Mapping of Dermatophagoides farinae mite allergens by two-dimensional immunoblotting

J Allergy Clin Immunol. 1998 Oct;102(4 Pt 1):631-6. doi: 10.1016/s0091-6749(98)70280-5.


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.

Conclusion: 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.

MeSH terms

  • Allergens / chemistry*
  • Amino Acid Sequence
  • Animals
  • Antigens, Dermatophagoides
  • Chitinases / chemistry
  • Electrophoresis, Gel, Two-Dimensional
  • Glycoproteins / chemistry
  • Humans
  • Immunosorbent Techniques
  • Isoelectric Focusing
  • Mites / immunology*
  • Molecular Sequence Data
  • Molecular Weight
  • Peptide Mapping


  • Allergens
  • Antigens, Dermatophagoides
  • Glycoproteins
  • Chitinases