Novel monoclonal antibodies against house dust mite allergen Der p 21
and their application to analyze allergen extracts
Allergen extracts and recombinant allergens are used in allergy
diagnostics and immunotherapy. Since allergen extracts from different
manufacturers lack proper standardization regarding their composition,
monoclonal antibodies (MAbs) against specific allergen components can be
used for their identification and quantification in allergen extracts.
This study aimed to generate MAbs against allergen Der p 21 of
Dermatophagoides pteronyssinus for the analysis of allergen extracts.
Recombinant Der p 21 was expressed in E. coli and purified using
affinity chromatography. MAbs against Der p 21 were generated using
hybridoma technology. House dust mite (HDM) allergen extracts were
analyzed using the newly developed sandwich enzyme-linked immunosorbent
assay, Western blotting and microarray immunoassay. MAbs were proven to
be reactive with natural Der p 21. Highly specific sandwich
enzyme-linked immunosorbent assay for the quantification of Der p 21 was
developed. The allergen was detected and its concentration was
determined in only 3 of 6 analyzed HDM allergen extracts from different
manufacturers. HDM analysis by MAb-based immunoassays shows their
differences in allergen composition. The results demonstrate the
importance of allergen-specific MAbs as a tool for the characterization
of allergen extracts and the need for their appropriate standardization
before their use for allergy diagnostics or immunotherapy.