Simulated gastric fluid assay for estimating the digestibility of newly expressed proteins in GE crops: Missteps in development and interpretation.

Digestive stability of a food protein in simulated gastric fluid (SGF) continues to be considered a risk factor for allergy, even though the current science does not support this belief. Methodological shortcomings of the adaption of the SGF assay for use with purified proteins has been cited as a reason to discount results that do not conform to this belief. Missteps in conducting and interpreting the results of SGF assays are reviewed here. However, these methodological shortcomings do not invalidate the conclusion that allergenic proteins are not systematically more stable to digestion than non-allergens. The growing evidence for the dual allergen exposure hypothesis, whereby sensitization to food allergens is primarily caused by dermal and inhalation exposure to food dust, and tolerization against food allergy is primarily induced by gut exposure in food, likely explains why the digestive stability of a protein is not a risk factor for allergenicity.

Slow alignment of GMO allergenicity regulations with science on protein digestibility.

The current science on food allergy supports the dual allergen exposure hypothesis where sensitization to allergenic proteins is favored by dermal and inhalation exposure, and tolerization against allergy is favored by exposure in the gut. This hypothesis is bolstered by the epidemiological evidence showing that regions where children are exposed early in life to allergenic foods have lower rates of allergy. This led medical experts to replace the previous recommendation to exclude commonly allergenic foods from the diets of young children with the current recommendation that such foods be introduced to children early in life. Past beliefs that lowering gut exposure would reduce the likelihood that a protein would be allergenic led regulators and risk assessors to consider digestively stable proteins to be of greater allergenic risk. This resulted in international guidance and government regulations for newly expressed proteins in genetically engineered crops that aligned with this belief. Despite empirical results showing that allergens are no more digestively stable than non-allergens, and that gut exposure favors tolerization over sensitization, regulations have not come into alignment with the current science prompting developers to continue to engineer proteins for increased digestibility. In some rare cases, this could potentially increase sensitization risk.

Evidence runs contrary to digestive stability predicting protein allergenicity.

A dogma has persisted for over two decades that food allergens are more stable to digestion compared with non-allergenic proteins. This belief has become enshrined in regulations designed to assess the allergenic risk of novel food proteins. While the empirical evidence accumulated over the last 20+ years has largely failed to confirm a correlation between digestive stability and the allergenic status of proteins, even those who accept this finding often assert that this shortfall is the result of faulty assay design rather than lack of causality. Here, we outline why digestive stability may not in fact correlate with allergenic potential.

Characteristics and safety assessment of intractable proteins in genetically modified crops.

Genetically modified (GM) crops may contain newly expressed proteins that are described as "intractable". Safety assessment of these proteins may require some adaptations to the current assessment procedures. Intractable proteins are defined here as those proteins with properties that make it extremely difficult or impossible with current methods to express in heterologous systems; isolate, purify, or concentrate; quantify (due to low levels); demonstrate biological activity; or prove equivalency with plant proteins. Five classes of intractable proteins are discussed here: (1) membrane proteins, (2) signaling proteins, (3) transcription factors, (4) N-glycosylated proteins, and (5) resistance proteins (R-proteins, plant pathogen recognition proteins that activate innate immune responses). While the basic tiered weight-of-evidence approach for assessing the safety of GM crops proposed by the International Life Sciences Institute (ILSI) in 2008 is applicable to intractable proteins, new or modified methods may be required. For example, the first two steps in Tier I (hazard identification) analysis, gathering of applicable history of safe use (HOSU) information and bioinformatics analysis, do not require protein isolation. The extremely low level of expression of most intractable proteins should be taken into account while assessing safety of the intractable protein in GM crops. If Tier II (hazard characterization) analyses requiring animal feeding are judged to be necessary, alternatives to feeding high doses of pure protein may be needed. These alternatives are discussed here.