Human monoclonal antibody–based quantification of group 2 grass pollen allergens Katharina Marth, MD, Margarete Focke, PhD, Sabine Flicker, PhD, Rudolf.

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Human monoclonal antibody–based quantification of group 2 grass pollen allergens Katharina Marth, MD, Margarete Focke, PhD, Sabine Flicker, PhD, Rudolf Valenta, MD Journal of Allergy and Clinical Immunology Volume 113, Issue 3, Pages 470-474 (March 2004) DOI: 10.1016/j.jaci.2003.11.042

FIG 1 Cross-reactivity of human monoclonal anti–Phl p 2 antibody with group 2 pollen allergens from different grass species. Nitrocellulose membranes containing pollen extracts from 9 grasses with (2: Phleum pratense; 3: Poa pratensis; 4: Dactylis glomerata; 5: Triticum aestivum; 6: Avena sativa; 7: Anthoxanthum odoratum; 8: Hordeum vulgare; 9: Lolium perenne), one (10: Cynodon dactylon) without group 2 allergens, rPhl p 2 (lane 1) and birch pollen extract (lane 11: negative control) were incubated with the anti–Phl p 2 antibody. Molecular weights are indicated (in kd) on the right margin; position of group 2 allergens is indicated. Journal of Allergy and Clinical Immunology 2004 113, 470-474DOI: (10.1016/j.jaci.2003.11.042)

FIG 2 Description of the ELISA for quantification of group 2 grass pollen allergens. A, ELISA plates are coated with the huMab2 and B, incubated with a mixture containing defined amounts of biotinylated rPhl p 2 (competitor: blue dots with yellow asterisks) and unknown contents of group 2 allergens (blue dots). C, Labeled rPhl p 2 is detected with horseradish peroxidase–conjugated streptavidin (red) and D, visualized by color reaction (green) by using 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). The intensity of the color reaction is inversely related to the concentration of group 2 allergens. According to a standard curve established with defined amounts of unlabeled rPhl p 2, the concentration of group 2 allergens can be determined. Journal of Allergy and Clinical Immunology 2004 113, 470-474DOI: (10.1016/j.jaci.2003.11.042)

FIG 3 Dose-response curves of rPhl p 2 determined by competition ELISA. A, Interassay standard deviations (means of 6 determinations) and B, intra-assay standard deviations (means of 5 determinations) shown for different concentrations of rPhl p 2 (x-axis: logarithmic values). Journal of Allergy and Clinical Immunology 2004 113, 470-474DOI: (10.1016/j.jaci.2003.11.042)