Identification of IgE-binding egg white proteins: comparison of results obtained by different methods.
The binding of IgE to egg white proteins was investigated for 34 sera from adults with a positive case history and/or positive RAST towards egg, and the impact of experimental conditions on IgE binding in commonly used methods was studied. Radioimmunoblotting after SDS-PAGE of both reduced and unreduced egg white extracts showed complex reaction patterns. The results were confirmed by crossed radioimmunoelectrophoresis (CRIE). Radio dot immunobinding was used to investigate the effect of treatment of allergens for SDS-PAGE and to evaluate the other methods. As a conclusion, the use of combinations of at least two methods is recommended for the identification of IgE-binding egg white proteins. Of the 34 sera, 18 reacted with ovotransferrin, 13 with ovomucoid, 11 with ovalbumin and 5 with lysozyme. The amounts of IgE bound to ovalbumin and lysozyme were generally lower than the amounts bound to ovotransferrin and ovomucoid.
Aabin
B
B
Department of Biochemistry and Nutrition, Technical University of Denmark, Lyngby.
Poulsen
L K
LK
Ebbehøj
K
K
Nørgaard
A
A
Frøkiaer
H
H
Bindslev-Jensen
C
C
Barkholt
V
V
D000485:Allergens / Q000032:analysis*; D000818:Animals; D000941:Antigens / Q000032:analysis; D001345:Autoradiography; D002645:Chickens; D003378:Counterimmunoelectrophoresis / Q000379:methods; D004527:Egg Proteins / Q000276:immunology / Q000378:metabolism*; D004531:Eggs / Q000009:adverse effects*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005512:Food Hypersensitivity / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007073:Immunoglobulin E / Q000276:immunology / Q000378:metabolism*; D011817:Rabbits; D011852:Radioallergosorbent Test / Q000379:methods
D000485:Allergens; D000941:Antigens; D004527:Egg Proteins; D007073:Immunoglobulin E