title: Mapping of Dermatophagoides farinae mite allergens by two-dimensional immunoblotting.
abstract: BACKGROUND
  Allergens from the house dust mite Dermatophagoides farinae are responsible for frequent respiratory allergic disorders, but only 3 groups of these allergens are well characterized.
  
  
  OBJECTIVE
  This study was performed to complete the repertoire of D farinae allergens using two-dimensional (2-D) electrophoresis.
  
  
  METHODS
  D farinae mite allergens, extracted from whole cultures in the presence of a mild detergent, were separated by 2-D electrophoresis with subsequent immunoblotting. IgE-binding proteins were detected with individual mite-sensitive patient sera and the anti-D pteronyssinus human serum pool. Allergens were identified by an inhibition immunoblot test, by means of specific mAbs, or by biochemical characterization. The internal peptides of 2 allergens were microsequenced.
  
  
  RESULTS
  2-D immunoblotting with individual patient sera showed a marked heterogeneity in the isoelectric point of the allergens, as well as differences in the individual IgE-binding patterns. In addition to identification of allergens Der f 1, Der f 2, and Der f 3, new allergens have been characterized as Der f 4, Der f 5, and 2 high molecular mass allergens. Microsequencing of peptides from the latter allergens revealed significant homologies with allergen Mag 3 from D farinae and with a chitinase from prawn Penaeus japonicus.
  
  
  CONCLUSIONS
  2-D electrophoresis with subsequent immunoblotting and protein microsequencing allowed characterization of a more complete repertoire of D farinae allergens and their multiple isoforms, and identification of six new allergens.
authors.affiliation: Unité d'Immuno-Allergie, Institut Pasteur, Paris, France.
authors.forename: J
authors.initials: J
authors.lastname: Le Mao
authors.forename: C E
authors.initials: CE
authors.lastname: Mayer
authors.forename: G
authors.initials: G
authors.lastname: Peltre
authors.forename: F X
authors.initials: FX
authors.lastname: Desvaux
authors.forename: B
authors.initials: B
authors.lastname: David
authors.forename: A
authors.initials: A
authors.lastname: Weyer
authors.forename: H
authors.initials: H
authors.lastname: Sénéchal
mesh_terms: D000485:Allergens / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D039741:Antigens, Dermatophagoides; D002688:Chitinases / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D007163:Immunosorbent Techniques; D007525:Isoelectric Focusing; D008925:Mites / Q000276:immunology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins; D002688:Chitinases