Expression and biologic activity of recombinant allergens

Traditional methods of allergy diagnosis and treatment have relied on the use of heterogeneous allergen extracts. The problems with these extracts are that the absolute potency is difficult to define and some patients give adverse reactions when the extracts are injected (during immunotherapy), which can be life threatening. While there has been significant progress in the standardization of allergen extracts; the use of purified recombinant allergens expressed in bacteria, yeast or insect cells offers the prospect of using defined proteins for both diagnosis and treatment. An increasing number of recombinant allergens have been produced and it is possible to investigate whether cocktails of 3-4 allergens would be suitable for clinical purposes. Typical vectors in which allergens have been expressed include pGEX and pET (in Escherichia coii); pSAY-1 and Pichia pastoris (yeast), and the baculovirus system, giving yields of 1-10 mg 1 1. The allergens can be obtained with a high degree of purity and, in many cases, have excellent immunoreactivity on skin testing and in serum IgE antibody assays. Several recombinant allergens from mite (Der p 2, Bio t 5, Der p 5), aspergillus (Asp f 1), cockroach (Bla g 4, Bla g 5) and grasses (Phi p 1, Phi p 5) have been shown to have good biologic activity on skin testing, and elicit positive wheal and flare reactions using pico-gram doses of protein.

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