Protein Digestibility and Relevance to Allergenicity

Current allergy and asthma reports, 2004

Food allergens are almost always proteins, but not all food proteins are allergens. This one statement sums up the purpose of this article, defining the difference between an innocuous food protein and a food allergen. The simplest answer is that a food allergen has the ability to first elicit an IgE response, and then, on subsequent exposures, to elicit a clinical response to the same or similar protein. However, this simplistic answer avoids the more complex issues of defining the biochemical characteristics that allow a food protein to survive the extremes of food processing, escape the digestive enzymes of the human gastrointestinal tract, and interact with the immune system. More than 700 allergen sequences have been identified from food and nonfood sources. However, despite increasing knowledge of the structure and amino acid sequences of the identified allergens, only a few biochemical characteristics can be associated with food allergens. Food allergen characteristics, inclu...

Food and Chemical Toxicology, 2019

The current allergenicity assessment of novel proteins is based on the EFSA GMO guidance. Recently, EFSA launched a new guidance document on allergenicity assessment of GM plants (2017). This document describes, amongst other topics, the new scientific and regulatory developments on in vitro protein digestibility tests. The EFSA GMO Panel stated that for in vitro protein digestibility tests, additional investigations are needed before any additional recommendation in the form of guidance can be provided. To this end, an interim phase is considered necessary to evaluate the revisions to the in vitro gastrointestinal digestion test, proposed by EFSA. This prompted the establishment of a joint workshop through two COST Action networks: COST Action ImpARAS and COST Acton INFOGEST. In 2017, a workshop was organised to discuss the relevance of digestion in allergenicity risk assessment and how to potentially improve the current methods and readouts. The outcome of the workshop is that there is no rationale for a clear readout that is predictive for allergenicity and we suggest to omit the digestion test from the allergenicity assessment strategy for now, and put an effort into filling the knowledge gaps as summarized in this paper first.

Journal of Agricultural and Food Chemistry, 2013

Current models of digestibility utilize pepsin stability to assess the safety of allergenic versus nonallergenic food proteins. Dietary protein digestion in vivo, however, requires acid denaturation and protease cleavage by pepsin, trypsin, and/or chymotrypsin. The ability of this approach to identify food protein stability in the mammalian gut may be limited. We determined the temporal stability and immunoreactivity of almond, pine nut, and peanut allergenic proteins under simulated physiologic gastric and intestinal digestive conditions in vitro. Gel electrophoresis and immunoblot analyses were used to determine protein stability and immunoreactivity, respectively. Peanut, almond, and pine nut proteins were pepsin-and pancreatin-stable and immunoreactive for up to 1 h after initiation of digestion. Moreover, successive acid denaturation and pepsin and pancreatin cleavage were necessary to hydrolyze these allergenic proteins and reduce their IgG-and IgE-binding capacity, which suggests that digestibility models must be improved for more accurate safety assessment of food allergens.

Environmental Health Perspectives, 2002

Because of the public concern surrounding the issue of the safety of genetically modified organisms, it is critical to have appropriate methodologies to aid investigators in identifying potential hazards associated with consumption of foods produced with these materials. A recent panel of experts convened by the Food and Agriculture Organization and World Health Organization suggested there is scientific evidence that using data from animal studies will contribute important information regarding the allergenicity of foods derived from biotechnology. This view has given further impetus to the development of suitable animal models for allergenicity assessment. This article is a review of what has been achieved and what still has to be accomplished regarding several different animal models. Progress made in the design and evaluation of models in the rat, the mouse, the dog and in swine is reviewed and discussed.

Toxicological Sciences, 1999

The ability of exogenous proteins to cause respiratory and gastrointestinal allergy, and sometimes systemic anaphylactic reactions, is well known. What is not clear however, are the properties that confer on proteins the ability to induce allergic sensitization. With an expansion in the use of enzymes for industrial applications and consumer products, and a substantial and growing investment in the development of transgenic crop plants that express novel proteins introduced from other sources, the issue of protein allergenicity has assumed considerable toxicological significance. There is a need now for methods that will allow the accurate identification and characterization of potential protein allergens and for estimation of relative potency as a first step towards risk assessment. To address some of these issues, and to review progress that has been made in the toxicological investigation of respiratory and gastrointestinal allergy induced by proteins, a workshop, entitled the Toxicology of Protein Allergenicity: Prediction and Characterization, was convened at the 37 th Annual Conference of the Society of Toxicology in Seattle, Washington (1998). The subject of protein allergenicity is considered here in the context of presentations made at that workshop.

