title: Two-dimensional electrophoresis and western-blotting analyses with anti Ara h 3 basic subunit IgG evidence the cross-reacting polypeptides of Arachis hypogaea, Glycine max, and Lupinus albus seed proteomes.
abstract: The allergenicity of seed storage proteins, the major components of edible legume seeds, may cause serious reactions in both children and adult population. Updated methodologies for evaluation of the activity of these proteins are needed. In this paper we used two-dimensional (2D) electrophoretic techniques to investigate the immuno-cross-reactivities of anti Ara h 3 basic subunit IgG to the seed proteomes of three legume species, namely, peanut, soybean, and lupin. The seed proteins, extracted with two different procedures, were separated by 2D electrophoresis, and the electrophoretic maps were analyzed by Western blot. In peanut proteome the antibodies strongly reacted with the 23 kDa polypeptides, corresponding to Ara h 3 basic isoforms, the antigen they were raised to, and three unidentified acidic polypeptides near 45 kDa. Remarkable cross-reactivities with lupin and soybean Ara h 3 homologous polypeptides and nonrelated proteins, namely, lupin conglutin gamma and soybean Bg7S, were detected. Therefore, these proteins may be regarded as new putative allergens. The present findings show the potentiality of 2D electrophoresis in the identification of food allergens and open the way to the traceability of the new cross-reacting proteins in the food chain.
authors.affiliation: Department of Agrifood Molecular Sciences, University of Milan, via Celoria 2, 20133 Milan, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Magni
authors.forename: Cinzia
authors.initials: C
authors.lastname: Ballabio
authors.forename: Patrizia
authors.initials: P
authors.lastname: Restani
authors.forename: Elena
authors.initials: E
authors.lastname: Sironi
authors.forename: Alessio
authors.initials: A
authors.lastname: Scarafoni
authors.forename: Claudio
authors.initials: C
authors.lastname: Poiesi
authors.forename: Marcello
authors.initials: M
authors.lastname: Duranti
mesh_terms: D000485:Allergens / Q000032:analysis / Q000276:immunology*; D000595:Amino Acid Sequence; D000918:Antibody Specificity; D052179:Antigens, Plant; D010367:Arachis / Q000737:chemistry* / Q000276:immunology; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D007073:Immunoglobulin E; D031297:Lupinus / Q000737:chemistry* / Q000276:immunology; D008969:Molecular Sequence Data; D020543:Proteome / Q000737:chemistry / Q000276:immunology*; D055314:Seed Storage Proteins; D012639:Seeds / Q000737:chemistry / Q000276:immunology; D013025:Glycine max / Q000737:chemistry* / Q000276:immunology
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D020543:Proteome; D055314:Seed Storage Proteins; C118207:allergen Ara h3; D007073:Immunoglobulin E