Impact of high-pressure steam on Ara h 3 antigenicity and the location of the sensitization disruption region.

The globulin fraction of Arachis hypogaea is allergenic and the Ara h 3 protein in this globulin fraction is an important allergen. In this investigation, high-pressure steam treatment of peanut allergen Ara h 3 achieved a maximum antigenicity reduction of 76.16% under optimized conditions (10 mg/mL, 125 °C, 30 min). Subsequently, through multiple rounds of segmented expression and screening using phage display technology, combined with validation using high-pressure steam-treated antibodies and ELISA with serum from allergic patients, we ultimately identified the advantageous antigenic region Ara h 3-B-II-1 (173FNLAGNTEQEFLRYQQQSRQSRRRSLPYSPYS204). This area was split into three peptides, and Dot-blot and ELISA experiments demonstrated that Peptide 3 included linear sites. Substitution tests with alanine revealed that R191, Q192, R194, R196, L198, and P199 are important binding sites for IgE and IgG. Our findings provide theoretical guidelines for creating precise and effective peanut desensitization strategies.