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

Eur J Biochem. 2001 Jan;268(2):287-94. doi: 10.1046/j.1432-1327.2001.01879.x.


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 His6-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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / genetics*
  • Allergens / immunology
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Gene Library
  • Humans
  • Immunoglobulin E / blood*
  • Immunoglobulin E / immunology
  • Mites / genetics*
  • Mites / immunology*
  • Molecular Sequence Data
  • Peptide Library
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / immunology
  • Sequence Homology, Amino Acid


  • Allergens
  • Ld 5 allergen
  • Lep d 13 allergen
  • Lep d 7 allergen
  • Peptide Library
  • Recombinant Proteins
  • Immunoglobulin E

Associated data

  • GENBANK/AJ250278
  • GENBANK/AJ250279
  • GENBANK/AJ271058