoms. Crude allergen extracts and single component allergens have been used in food allergy diagnosis and immunotherapy. Reliable and sensitive analytical methods to quantify Pru p 3 content in fruit will help to identify lowallergenic cultivars among the abundant peach genetic resources in China. In this study, we developed a sensitive sandwich ELISA method to measure Pru p 3 in peach fruit based on two monoclonal antibodies with high sensitivity and linearity. Significant variability was observed in peach cultivars, with a much higher Pru p 3 level in peach peel than in pulp. This method will be very useful to select peach varieties with low Pru p 3 content and as a diagnostic product to detect hidden allergens in processed food.

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Section: Discussionsupporting

confidence: 60%

Section: Introductionmentioning

confidence: 99%

Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

Zhou

Yang

et al. 2015

Food Anal. Methods

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“…In our study, histamine release curves with purified profilins and orange extract had almost identical shapes, indicating that the whole allergenic potency of the extract was represented by profilin. Concentration differences between extract and purified protein are due to the fact that profilin represents a minor protein constituent in plants, for example, 0.015 to 0.02% of total protein in various pollens [34]. Therefore, we conclude that it is very likely that Cit s 2 has the potential to trigger clinical symptoms in orange allergy while at the same time cross-reactivity not correlated with clinical food allergy is also common and may reflect low affinity binding to orange profilin of IgE from subjects sensitized to profilins from various pollen sources.…”

Section: Discussionmentioning

confidence: 99%

Germin‐like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits

Crespo

Retzek

Foetisch

et al. 2006

Molecular Nutrition Food Res

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Oranges are clinically relevant allergenic foods. To date, orange allergens have not been characterized in detail. The study is aimed at analyzing the sensitization profile in orange-sensitized subjects with and without clinical allergy, and to identify orange allergens. Fifty-six sensitized subjects with selfreported reactions to orange were grouped into reactors (anaphylaxis or multiple episodes of immediate reactions and/or positive challenge tests) and non-reactors (negative open food challenge tests). Allergens were characterized by IgE immunoblotting, N-terminal sequencing, IgE-inhibition assays, and mediator release assays were performed to determine the allergenic potency of orange profilin. Of 56 subjects, 23 were classified as orange allergic showing mainly an oral allergy syndrome. Of 23 subjects classified as orange allergic, 22 were sensitized to profilin, Cit s 2. In patients with monosensitization to profilin in vitro histamine releases up to 75% from basophils were induced using orange extract and purified plant profilins. Of the allergic patients 78% were sensitized to germinlike protein, Cit s 1. Both allergens showed retained IgE reactivity in heat-processed orange juice. Interestingly, subjects with and without clinical allergy showed a comparable sensitization profile. Profilin and germin-like proteins are major orange allergens. The potential clinical relevance of orange profilin was indicated by its strong capacity to release histamine from basophils. However, a predominant sensitization to both allergens in subjects without symptoms also indicates a high frequency of clinically insignificant sensitization.

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“…RANTES and TGF-␤1 in culture supernatants, serum, and SF were measured by sandwich ELISA, as described [20]. Briefly, a 4 g/ml solution of monoclonal antibodies to human RANTES and TGF-␤1 (R&D Systems) was added to a 96-well plate (Nunc, Roskilde, Denmark) and incubated overnight at 4 • C. After incubating the plate with blocking solution (phosphate buffered saline (PBS) containing 1% bovine serum albumin and 0.05% Tween 20) for 2 h at room temperature, test samples and the standard recombinant cytokines (R&D Systems) were added, and the plate was incubated at room temperature for 2 h. After washing four times with PBS containing Tween 20, 500 ng/ml of biotinylated monoclonal antibodies to human RANTES and TGF-␤1 (R&D Systems) was added, and the reactions were allowed to proceed for 2 h at room temperature.…”

Section: Enzyme-linked Immunosorbent Assay (Elisa) Of Rantes and Tgf-β1mentioning

confidence: 99%

Transforming growth factor beta 1(TGF-β1) down-regulates TNFα-induced RANTES production in rheumatoid synovial fibroblasts through NF-κB-mediated transcriptional repression

et al. 2006

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Quantification of profilins by a monoclonal antibody-based sandwich ELISA

Cited by 26 publications

References 31 publications

Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA

Germin‐like protein Cit s 1 and profilin Cit s 2 are major allergens in orange (Citrus sinensis) fruits

Transforming growth factor beta 1(TGF-β1) down-regulates TNFα-induced RANTES production in rheumatoid synovial fibroblasts through NF-κB-mediated transcriptional repression

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