Estimation of Der p and Der f I quantities in the reference preparations of Dermatophagoides mite extracts.
A monoclonal antibody-based enzyme-linked immunosorbent assay (MoAb-ELISA) was developed to measure the major Dermatophagoides mite allergens, Der p I and Der f I. The assay was highly species-specific and sensitive. Using this assay system, the absolute mass unit of Der p I and Der f I in the reference preparations of the extracts was estimated. The primary standards used were the purified Der p I and Der f I preparations. The reference preparations of the D. pteronyssinus and D. farinae extracts (92-Dp and 92-Df), which had been prepared from the same amount of mite bodies of both species, were found to contain the same levels of the Der I allergens, 10.1 micrograms/ml of Der p I and 10.0 micrograms/ml of Der f I, respectively. A histamine release assay with leucocytes from mite-allergic donors showed that the total allergenic potency of 92-Dp and 92-Df was comparable. This results indicates that the estimated Der I levels in these extracts seem to be valid, at least, in the balance between the two species, although further comparisons of the absolute quantities by several different laboratories are needed. The Der I levels in the WHO/IUIS international reference preparation of D. pteronyssinus and the CBER standard mite extracts, E4-Dp and E5-Df, were also estimated using this assay system. They were found to contain 4.4 micrograms/vial and 13.3 micrograms/ml of Der p I and 9.5 micrograms/ml of Der f I, respectively.
Yasueda
H
H
Clinical Research Centre for Allergy and Rheumatology, National Sagamihara Hospital, Kanagawa, Japan.
Saito
A
A
Akiyama
K
K
Maeda
Y
Y
Shida
T
T
Sakaguchi
M
M
Inouye
S
S
D000485:Allergens / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000941:Antigens / Q000276:immunology; D039741:Antigens, Dermatophagoides; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006023:Glycoproteins / Q000032:analysis*; D006636:Histamine Release; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D007962:Leukocytes / Q000276:immunology; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D008925:Mites / Q000737:chemistry* / Q000276:immunology; D012015:Reference Standards; D014020:Tissue Extracts; D014944:World Health Organization
D000485:Allergens; D000911:Antibodies, Monoclonal; D000941:Antigens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D014020:Tissue Extracts