Luminescence immunoassay of pollen allergens on air sampling polytetrafluoroethylene filters

Abstract

A new method for quantification of airborne birch and grass pollen allergens collected on porous polytetrafluoroethylene filters has been developed. In this method, the allergens firmly adsorbed to the sampling filter of 25 mm in diameter are reacted with specific antibodies conjugated to alkaline phosphatase, generating a matrix-bound allergen–antibody–phosphatase complex. The filter is then floated on a chemiluminescent enzyme substrate solution. The light intensity of the product is linearly related to the amount of allergen over a large mass range, 0–1000 SQ (1 SQ is about 250 pg of protein). This direct on sampling filter in solution (DOSIS) technique demonstrated intra-assay precisions between 6–16% and 11–15% for the levels of 1–100 SQ units of grass allergen Phl p 5 and 4–400 SQ units of birch allergen Bet v 1, respectively. The limits of quantification for the corresponding allergens were estimated to 0.5 and 2 SQ units. Application of DOSIS to analysis of the grass pollen allergen concentrations of outdoor air for 12 days in July 1998 revealed a correlation coefficient of 0.69 between pollen grain and allergen concentrations for the dry weather period. After rainy days large amounts of grass allergens were present even in the absence of pollen grains.