Quantification of group 1 and group 5 allergens in extracts of several grass species

Quantification and detection of major allergens in complex allergen extract s by specific monoclonal antibodies (mabs) has proved to be highly sensitiv e and precise. Group 1 and group 5 allergens have been defined and characte rized as major allergens of grass pollen extracts, and the quantification o f group 5 allergens has been described previously [5]. Lol p I specific mab s have now been produced and characterized Two of these mabs cross-react wi th group I allergens of other grass species (Dactylis glomerata, Festuca pr atensis, Holcus lanatus, Poa pratensis and Phleum pratense). Both mabs bind to non-allergens. Detailed knowledge of the composition of grass pollen ex tracts is essential to ensure the production of high quality extracts, and this can be improved by the quantification of group 1 and 5 allergens. Furt hermore, the data will help in a better understanding of the relationship b etween major allergen content and biological activity of allergen extracts. overlapping epitopes. A two-site-binding ELISA was established using mab 3G 5 for capture and mab 9C12 for detection of Lol p 1 and the resulting stand ard curve gave a range of detection between 25 ng and 500 ng Lol p 1/ml. Bi nding curves es of Lol p 1 and group 1 allergens of other grass species wer e parallel. Group 1 and 5 allergens were quantified in extracts of each of the 6 grasses and in an extract of a grass pollen mixture. Group 1 and 5 co ntents vary indicating species specific differences. Group I contents were de termined between 148 mu g/ml (Phleum pratense) and 630 mu g/ml (Lolium p erenne), and group 5 contents between 8.3 mu g/ml (Poa pratense) and 76 mu g/ml (Lolium perenne). Furthermore, species-specific differences were detec ted in the ratios of these major Quantification and detection of major alle rgens in complex allergen extracts by specific monoclonal antibodies (mabs) has proved to be highly sensitive and precise. Group 1 and group 5 allerge ns have been defined and characterized as major allergens of grass pollen e xtracts, and the quantification of group 5 allergens has been described pre viously [5]. Lol p I specific mabs have now been produced and characterized Two of these mabs cross-react with group I allergens of other grass specie s (Dactylis glomerata, Festuca pratensis, Holcus lanatus, Poa pratensis and Phleum pratense). Both mabs bind to nonoverlapping epitopes. A two-site-bi nding ELISA was established using mab 3G5 for capture and mab 9C12 for dete ction of Lol p 1 and the resulting standard curve gave a range of detection between 25 ng and 500 ng Lol p 1/ml. Binding curves es of Lol p 1 and grou p 1 allergens of other grass species were parallel. Group 1 and 5 allergens were quantified in extracts of each of the 6 grasses and in an extract of a grass pollen mixture. Group 1 and 5 contents vary indicating species spec ific differences. Group I contents were de termined between 148 mu g/ml (Ph leum pratense) and 630 mu g/ml (Lolium perenne), and group 5 contents betwe en 8.3 mu g/ml (Poa pratense) and 76 mu gml (Lolium perenne). Furthermore, species-specific differences were detected in the ratios of these major all ergens. Detailed knowledge of the composition of grass pollen extracts is e ssential to ensure the production of high quality extracts, and this can be improved by the quantification of group 1 and 5 allergens. Furthermore, th e data will help in a better understanding of the relationship between majo r allergen content and biological activity of allergen extracts.