CHARACTERIZATION OF THE HOUSE-DUST MITE ALLERGEN DER-P-7 BY MONOCLONAL-ANTIBODIES

The house dust mite allergen Der p 7, which was defined by cDNA clonin g, has been shown to react with about 50% of allergic sera and corresp onds to or is antigenically related to at least three different sized components in mite extracts. To characterize these entities, monoclona l antibodies (MoAbs) were generated by immunizing BALB/c mice affinity -purified Der p 7-GST (glutathione S-transferase) fusion protein. MoAb s WH9 and WH22 showed positive reactivity to recombinant Der p 7 negat ive reactivity to GST and the Der p 5-GST fusion protein in ELISA and immunoblotting. The specificity of both MoAbs was confirmed by inhibit ion of the ELISA activity by recombinant Der p 7 but not by the recomb inant Der p 5. Immunoblot analysis demonstrated that both MoAbs showed reactivities to components with molecular weights (mel. wt.) of 31, 3 0 and 26 kDa reactive to both MoAbs. At least six major forms with dif ferent pi or size were indicated by 2-D gel analysis. In addition to c haracterization of Der p 7, both MoAbs may also be considered for use in the standardization of Der p 7 in mite extracts.