title: Epitope mapping and identification of amino acids critical for mouse IgG-binding to linear epitopes on Gly m Bd 28K.
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.
authors.affiliation: a College of Food Science and Technology , Henan University of Technology , Zhengzhou , People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xi
authors.affiliation: a College of Food Science and Technology , Henan University of Technology , Zhengzhou , People's Republic of China.
authors.forename: Huili
authors.initials: H
authors.lastname: Yan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D019295:Computational Biology; D018604:Epitope Mapping; D006023:Glycoproteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D007074:Immunoglobulin G / Q000276:immunology*; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D030262:Soybean Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism
chemical_list: D052179:Antigens, Plant; C107967:Gly m Bd 28K allergen, Glycine max; D006023:Glycoproteins; D007074:Immunoglobulin G; D010455:Peptides; D030262:Soybean Proteins
keywords: Gly m Bd 28K; IgG-binding epitope; bioinformatics tool; monoclonal antibody; overlapping peptide