Identification of antigenic sites destructed by high hydrostatic pressure (HHP) of the β subunit of β-conglycinin

• Published in: International journal of biological macromolecules Vol. 141; pp. 1287 - 1292
• Main Authors: Xi, Jun, He, MengXue, Pi, JiangYi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2019
• Subjects: Amino Acid Sequence; Antigens, Plant - chemistry; Antigens, Plant - genetics; Antigens, Plant - immunology; Bacteriophages - genetics; DNA, Recombinant - genetics; Epitopes - genetics; Genetic Engineering; Globulins - chemistry; Globulins - genetics; Globulins - immunology; HHP destructive antigenic sites; Hydrostatic Pressure; Models, Molecular; Protein Conformation; Protein Subunits - chemistry; Protein Subunits - genetics; Protein Subunits - immunology; Seed Storage Proteins - chemistry; Seed Storage Proteins - genetics; Seed Storage Proteins - immunology; Soybean Proteins - chemistry; Soybean Proteins - genetics; Soybean Proteins - immunology; T7 phage; β subunit; β subunit; T7 phage; HHP destructive antigenic sites
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2019.09.042

β-conglycinin is one of the most allergenic proteins, and its constituent subunits α′, α, and β are all potential allergens to humans. In the present study, we concentrated on the destructed antigenic sites of β subunit of β-conglycinin after high hydrostatic pressure (HHP) treatment. In this paper, the overlapping gene fragments of the β subunit of β-conglycinin were amplified by polymerase chain reaction (PCR) and cloned into T7 phage vectors. After being packaged in vitro, the recombinant T7 phage was constructed, and the overlapping fragments of the β subunit were displayed on the phage surface. The recombinant phages that expressed the overlapping fragments of the β subunit were used to react with specific antiserum by indirect ELISA to identify the HHP destructed antigenic sites. After three rounds of expression and identification, we used synthetic peptide technology to identify that the obtained fragment was a conformational epitope. We further confirmed that HHP treatment changed the conformational structure of β-conglycinin, which reduced the antigenicity of the protein.