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Identification of allergenic epitopes destroyed by two processing technologies of glycinin A2 from soybean.

BACKGROUND
Glycinin is one of the most highly allergenic proteins in soybeans, and G2 is one of the five allergenic subunits of glycinin. Compared with the alkaline chain, the acidic chain A2 of the G2 subunit has strong allergenicity. However, the precise epitopes of A2 and the epitopes destroyed during processing are still unknown.


RESULTS
In this study, preparation of two specific antibodies damaged by processing, and phage display techniques were applied to locate the antigenic epitopes of glycinin A2 polypeptide chains disrupted by two processing techniques (thermal processing and ultra-high pressure combined thermal processing). Using bioinformatics methods to predict the possible epitopes of the A2 chain. A2 chain and its overlapping segments were introduced into T7 phages and expressed on phage shell by phage display. An indirect enzyme-linked immunosorbent assay (iELISA) was used to screen for antigenic epitopes that had been disrupted by the two processing technologies. The results shown that the dominant antigenic region disrupted by processing was located mainly in the A2-3-B fragment. The reacting experiment with the serum of allergic patients showed that the A2-3-B fragment protein was not only an antigenic region, but also an allergenic region. The two processing technologies destroyed the allergenic epitopes of A2 chain, thereby reducing the allergenicity of protein, and the amino acids where the dominant allergenic region disrupted by processing was located were 233 AIVTVKGGLRVTAPAMRKPQQEEDDDDEEEQPQCVE268 .


CONCLUSION
Precise epitopes of the acidic chain A2 in glycinin were identified, and epitopes destroyed in two common processing methods were also obtained. The application products of rapid detection of de-allergenicity effect of processed food can be developed according to the location of processed destruction allergenic region, which is of great significance in preventing the occurrence of soybean allergenic diseases. This article is protected by copyright. All rights reserved.