BACKGROUND: Group 1 allergens elicit a specific IgE response in about 90% of grass pollen-allergic patients. The aim of this work was to study the antigenic similarity among group 1 allergens from different grasses and to develop a monoclonal antibody (MAb)-based quantitation ELISA. METHODS: Twenty specific MAbs were produced from BALB/c mice immunized with natural Phl p 1. These MAbs were tested for specificity with thirteen different grass pollen extracts from the Poaceae family and in cross-inhibition experiments for the binding of Phl p 1. Purified group 1 allergens from Poeae grasses (Dactylis glomerata, Lolium perenne, Festuca pratensis and Poa pratensis) were tested for parallelism in quantitation ELISA. RESULTS: Eighteen to nineteen anti-Phl p 1 MAbs recognized the homologous allergen in pollen extracts from grasses of the Poeae tribe. In contrast, only four MAbs recognized group 1 from Cynodon dactylon and Phragmites communis. Four groups of MAbs with different epitope specificity were identified. A grass group 1 quantitation ELISA was developed using a mix of three MAbs on the solid phase and a polyclonal rabbit antibody as the second antibody. The group 1 content could be measured in different batches of Phleum pratense as well as in pollen extracts from Poeae grasses, since they showed parallel dose-response curves. CONCLUSIONS: MAbs produced in this work enabled us to show the high antigenic similarity between group 1 allergens from temperate grasses. The results prove the usefulness of the ELISA method developed for standardization of grass allergen products. 2007 S. Karger AG, Basel
BACKGROUND: Group 1 allergens elicit a specific IgE response in about 90% of grass pollen-allergicpatients. The aim of this work was to study the antigenic similarity among group 1 allergens from different grasses and to develop a monoclonal antibody (MAb)-based quantitation ELISA. METHODS: Twenty specific MAbs were produced from BALB/c mice immunized with natural Phl p 1. These MAbs were tested for specificity with thirteen different grass pollen extracts from the Poaceae family and in cross-inhibition experiments for the binding of Phl p 1. Purified group 1 allergens from Poeae grasses (Dactylis glomerata, Lolium perenne, Festuca pratensis and Poa pratensis) were tested for parallelism in quantitation ELISA. RESULTS: Eighteen to nineteen anti-Phl p 1 MAbs recognized the homologous allergen in pollen extracts from grasses of the Poeae tribe. In contrast, only four MAbs recognized group 1 from Cynodon dactylon and Phragmites communis. Four groups of MAbs with different epitope specificity were identified. A grass group 1 quantitation ELISA was developed using a mix of three MAbs on the solid phase and a polyclonal rabbit antibody as the second antibody. The group 1 content could be measured in different batches of Phleum pratense as well as in pollen extracts from Poeae grasses, since they showed parallel dose-response curves. CONCLUSIONS: MAbs produced in this work enabled us to show the high antigenic similarity between group 1 allergens from temperate grasses. The results prove the usefulness of the ELISA method developed for standardization of grass allergen products. 2007 S. Karger AG, Basel
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