Investigation of incurred single- and multi-component model food matrices for determination of food proteins triggering allergy and coeliac disease

Török, Kitti; Horvath, V. A. P.; Horváth, Ágnes; Hajas, Lívia; Bugyi, Zsuzsanna; Tömösközi, Sándor

doi:10.1007/s00217-014-2289-6

Cited by 16 publications

(7 citation statements)

References 32 publications

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“…When analyzing processed foods, such as meat products, polymerase chain reaction (PCR) techniques can be very useful alternatives to immunochemical methods [6]. Until now, the detection of milk allergens in foodstuffs has been mostly performed by protein analysis, relying on mass spectrometry [7][8][9][10][11] and ELISA [12][13][14][15]. However, the application of PCR-based methods to detect and quantify milk as an allergenic food is still limited.…”

Section: Introductionmentioning

confidence: 99%

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

Villa

Costa

2019

Biomolecules

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Milk ingredients are often included in a wide range of meat products, such as cooked hams and sausages, to improve technological characteristics. However, milk proteins are also important food allergens. The aim of this study was the development of a highly sensitive and specific real-time PCR system targeting the 12S rRNA gene of Bos domesticus for the detection and quantification of milk as an allergenic ingredient in processed meat products. The method was able to achieve an absolute limit of detection (LOD) of 6 fg of milk DNA. Using a normalized approach (∆Ct method) for the detection of milk protein concentrate (MPC), it was possible to obtain sensitivities down to 0.01% (w/w) of MPC in model hams (raw and cooked) and autoclaved sausages, and 0.005% in raw sausage mixtures. The developed systems generally presented acceptable PCR performance parameters, being successfully validated with blind samples, applied to commercial samples, and further compared with an immunochemical assay. Trace amounts of milk material were quantified in two out of 13 samples, but the results mostly infer the excessive practice of the precautionary labeling.

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Section: Introductionmentioning

confidence: 99%

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

Villa

Costa

2019

Biomolecules

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“…For effi cient extraction of prolamins from processed products, extraction solvents containing reducing and disaggregating agents were developed (GARCÍA et al, 2005;GESSENDORFER et al, 2010;MENA et al, 2012). Matrix effects caused by different macro and micro components of foods may also change the extraction yield and immunochemical reactivity of gluten proteins through the formation of complexes or resistant aggregates (SCHERF & POMS, 2016;TÖRÖK et al, 2014TÖRÖK et al, , 2015.…”

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confidence: 99%

ELISA response and gliadin composition of different wheat cultivars grown in multiple harvest years

et al. 2017

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In special dietary products for people intolerant to gluten, gluten content is not supposed to exceed the regulatory thresholds. Enzyme-linked immunosorbent assays (ELISAs) are routinely used to quantitate gluten in these products. They measure gliadin/gluten with high specifi city and sensitivity, but they have some limitations. Quantitative and qualitative variability of the target proteins among wheat cultivars is a factor that may cause inaccurate results. One of the aims of this work was to characterize the protein composition of fi ve wheat cultivars grown in multiple harvest years and their blends by reversed-phase high-performance liquid chromatography (RP-HPLC). The gliadin/gluten content of these wheat fl ours was also analysed with two commercial ELISA kits. The effect of differences in protein profi les between the fl ours from an individual cultivar and the blend of fi ve cultivars, harvest years, as well as processing procedures (dough forming and baking) on the results of two ELISA kits was investigated and their relative magnitude was determined. Among the factors investigated, the differences between fl ours had the greatest impact on gliadin recoveries.Keywords: wheat fl our, protein composition, gluten-free, gliadin/gluten, incurred matrix, ELISAThe ingestion of food from gluten-containing grains can trigger a number of harmful reactions in susceptible individuals, e.g. celiac disease, wheat allergy, and non-celiac gluten sensitivity. Gluten (especially its prolamin fraction) from wheat, rye, and barley is the precipitating factor for celiac disease . Patients must eliminate these grains from their diet, because currently this is the only effective strategy to avoid the symptoms (SABATINO et al., 2013). Two thresholds are defi ned: the terms 'gluten-free' and 'very low gluten' mean that the gluten content in these products is below 20 and 100 mg kg -1, respectively (COMMISSION OF THE EUROPEAN COMMUNITIES, 2009). Reliable analytical methods are needed to assess the gluten content of these products and to enforce regulations. Currently, immunochemical assays (enzyme-linked immunosorbent assays (ELISAs) and lateral fl ow devices (LFDs)) are the preferred methods for determining such low amounts of protein. However, gluten quantitation with ELISA is accompanied by numerous problems, because gluten is a complex mixture of heterogeneous proteins with different solubility, composition, and immunoreactivity (DIAZ- .Most commercial gluten immunoassays are based on determining prolamins, which are soluble in aqueous alcohols (SCHERF & POMS, 2016). Although the primary structure of prolamins appears to be relatively stable, food processing might induce post-translational * To whom correspondence should be addressed. Phone: +36-1-463-1419; fax: +36-1-463-3855; e-mail: tomoskozi@mail.bme.huThis is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 License, which permits unrestricted use, distribution, and reproduction in any medium for non-commercial purposes, provid...

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“…A multicomponent model product was also produced, which contained 2-2% of each allergen component. The production procedure was previously published (TÖRÖK et al, 2014b). Briefl y, in the fi rst step powder mixtures were made with the mixing of the dry components of the recipe and the allergen source(s).…”

Section: Methodsmentioning

confidence: 99%

Identification of key effects causing weak performance of allergen analysis in processed food matrices

et al. 2016

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The weaker performance of generally used analytical methods for allergen analysis in processed foods can be connected to protein denaturation. To understand the nature of protein denaturation processes, experimental but realistic model matrices (corn starch based mixture, hydrated dough, and heat treated cookies) were developed that contain a defi ned amount of milk, egg, soy, and wheat proteins individually or in combination. The protein subunit composition was investigated in every processing phase, i.e. after mixing, dough formation, and baking. SDS-PAGE measurements were carried out to monitor the protein distribution of sample food matrices in non-reducing and reducing gels. The results clearly show that the highly decreased protein solubility is caused by denaturation, aggregation, or complex formation, which are the most signifi cant factors in poorer analytical performances. Solubility can only partly be improved with the application of reducing agents or surfactants, and the rate of improvement is depending on the proteins and the matrices.

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Investigation of incurred single- and multi-component model food matrices for determination of food proteins triggering allergy and coeliac disease

Cited by 16 publications

References 32 publications

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

Detection and Quantification of Milk Ingredients as Hidden Allergens in Meat Products by a Novel Specific Real-Time PCR Method

ELISA response and gliadin composition of different wheat cultivars grown in multiple harvest years

Identification of key effects causing weak performance of allergen analysis in processed food matrices

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