PLOS One, 2009

Background: Predicting the allergenicity of proteins is challenging. We considered the possibility that the properties of the intact protein that may alter the likelihood of being taken up by antigen presenting cells, may be useful adjuncts in predicting allergens and non-allergens in silico. It has been shown that negatively charged acidic proteins are preferentially processed by dendritic cells.

Clinical and translational allergy, 2018

Susceptibility to pepsin digestion of candidate transgene products is regarded an important parameter in the weight-of-evidence approach for allergenicity risk assessment of genetically modified crops. It has been argued that protocols used for this assessment should better reflect physiological conditions encountered in representative food consumption scenarios. To evaluate whether inclusion of more physiological conditions, such as sub-optimal and lower pepsin concentrations, in combination with pancreatin digestion, improved the performance of digestibility protocols used in characterization of protein stability. Four pairs of established allergens and their related non/weakly-allergenic counterparts (seed albumins, muscle tropomyosins, plant lipid transfer proteins [LTP] and collagens) plus fish parvalbumin, were subjected to nine combinations of pH (1.2-2.5-4.0) and pepsin-to-protein ratio (PPR: 10-1-0.1 U/µg) for pepsin digestion, followed by pancreatin digestion in the presen...

Food and Chemical Toxicology, 2008

a b s t r a c t in Nice, France, to review and discuss existing and emerging methods and techniques for improving the current weightof-evidence approach for evaluating the potential allergenicity of novel proteins. The workshop included over 40 international experts from government, industry, and academia. Their expertise represented a range of disciplines including immunology, chemistry, molecular biology, bioinformatics, and toxicology. Among participants, there was consensus that (1) current bioinformatic approaches are highly conservative; (2) advances in bioinformatics using structural comparisons of proteins may be helpful as the availability of structural data increases; (3) proteomics may prove useful for monitoring the natural variability in a plant's proteome and assessing the impact of biotechnology transformations on endogenous levels of allergens, but only when analytical techniques have been standardized and additional data are available on the natural variation of protein expression in non-transgenic bred plants; (4) basophil response assays are promising techniques, but need additional evaluation around specificity, sensitivity, and reproducibility; (5) additional research is required to develop and validate an animal model for the purpose of predicting protein allergenicity.

Proteins in food …, 2004

In order to take a systematic approach as to how allergens may be removed from foods using approaches such as raw material selection or physical processing it is important to have a clear understanding of what constitutes an allergen. Taken in its strictest sense an allergen is defined as a macromolecule to which immunoglobulin E (IgE) binds. IgE responses are generated as part of the normal immune reaction to parasitic infections, but for reasons that are only partly understood can also be generated following exposure to environmental agents, such as pollen, dusts, and foods. On a subsequent encounter the allergen is able to cross-link surface bound IgE molecules located on basophile or mast cells, causing them to release preformed inflammatory mediators, including histamine. It is these mediators which actually cause the symptoms manifested in an allergic episode which usually occurs quite rapidly following exposure and is consequently classified as a Type I hypersensitivity reaction. Such reactions are distinct from food intolerances, which are reproducible, sometimes non-immune-mediated, reactions whose symptoms can take days to manifest themselves. An example of such a condition is the gluten intolerance syndrome, Coeliac disease, and whilst the gluten proteins which trigger it are not IgE-binding proteins, some workers refer to them as allergens, and they are treated as such in the WHO Codex Alimentarius Commission Labelling regulations. Almost all allergens are proteinaceous in nature, as other polymers, such as polysaccharides, usually generate only poor antibody responses. There is

Allergy, 2019

Significant efforts are necessary to introduce new dietary protein sources to feed a growing world population while maintaining food supply chain sustainability. Such a sustainable protein transition includes the use of highly modified proteins from side streams or the introduction of new protein sources that may lead to increased clinically relevant allergic sensitization. With food allergy being a major health problem of increasing concern, understanding the potential allergenicity of new or modified proteins is crucial to ensure public health protection. The best predictive risk assessment methods currently relied on are in vivo models, making the choice of endpoint parameters a key element in evaluating the sensitizing capacity of novel proteins. Here, we provide a comprehensive overview of the most frequently used in vivo and ex vivo endpoints in murine food allergy models, addressing their strengths and limitations for assessing sensitization risks. For optimal labto-lab reproducibility and reliable use of predictive tests for protein risk assessment, it is important that researchers maintain and apply the same relevant parameters and Accepted Article This article is protected by copyright. All rights reserved. procedures. Thus, there is an urgent need for a consensus on key food allergy parameters to be applied in future food allergy research in synergy between both knowledge institutes and clinicians.