Monoclonal antibody immunoassay for quantitative analysis of group V allergens in grass pollen extracts |
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Authors: | B. FAHLBUSCH,W.-D. MÜ LLER,G. SCHLENVOIGT,L. JÄ GER,R. WAHL,B. WEBER |
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Affiliation: | Department of Clinical Immunology, The University of Jena, Germany. |
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Abstract: | A two-site monoclonal antibody (MoAb) ELISA has been developed for the quantification of the Phleum pratense major allergen, Phl p V. The assay is based on two MoAbs which recognize different non-overlapping epitopes on the Phl p V molecule; one antibody (1D11) was immobilized on the solid phase and the other (3B2) was biotinylated. An affinity-purified Phl p V preparation (purity of 95%) was used as standard. The assay has a sensitivity of 10 ng/ml of allergen and is suitable for the detection of group V allergen in aqueous grass extracts. The specificity of the assay was investigated with 14 grass pollen and five non-grass pollen extracts. Different levels of group V allergen were detected in extracts of grasses, but not in non-grasses. The assay gives a good correspondence with allergenic activity of extracts as determined by ELISA inhibition using serum pool of allergic patients. The results indicate that the two-site MoAb ELISA could be very useful in the standardization of allergenic extracts from grass pollen. |
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