Protein Ingredients Control in Gluten Free Products Using SDS-PAGE, Developed Competitive Enzyme Immunoassays and Commercial ELISA Kits.

Some protein ingredients declared in the label of gluten free products are allergenic proteins (milk, soy and egg).The proper identification of these proteins in food products is important for consumers who have food allergies. The aim of this study was to control the presence of protein ingredients declared in the label of gluten free products. Samples were analyzed with sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), using an extractive solution of total proteins (Tris-ClH buffer 0.0625 M with 3% sodium dodecylsulfate and 2% 2-mercaptoethanol; pH: 6.8) and a selective solvent for the extraction of caseins (isopropanol 55° + 2-mercaptoethanol / ISO + ME). Developed Competitive enzyme immunoassays were used for the detection / quantification of milk, soy and egg proteins in products elaborated with rice flour. Specific polyclonal rabbit antiserums against milk, soy and egg proteins were used as primary antibodies in these competitive enzyme immunoassays. Commercial ELISA kits from Neogen, R-Biopharm and Romer were used to verify the results. In some samples undeclared allergens were detected. Correct allergens labeling is very important for the safety of allergic consumers. In conclusion, it is possible to identify all the proteins ingredients in these gluten-free foods studied, using a combination of electrophoretic methods and immunochemical methods.Fil: Cellerino, Karina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cagnasso, Carolina Elisa. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Greco, Carola Beatriz. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Docena, Guillermo H.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Polenta, Gustavo Alberto. Instituto Nacional de Tecnología Agropecuaria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ferreyra, Marcela Verónica. Instituto Nacional de Tecnología Industrial; ArgentinaFil: Lopez, Laura Beatriz. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

A Randomized, Open-Label Trial of Hen's Egg Oral Immunotherapy : Efficacy and Humoral Immune Responses in 50 Children

BACKGROUND: Egg allergy is the second most common food allergy in children. Persistent food allergy increases the risk of anaphylaxis and reduces the quality of life. OBJECTIVE: To determine the efficacy of oral immunotherapy (OIT) with raw egg white powder and study its effects on humoral responses in children with persistent egg allergy. METHODS: Fifty children aged 6 to 17 years with egg allergy, diagnosed by double-blind, placebo-controlled food challenge, were randomized 3:2 to 8 months of OIT with a maintenance dose of 1 g of egg white protein or 6 months of avoidance after which the avoidance group crossed over to OIT. We examined changes in IgE, IgG4, and IgA concentrations to Gal d 1-4 during OIT compared with avoidance and assessed clinical reactivity at 8 and 18 months. RESULTS: After 8 months, 22 of 50 children (44%) on OIT and 1 of 21 (4.8%) on egg avoidance were desensitized to the target dose, 23 of 50 (46%) were partially desensitized (dosePeer reviewe

Tracking a Major Egg Allergen to Assess Commercial Food Label Compliance: Towards a Simple and Fast Immunosensing Device

An amperometric immunosensor was developed for the analysis of the major egg-white allergen ovotransferrin (Gal d 3) in commercial food products because the (accidental) intake, skin contact with, and/or inhalation of eggs can lead to severe disorders in allergic individuals. Employing a sandwich-type immunosensing strategy, screen-printed carbon electrodes (SPCE) were biomodified with anti-Gal d 3 (capture) antibodies, and the allergen’s detection was achieved with anti-Gal d 3 antibodies labelled with horseradish peroxidase (HRP). The 3,3′,5,5′-tetramethylbenzidine (TMB)/H2O2 reaction with HRP was used to obtain the electrochemical (amperometric) signal. An attractive assay time of 30 min and a remarkable analytical performance was achieved. The quantification range was established between 55 and 1000 ng·mL−1, with a limit of detection of 16 ng·mL−1. The developed method demonstrated good precision (Vx0 = 5.5%) and provided precise results (CV < 6%). The sensor also detected extremely low amounts (down to 0.010%) of egg. The analysis of seven raw and/or cooked egg and egg-white samples indicated that food processing influences the amount of allergen. Furthermore, to assure the compliance of product labelling with EU legislation, 25 commercial food ingredients/products were analysed. The accuracy of the results was confirmed through an ELISA assay. The stability of the ready-to-use sensing surface for 20 days allows a reduction of the reagents’ volumes and cost.This research received financial support from the European Union (FEDER funds through COMPETE POCI-01-0145-FEDER-030735) and National Funds (Fundação para a Ciência e a Tecnologia (FCT)) through the project PTDC/QUI-QAN/30735/2017—TracAllerSens—Electrochemical sensors for the detection and quantification of trace amounts of allergens in food products.info:eu-repo/semantics/publishedVersio

