Localization of an IgE epitope of glycinin A2 peptide chain.

Abstract

One of the main allergens in soybeans is glycinin, which seriously impacts the normal lives of allergic people. Previous studies have confirmed that thermal processing and thermal processing combined with ultrahigh-pressure processing could significantly reduce the antigenicity of glycinin. The dominant antigen region of acidic peptide chain A2 of G2 subunit was located by phage display experiment. In this paper, overlapping peptides and alanine substitution techniques were used to explore the key amino acids that significantly affect the antigenicity of A2 peptide chain. The purity of peptide 1, peptide 2 and peptide 3 was identified by mass spectrometry and high-performance liquid chromatography, and the results showed that the purity of the synthesized overlapping peptide was more than 90%. SDS-PAGE showed that the peptide was successfully coupled with bovine serum albumin. The antigenicity of the coupling peptide was tested by ELISA and Dot-Blot, and the allergenicity was detected by reacting with the serum of patients with soybean globulin allergy. The results showed that peptide 3 has stronger antigenicity and sensitization. Alanine substitution technology allowed one to perform site-directed mutagenesis on peptide 3. Dot-Blot and ELISA tests showed that D259, E260, E261, Q263 and C266 may be the key amino acids that significantly affect the antigenicity of peptide 3. The research presented is of great significance for correctly guiding the production of safe food and preventing the occurrence of food allergic diseases. © 2023 Society of Chemical Industry.