Characterization of Dac g 4, a major basic allergen from Dactylis glomerata pollen.
Monoclonal antibodies were produced against Dac g 4, a purified major basic allergen from Dactylis glomerata pollen. Their ability to be used for immunopurification of Dac g 4 was studied on a BIAcore apparatus (Pharmacia). The allergen was purified by affinity chromatography with one monoclonal antibody. Its precise molecular mass, 59,185 +/- 30 d, was determined by mass spectrometry. Its isoelectric point is 10.4. Sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoblotting showed that Dac g 4-related proteins of similar molecular mass were detected in the majority of allergenic grass pollen species. By double-site ELISAs, we have estimated that Dac g 4 represents about 6% of the total proteins from a water-soluble extract. One monoclonal antibody (mAb H) recognized a 60 kd cross-reactive protein in other grass pollens, though none in any of the tree or weed pollens tested. Inhibition studies of IgE antibody binding to Dac g 4 with pollen extracts confirmed the presence of cross-reactive allergens in Secale cereale, Lolium perenne, Festuca elatior, Holcus lanatus, Bromus arvensis, Poa pratense, Hordeum sativum, and Phleum pratense.
Leduc-Brodard
V
V
Unité d'Immuno-Allergie, Institut Pasteur, Paris, France.
Inacio
F
F
Jaquinod
M
M
Forest
E
E
David
B
B
Peltre
G
G
D000485:Allergens / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D015374:Biosensing Techniques; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification*; D011058:Pollen / Q000737:chemistry* / Q000276:immunology
D000485:Allergens; D000596:Amino Acids; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; C103001:Dac g 4 protein, Dactylis glomerata; D010940:Plant Proteins