title: Protocol for simultaneous isolation of three important banana allergens.
abstract: Banana fruit (Musa acuminata) has become an important food allergen source in recent years. So far, 5 IgE reactive banana proteins have been identified, and the major allergens are: Mus a 2 (a class I chitinase, 31kDa), Mus a 4 (thaumatin-like protein, 21kDa), and Mus a 5 (β-1,3-glucanase, 33kDa). Due to variations in allergen expression levels, diagnostic reagents for food allergy can be improved by using individual allergen components instead of banana allergen extracts. The purpose of this study was to optimize the purification protocol of the three major allergens present in banana fruit: Mus a 2, Mus a 4 and Mus a 5. By employing a three-step purification protocol (a combination of anion-exchange, cation-exchange and reversed-phase chromatography) three important banana allergens were obtained in sufficient yield and high purity. Characterization of the purified proteins was performed by both biochemical (2-D PAGE, mass fingerprint and N-terminal sequencing) and immunochemical (immunoblot) methods. IgE reactivity to the purified allergens was tested by employing sera of five allergic patients. The purified allergens displayed higher sensitivity in IgE detection than the routinely used extracts. The three purified allergens are good candidates for reagents in component-based diagnosis of banana allergy.
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia.
authors.forename: Jasna
authors.initials: J
authors.lastname: Nikolic
authors.affiliation: Innovation Center of the Faculty of Chemistry, University of Belgrade, Serbia.
authors.forename: Ivan
authors.initials: I
authors.lastname: Mrkic
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia.
authors.forename: Milica
authors.initials: M
authors.lastname: Grozdanovic
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia.
authors.forename: Milica
authors.initials: M
authors.lastname: Popovic
authors.affiliation: Division of Clinical and Molecular Allergology, Research Center Borstel, Airway Research Center North (ARCN), Member of the German Center for Lung Research, Borstel, Germany.
authors.forename: Arnd
authors.initials: A
authors.lastname: Petersen
authors.affiliation: Division of Clinical and Molecular Allergology, Research Center Borstel, Airway Research Center North (ARCN), Member of the German Center for Lung Research, Borstel, Germany; Department of Dermatology and Allergology, University of Luebeck, Germany.
authors.forename: Uta
authors.initials: U
authors.lastname: Jappe
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia. Electronic address: mgavrov@chem.bg.ac.rs.
authors.forename: Marija
authors.initials: M
authors.lastname: Gavrovic-Jankulovic
mesh_terms: D000328:Adult; D000368:Aged; D000485:Allergens / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D052179:Antigens, Plant / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D002688:Chitinases / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D005260:Female; D005512:Food Hypersensitivity / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D005638:Fruit / Q000737:chemistry; D043326:Glucan 1,3-beta-Glucosidase / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D008297:Male; D008875:Middle Aged; D028521:Musa / Q000737:chemistry*; D010940:Plant Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D010940:Plant Proteins; D007073:Immunoglobulin E; D002688:Chitinases; C119825:chitinase C; D043326:Glucan 1,3-beta-Glucosidase
keywords: Allergens; Banana; Chitinase; Glucanase; Purification; TLP