Cloning of three new allergens from the dust mite Lepidoglyphus destructor using phage surface display technology

T L J Eriksson, O Rasool, S Huecas, P Whitley, R Crameri, U Appenzeller, G Gafvelin, M van Hage-Hamsten

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42 Citations (Scopus)

Abstract

The dust mite Lepidoglyphus destructor is a common species in Europe and a major cause of dust mite allergy in rural surroundings, but it also contributes to dust mite allergy in urban areas. One major allergen, Lep d 2, has been expressed as a recombinant protein and evaluated both in vivo and in vitro and shown to detect 60% or more of L. destructor-sensitized subjects. Additional recombinant allergens are needed to obtain a reliable diagnostic tool for L. destructor allergy. The aim of this study was to clone and express new allergens from L. destructor and determine their recognition frequency among sensitized individuals. A phage display cDNA expression library was constructed and screened with sera from L. destructor-sensitized individuals. The cDNAs encoding the allergens were cloned into the pET17b vector and subsequently expressed in Escherichia coli as C-terminal His(6)-tagged proteins. Immunoblotting of the recombinant proteins was performed using sera from 45 subjects allergic to L. destructor. Three new allergens from L. destructor, Ld 5 (originating from a partial Lep d 5 clone), Lep d 7 and Lep d 13, were identified and recognized by 4/45 (9%), 28/45 (62%) and 6/45 (13%) sera from L. destructor-sensitized subjects, respectively.
Original languageEnglish
Pages (from-to)287-294
Number of pages8
JournalEuropean Journal of Biochemistry
Volume268
Issue number2
DOIs
Publication statusPublished - Jan 2001

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    Eriksson, T. L. J., Rasool, O., Huecas, S., Whitley, P., Crameri, R., Appenzeller, U., Gafvelin, G., & van Hage-Hamsten, M. (2001). Cloning of three new allergens from the dust mite Lepidoglyphus destructor using phage surface display technology. European Journal of Biochemistry, 268(2), 287-294. https://doi.org/10.1046/j.1432-1327.2001.01879.x