Epitope mapping and identification of amino acids critical for mouse IgG-binding to linear epitopes on Gly m Bd 28K. Issue 10 (2nd October 2016)

• Record Type: Journal Article
• Authors: Xi, Jun; Yan, Huili
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 80:Issue 10(2016)
• Page Start: 1973
• Page End: 1979
• Publication Date: 2016-10-02
• Subjects: Gly m Bd 28K -- IgG-binding epitope -- overlapping peptide -- monoclonal antibody -- bioinformatics tool; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2016.1165604
• Languages: English
• ISSNs: 0916-8451

Abstract

Abstract: Gly m Bd 28K is one of the major allergens in soybeans, but there is limited information on its IgG-binding epitopes. Thirty-four overlapping peptides that covered the entire sequence of Gly m Bd 28K were synthesized, and 3 monoclonal antibodies against Gly m Bd 28K were utilized to identify the IgG-binding regions of Gly m Bd 28K. Three dominant peptides corresponding to 28 GDKKSPKSLFLMSNS 42 (G28-S42), 56 LKSHGGRIFYRHMHI 70 (L56-I70), and 154 ETFQSFYIGGGANSH 168 (E154-H168) were recognized. L56-I70 is the most important epitope, and a competitive ELISA indicated that it could inhibit the binding of monoclonal antibody to Gly m Bd 28K protein. Alanine scanning of L56-I70 documented that F64, Y65, and R66 were the critical amino acids of this epitope. Two bioinformatics tools, ABCpred and BepiPred, were used to predict the epitopes of Gly m Bd 28K, and the predictions were compared with the epitopes that we had located by monoclonal antibodies. Graphical abstract: : Epitope mapping and identification of amino acids critical for mouse IgG-binding to linear epitopes on Gly m Bd 28K.