Mapping of fruit allergens by 2D electrophoresis and immunodetection.
Proteomic analyses of fruits are confronted with a series of specific obstacles: a general low protein content in plant tissues, allergen extraction from highly complex matrices and protein determination in the presence of interfering compounds. Different methods are currently being introduced to achieve higher protein yields and a simultaneous removal of interfering substances, such as polyphenols and polysaccharides. However, no universal protocol suitable for protein purification from any given plant species is available. Protein profiling by 2DE-western blotting offers a powerful tool for the detection and characterization of known and novel plant allergens. Moreover, the detection of IgE-reactive proteins from fruits is improved by combining western blot and alternative visualization techniques. The recent developments in bioinformatics and databases facilitate the interpretation of profiling studies with regard to novel potential fruit allergens.
Marzban
Gorji
G
Institute of Applied Microbiology, Plant Biotechnology Unit, Department of Biotechnology, BOKU University, Muthgasse 18, House B Vienna, A-1190, Austria. gorji.marzban@boku.ac.at
Herndl
Anita
A
Maghuly
Fatemeh
F
Katinger
Hermann
H
Laimer
Margit
M
D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D052179:Antigens, Plant / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western / Q000379:methods*; D003429:Cross Reactions; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D000939:Epitopes / Q000737:chemistry; D005512:Food Hypersensitivity / Q000175:diagnosis / Q000209:etiology; D005638:Fruit / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis / Q000276:immunology; D007457:Iodine Radioisotopes / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D011058:Pollen / Q000276:immunology; D012698:Serologic Tests / Q000379:methods
D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D000939:Epitopes; D007457:Iodine Radioisotopes; D010940:Plant Proteins; D007073:Immunoglobulin E