A two-site monoclonal antibody ELISA for the quantification of group V allergens in grass extracts

A two-site solid-phase enzymimmunoassay was used for the quantification of group V allergen in grass pollen extracts in mass units. The assay is based on the monoclonal antibodies (MoAbs), 1D11 and 3B2 which recognize different epitopes on the standard Phl p V. The MoAb-ELISA is very sensitive (15-100 ng/ml Phl p V) and highly specific for group V allergens. Six pollen extracts of different grasses demonstrated parallel binding curves. The group V content ranged between 73 and 673 micrograms/ml in the extracts. The International Standard of Phleum pratense (IS 82/520) contains 400 micrograms/ml Phl p V. A good correlation was observed between the group V content and RAST inhibition, with the exception of Poa pratensis.