Stepwise hydrolysis reduces allergenicity of α-lactalbumin through epitope removal and structural modulation.

Hydrolysis is an effective strategy to reduce the allergenicity of α-lactalbumin (ALA). In this study, the low-allergenicity hydrolysate (FA) was prepared by using the stepwise hydrolysis technology, and its allergenicity was evaluated from multiple perspectives. The results showed that compared to one-step hydrolysates, FA not only had higher hydrolysis efficiency and further eliminated linear allergenic epitopes, but also significantly reduced antigenicity. The increased degree of hydrolysis leads to reduced IgE binding and subsequent inhibition of mast cell degranulation, thereby better maintaining the Th1/Th2 immune balance. In conclusion, stepwise hydrolysis is an effective strategy for reducing ALA allergenicity. This study provides preliminary mechanistic evidence and a methodological reference for the further development and validation of hypoallergenic dairy ingredients.