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Mapping of Malus domestica allergens by 2‐D electrophoresis and IgE‐reactivity
Author(s) -
Herndl Anita,
Marzban Gorji,
Kolarich Daniel,
Hahn Rainer,
Boscia Donato,
Hemmer Wolfgang,
Maghuly Fatemeh,
Stoyanova Elitza,
Katinger Hermann,
Laimer Margit
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600342
Subject(s) - allergen , polyclonal antibodies , immunoglobulin e , blot , epitope , chemistry , antibody , epitope mapping , immunology , allergy , biology , microbiology and biotechnology , biochemistry , gene
Abstract The importance of apple allergens has been repeatedly emphasized, and their presence has been confirmed both in pollen and in fruits. In the present study, a combination of proteomic tools have been used to build a complete allergen map of apple. The water‐soluble fraction of an apple extract was precipitated using a phenol‐based procedure and separated by 2‐DE. Initially four previously classified allergens, Mal d 1, Mal d 2, Mal d 3 and Mal d 4, could be identified in Western blots with polyclonal rabbit antibodies directed to the four respective allergens, and subsequently matched to the bands recognized by several patient sera. Further, all four known apple allergens were localized on a 2‐DE map and they were matched with spots recognized by sera of patients with different allergic patterns. Moreover, a new, putative allergen could be identified using MS. We evaluated the influence of post‐translational modifications and the immunoreactivity under different analytical conditions. The comparison of different visualization methods for 2‐DE gels and blots revealed that even very low concentrations of the intact epitopes are detectable by IgEs of patients, and therefore might be sufficient to trigger allergic symptoms in sensitized individuals.