2-DIMENSIONAL GEL-ELECTROPHORETIC ANALYSIS WITH IMMOBILIZED PH GRADIENTS OF DACTYLIS-GLOMERATA POLLEN ALLERGENS

We studied Orchard grass pollen allergens (Dactylis glomerata) by a ne w two-dimensional electrophoretic technique which is the combination o f isoelectric focusing (IEF) in immobilized pH gradient (IPG) and hori zontal SDS-PAGE. The IPG-Dalt (two-dimensional gel electrophoresis wit h immobilized pH gradients in the first dimension) electrophoresis was made with four different but complementary pH gradients: 3-10.5, 4-9, 4-7, 6.3-10.5. We also compared the behavior of allergens in native a nd denaturing conditions during IEF. This technique followed bye elect roblotting on nitrocellulose membrane and immunodetection with an alle rgic patient serum was shown to increase the number of IgE-binding com ponents up to 100 compared to one-dimensional techniques. Major allerg en groups could be identified. New allergens not yet described could b e characterized by their isoelectric points and their molecular masses . This highly resolving and reproducible technique, should allow an ea sy allergen standardization.