Development of a low allergenic product for patients with milk allergy and assessment of its specific IgE reactivity

Abstract

AbstractBackgroundMilk oral immunotherapy is the riskiest and most unpredictable form of oral immunotherapy. We aimed to produce a low allergenic product than conventional once baked‐cake/muffin, to develop indirect in‐house ELISA to check the tolerance status with milk products and evaluate IgE reactivity of patients' sera via western blotting (WB) and indirect in‐house ELISA.MethodA low allergenic product named biscotti‐twice baked‐cake was developed, and the total protein concentration was determined. The protein content was studied by SDS‐PAGE and proteomics. Milk‐specific IgE (sIgE) binding assays were performed by WB and indirect in‐house ELISA by using patients' sera.ResultsCasein band intensity was observed to be lower in the biscotti‐twice baked‐cake than in the once baked‐cake (p = .014). Proteomics analysis and αS1‐casein measurement showed that the lowest intensity of casein was found in biscotti. The low binding capacity of milk sIgE to biscotti compared with once baked‐cake was shown by WB (p = .0012) and by indirect in‐house ELISA (p = .0001). In the ROC analysis, the area under the curve (AUC) of the in‐house ELISA IgE was comparable with Uni‐CAP milk and casein sIgE. The AUC of the in‐house ELISA IgE for cake (0.96) and biscotti (1) was slightly better than Uni‐CAP milk sIgE (0.94; 0.97) and casein sIgE (0.96; 0.97), respectively.ConclusionThe low allergenicity of the newly developed low allergenic product “biscotti‐twice baked‐cake” has been demonstrated by in vitro experiments. Biscotti could be a safe treatment option than once baked‐cake/muffin in patients who are reactive to once baked‐milk products.