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Expression of the house dust mite allergen Der p 2 in the baker's yeast Saccharomyces cerevisiae

1998; Wiley; Volume: 28; Issue: 1 Linguagem: Inglês

10.1046/j.1365-2222.1998.00164.x

1365-2222

Hakkaart, Michiel M. Harmsen, Chua, Thomas, Aalberse, van de Ree,

Food Allergy and Anaphylaxis Research

Background and Results The major house dust mite allergen Der p 2 was expressed as a recombinant mature protein in the baker's yeast Saccharomyces cerevisiae . The yeast produces the protein fused to the invertase signal peptide, leading to the secretion of Der p 2 as a soluble protein into the culture medium. The signal peptide is hereby cleaved off, resulting in a mature allergen. In this system Der p 2 was produced in 7.6 (± 2.9) mg/L growth culture. Purification of the recombinant allergen was achieved by a single gel filtration step, resulting in a purity ≥ 95%. The yeast‐derived Der p 2 was almost indistinguishable from natural Der p 2 with respect to IgE‐reactivity and binding to the majority of Der p 2 specific MoAbs — as was shown in RAST analysis ( n = 168) and a sandwich ELISA and RIA analysis, respectively. Recombinant and natural Der p 2 also showed similar biological activity in histamine release assays ( n = 4). Conclusion An expression system for Der p 2 was developed that enables the production of a soluble allergen in the culture supernatant with immunological characteristics similar to the natural allergen. In addition, yeast offers the advantage of the absence of endotoxin in comparison to E. coli . This might facilitate acceptance of recombinant allergens for in vivo applications as immunotherapy or skin‐prick testing.