Species-specific measurement of the second group of Dermatophagoides mite allergens, Der p 2 and Der f 2, using a monoclonal antibody-based ELISA.
BACKGROUND
The group 2 Dermatophagoides mite allergens, Der p 2 and Der f 2, were known to be highly crossreactive, and previous assays to measure Der p 2 and Der f 2 were not species-specific.


OBJECTIVE
The aim of this study was to develop a monoclonal antibody-based ELISA (MoAb-ELISA) to species-specifically measure Der p 2 and Der f 2.


METHODS
The MoAb-ELISA for Der p 2 and Der f 2 was performed using species-specific MoAbs for Der p 2 and Der f 2 and a biotinylated second MoAb which recognized a common epitope on both Der p 2 and Der f 2.


RESULTS
The assay was highly species-specific, reproducible and sensitive. Thirty-two house dust samples were assayed by the MoAb-ELISA for Der p 2 and Der f 2 and by a previously reported radioimmunoassay for Der 2 with rabbit anti-Der 2 antibodies. The summed values for Der p 2 and Der f 2 by the MoAb-ELISA demonstrated a good correlation with the Der 2 values using the radioimmunoassay (r = 0.978). Furthermore, the proportion of the Der p 2 level in the total Der 2 level (Der p 2 divided by Der p 2 plus Der f 2) correlated well with that of the D. pteronyssinus mite number to the total Dermatophagoides mite number identified by species (r = 0.970).


CONCLUSIONS
The MoAb-ELISA for Der p 2 and Der f 2, as well as that of Der p 1 and Der f 1, will be useful for the standardization of mite extracts and for the assessments of mite allergen exposure.
Yasueda
H
H
Clinical Research Center for Allergy and Rheumatology, National Sagamihara Hospital, Sagamihara, Japan.
Saito
A
A
Yanagihara
Y
Y
Akiyama
K
K
Takaoka
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000941:Antigens / Q000032:analysis*; D039741:Antigens, Dermatophagoides; D004391:Dust; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000032:analysis*; D006825:Hybridomas; D051379:Mice; D008925:Mites; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D000941:Antigens; D039741:Antigens, Dermatophagoides; D004391:Dust; D006023:Glycoproteins