Abstract
Rationale Tropomyosins derived from invertebrates are a well-known pan allergen. However, the allergenicity of the recombinant tropomyosins is not coherent. The aim of this study was to compare the IgE-binding reactivities of native and recombinant German cockroach tropomyosins. Methods Native tropomyosin was purified by ammonium sulfate fractionation, hydroxyapatite column chromatography, and gel elution. Recombinant tropomyosin was expressed in Pichia pastoris. The allergenicity of native and recombinant tropomyosin was compared by ELISA inhibition study. Results Native German cockroach tropomyosin showed 17.6% IgE-binding reactivity from German cockroach sensitized sera. Recombinant tropomyosin was produced without fusion protein and the N-terminus was blocked as a native counterpart. Nevertheless, the IgE binding reactivity was comparable to that of native tropomyosin over the concentration range of 1 to 1000 ng/ml concentration in the ELISA inhibition test. Conclusions Recombinant German cockroach tropomyosin that was improved in terms of allergenicity relative to the recombinant protein expressed in Escherichia coli. Other factors, other than the structural differences of native and recombinant proteins, could influence on the various IgE reactivities of tropomyosins.
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