BACKGROUND: The major house dust mite Der p 1 allergen is associated with @entity56 . Heterologous over-expression of biologically active Der p 1 was previously attempted but with limited success. OBJECTIVE: The aim of this study was to establish an efficient system for the production of recombinant Der p 1. METHODS: The proform of Der p 1 was expressed in @entity10056 as a fusion with the alpha mating factor signal sequence. The recombinant product was purified from culture medium and compared to Der p 1 isolated from mite culture, in terms of enzymatic activity as well as IgE binding capacity. RESULTS: ProDer p 1 was efficiently secreted into culture medium as a hyperglycosylated protein of 40-60 kDa. Postpurification dialysis in acidic buffer was required for the autocatalytic processing of Der p 1. During this treatment, the prosequence was efficiently removed to give highly glycosylated recombinant mature Der p 1. Competition ELISA experiments as well as @entity462 proteinase activity assays indicated that recombinant processed Der p 1 was similar to natural Der p 1 isolated from mite cultures in terms of IgE binding and enzymatic activities. However, the @entity112 releasing activity of recombinant Der p 1 was slightly weaker than that of natural Der p 1. CONCLUSION: This efficient system for recombinant Der p 1 expression leads the way for the design of new diagnostics for house dust mite @entity566 , epitope mapping, allergen engineering, structural and immunological studies and new immunotherapeutic treatments.

High-level expression of recombinant house dust mite allergen Der p 1 in XXXX .

@entity462 :: ('MESH:D003545', 'Chemical') :: ['cysteine'] @entity10056 :: ('4922', 'Species') :: ['Pichia pastoris'] @entity566 :: ('MESH:D006967', 'Disease') :: ['allergy'] @entity56 :: ('MESH:D004342', 'Disease') :: ['allergic disease'] @entity112 :: ('MESH:D006632', 'Chemical') :: ['histamine']
@entity10056 :: (4922,Species) :: ['Pichia pastoris']