Study protocol of a phase 2, dual-centre, randomised, controlled trial evaluating the effectiveness of probiotic and egg oral immunotherapy at inducing desensitisation or sustained unresponsiveness (remission) in participants with egg allergy compared with placebo (probiotic egg allergen oral immunotherapy for treatment of egg allergy: PEAT study)

Introduction Egg allergy is the most common food allergy in children but recent studies have shown persistence or delayed resolution into adolescence. As there is currently no effective long-term treatment, definitive treatments that improve quality of life and prevent fatalities for food allergies are required. We have previously shown that a novel treatment comprising a combination of the probiotic Lactobacillus rhamnosus CGMCC 1.3724 with peanut oral immunotherapy (OIT) is highly effective at inducing sustained unresponsiveness, with benefit persisting to 4 years after treatment cessation in the majority of initial treatment responders. In this study, we plan to extend the probiotic food OIT platform to another allergen, namely egg. We describe the protocol for a phase 2, dual-centre, randomised, controlled trial evaluating the effectiveness of probiotic and egg OIT at inducing desensitisation or sustained unresponsiveness (remission) in participants with egg allergy compared with placebo. Methods and analysis 80 participants aged 5-30 years of age with current egg allergy confirmed by double-blind placebo-controlled food challenge at study screening will be recruited from Australia and Singapore. There are two intervention arms - probiotic and egg OIT (active) or placebo. Interventions are administered once daily for 18 months. The primary outcome is the proportion of participants who attain 8-week sustained unresponsiveness in the active group versus placebo group. Ethics and dissemination This study has been approved by the Human Research Ethics Committees at the Royal Children's Hospital (HREC 2019.082) and the National Healthcare Group Domain Specific Review Board (2019/00029). Results will be published in peer-reviewed journals and disseminated via presentations at international conferences. Trial registration number ACTRN12619000480189

Nuts \u27n\u27 guts: transport of food allergens across the intestinal epithelium

The increase in the incidence of food allergy is a growing problem for the western world. This review will focus on the findings from several macromolecular epithelial transport experiments and drug permeability studies to provide a recent comprehension of food allergen intestinal epithelial cell transport and the allergen-epithelial relationship. Specifically, this review will aim to answer whether allergens can permeate the intestinal barrier directly via intestinal epithelial cells, and whether this mode of transport affects downstream immune reactions. By improving our understanding of the interactions which take place during exposure of food allergens with the intestinal epithelium, we can begin to understand whether the epithelial barrier plays a major role in the allergic sensitization process rather than simply restricting the entry of allergens to the underlying lamina propria

Improved Sensitivity of Allergen Detection by Immunoaffinity LC-MS/MS Using Ovalbumin as a Case Study

Vitamin D deficiency due to, e.g., nutritional and life style reasons is a health concern that is gaining increasing attention over the last two decades. Vitamin D3, the most common isoform of vitamin D, is only available in food derived from animal sources. However, mushrooms and yeast are rich in ergosterol. This compound can be converted into vitamin D2 by UV-light, and therefore act as a precursor for vitamin D. Vitamin D2 from UV-irradiated mushrooms has become an alternative source of vitamin D, especially for persons pursuing a vegan diet. UV-irradiated baker’s yeast (Saccharomyces cerevisiae) for the production of fortified yeast-leavened bread and baked goods was approved as a Novel Food Ingredient in the European Union, according to Regulation (EC) No. 258/97. The Scientific Opinion provided by the European Food Safety Authority Panel on Dietetic Products, Nutrition, and Allergies has assessed this Novel Food Ingredient as safe under the intended nutritional use. However, recent findings on the formation of side products during UV-irradiation, e.g., the photoproducts tachysterol and lumisterol which are compounds with no adequate risk assessment performed, have only been marginally considered for this EFSA opinion. Furthermore, proceedings in analytics can provide additional insights, which might open up new perspectives, also regarding the bioavailability and potential health benefits of vitamin D-fortified mushrooms and yeast. Therefore, this review is intended to give an overview on the current status of UV irradiation in mushrooms and yeast in general and provide a detailed assessment on the potential health effects of UV-irradiated baker’s yeast