Monoclonal antibody immunoassay for quantitative analysis of group V allergens in grass pollen extracts.
A two-site monoclonal antibody (MoAb) ELISA has been developed for the quantification of the Phleum pratense major allergen, Phl p V. The assay is based on two MoAbs which recognize different non-overlapping epitopes on the Phl p V molecule; one antibody (1D11) was immobilized on the solid phase and the other (3B2) was biotinylated. An affinity-purified Phl p V preparation (purity of 95%) was used as standard. The assay has a sensitivity of 10 ng/ml of allergen and is suitable for the detection of group V allergen in aqueous grass extracts. The specificity of the assay was investigated with 14 grass pollen and five non-grass pollen extracts. Different levels of group V allergen were detected in extracts of grasses, but not in non-grasses. The assay gives a good correspondence with allergenic activity of extracts as determined by ELISA inhibition using serum pool of allergic patients. The results indicate that the two-site MoAb ELISA could be very useful in the standardization of allergenic extracts from grass pollen.
Fahlbusch
B
B
Department of Clinical Immunology, The University of Jena, Germany.
Müller
W D
WD
Schlenvoigt
G
G
Jäger
L
L
Wahl
R
R
Weber
B
B
D000485:Allergens / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology*; D019921:Biotinylation; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D010936:Plant Extracts / Q000032:analysis / Q000276:immunology*; D010940:Plant Proteins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification / Q000592:standards; D006109:Poaceae; D011058:Pollen / Q000276:immunology*; D012015:Reference Standards; D015203:Reproducibility of Results; D006255:Rhinitis, Allergic, Seasonal / Q000097:blood / Q000276:immunology; D012680:Sensitivity and Specificity
D000485:Allergens; D000911:Antibodies, Monoclonal; D000939:Epitopes; C076049:Phl p V protein, Phleum pratense; D010936:Plant Extracts; D010940:Plant Proteins