Determination of Ole e 1 by enzyme immunoassay and scanning densitometry. Validation by skin-prick testing.
Ole e 1 is an important allergen in Olea europaea pollen extracts. This study describes the development of two new methods that can be used to estimate the Ole e 1 content in olive tree pollen extracts. They are based on (1) an enzyme immunoassay that uses rabbit polyclonal, monospecific antibodies and purified Ole e 1, and (2) scanning densitometry of SDS-PAGE gels. Twelve extracts were evaluated by in vivo and in vitro methods. The in vivo biological potency was estimated by prick skin testing 17 allergic individuals; the in vitro allergenic potency by direct IgE and IgE inhibition assays. The enzyme immunoassay showed an operative range of 0.03-100 microg/ml and demonstrated to be specific for Ole e 1. The Ole e 1 content ranged from 1% to 5% of the total protein in the 12 extracts. The amount of Ole e 1, assessed by gel scanning densitometry significantly correlated with the Ole e 1 content obtained by the immunoassay (r = 0.92; p < 0.001). The Ole e 1 content showed a significant correlation with the total allergenic potency of the extracts, evaluated by direct IgE, specific IgE inhibition and skin-prick testing. These two methods can be used to determine the Ole e 1 content in olive pollen extracts. The content of Ole e 1 can vary from 1% to 5% of the total protein in the extracts.
Boluda
L
L
Research and Development Laboratories, C.B.F. LETI, Madrid, Spain.
Sastre
J
J
Casanovas
M
M
Fernández-Caldas
E
E
D000485:Allergens / Q000276:immunology / Q000302:isolation & purification*; D000818:Animals; D000918:Antibody Specificity; D052179:Antigens, Plant; D003720:Densitometry / Q000592:standards; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D007124:Immunoenzyme Techniques / Q000592:standards; D016014:Linear Models; D010940:Plant Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000592:standards; D011817:Rabbits; D015203:Reproducibility of Results; D012882:Skin Tests / Q000592:standards
D000485:Allergens; D052179:Antigens, Plant; D007106:Immune Sera; C081946:Ole e I protein, Olea europaea; D010940:Plant Proteins