Research

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


Reference:

Eriksson, T. L. J., Rasool, O., Huecas, S., Whitley, P., Crameri, R., Appenzeller, U., Gafvelin, G. and 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), pp. 287-294.

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Official URL:

http://dx.doi.org/10.1046/j.1432-1327.2001.01879.x

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.

Details

Item Type Articles
CreatorsEriksson, T. L. J., Rasool, O., Huecas, S., Whitley, P., Crameri, R., Appenzeller, U., Gafvelin, G. and van Hage-Hamsten, M.
DOI10.1046/j.1432-1327.2001.01879.x
DepartmentsFaculty of Science > Biology & Biochemistry
RefereedYes
StatusPublished
ID Code4363

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