Statement on in vitro protein digestibility tests in allergenicity and protein safety assessment of genetically modified plants

Abstract

This statement supplements and updates the GMO Panel guidance document on allergenicity of genetically modified (GM) plants published in 2017. In that guidance document, the GMO Panel considered that additional investigations on in vitro protein digestibility were needed before providing any additional recommendations in the form of guidance to applicants. Thus, an interim phase was proposed to assess the utility of an enhanced in vitro digestion test, as compared to the classical pepsin resistance test. Historically, resistance to degradation by pepsin using the classical pepsin resistance test has been considered as additional information, in a weight‐of‐evidence approach, for the assessment of allergenicity and toxicity of newly expressed proteins in GM plants. However, more recent evidence does not support this test as a good predictor of allergenic potential for hazard. Furthermore, there is a need for more reliable systems to predict the fate of the proteins in the gastrointestinal tract and how they interact with the relevant human cells. Nevertheless, the classical pepsin resistance test can still provide some information on the physicochemical properties of novel proteins relating to their stability under acidic conditions. But other methods can be used to obtain data on protein's structural and/or functional integrity. It is acknowledged that the classical pepsin resistance test is embedded into international guidelines, e.g. Codex Alimentarius and Regulation (EU) No 503/2013. For future development, a deeper understanding of protein digestion in the gastrointestinal tract could enable the framing of more robust strategies for the safety assessment of proteins. Given the high complexity of the digestion and absorption process of dietary proteins, it is needed to clarify and identify the aspects that could be relevant to assess potential risks of allergenicity and toxicity of proteins. To this end, a series of research questions to be addressed are also formulated in this statement.

Scientific Opinion on development needs for the allergenicity and protein safety assessment of food and feed products derived from biotechnology

“…The GMO Panel considered the principles described on allergenicity in its guidance documents, statements, and scientific opinions (EFSA GMO Panel, 2010 , 2011 , 2017 , 2021 ), Regulation (EU) No 503/2013 and other relevant international guidelines (Codex Alimentarius, 2003–2009 ).…”

“…EFSA has been proactive in this respect and has already invested resources to advance the allergenicity prediction further. A series of EFSA procurements were undertaken, which resulted in several publications representing significant steps forward (Mills et al, 2013a,b; Mackie et al., 2019 ; Parenti et al, 2019 ; EFSA GMO Panel, 2017 , 2021 ). Likewise, EU‐funded research programmes, such as the ImpARAS Cost Action, EuroPrevall, iFAAM and AllerScreening projects, among others, also provide insights on the use and improvement of existing and suggested assessment tools in the field of allergenicity assessment of foods.…”

“…Subsequently, an Ad hoc Allergenicity working group of the GMO Panel was established to address to what extent the in vitro digestion test adds value to the allergenicity risk assessment of GM plants and the protein safety assessment in general, and consequently, published a statement entitled ‘ in vitro protein digestibility tests in allergenicity and protein safety assessment of genetically modified plants’ (EFSA GMO Panel, 2021 ).…”

Assessment of genetically modified oilseed rape 73496 for food and feed uses, under Regulation (EC) No 1829/2003 (application EFSA‐GMO‐NL‐2012‐109)

“…Protein stability is one of several relevant parameters to consider in the weight-of-evidence approach in protein safety (EFSA GMO Panel, 2010c, EFSA GMO Panel, 2011a, EFSA GMO Panel, 2017, EFSA GMO Panel, 2021. The term protein stability encompasses several properties such as thermal stability, pH-dependent stability, proteolytic stability and physical stability (e.g.…”

Workshop on allergenicity assessment – prediction

“…The topics addressed in such a document were IgE-and non-IgE-mediated adverse immune reactions to foods and risk assessment with an emphasis on celiac disease, the role of in vitro protein digestibility tests and endogenous allergenicity. A follow-up EFSA external report (EFSA, 2019a) and a related Statement of the EFSA GMO Panel on in vitro protein digestion was recently published (EFSA GMO Panel, 2021).…”