title: Immune-reactivity of recombinant isoforms of the major house dust mite allergen Der p 2.
abstract: BACKGROUND
  Recombinant Der p 2, expressed in yeast, lacked reactivity with 5 monoclonal antibodies against natural Der p 2.
  
  
  OBJECTIVE
  The aim of this study was to investigate whether the lack of reactivity with recombinant Der p 2 can be explained by the existence of isoforms.
  
  
  METHODS
  By site-directed mutagenesis three recombinant isoforms of Der p 2 were produced. Reactivity with monoclonal antibodies and human IgE was analysed by means of RAST and RAST-inhibition.
  
  
  RESULTS
  All five monoclonals that lacked reactivity with the originally selected isoform, showed reactivity upon replacement of aspartic acid by asparagine at position 114. The other two substitutions (at position 26 and 47) had no effect. Binding of human IgE (n = 10) was not significantly influenced by the isogenetic variation at position 114.
  
  
  CONCLUSIONS
  Monoclonal antibodies raised against natural Der p 2 can sometimes discriminate between different isoforms, allowing the study of the natural occurrence of isoforms. For application in allergen-measurement assays, non-discriminating monoclonal antibodies should be selected.
authors.affiliation: Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, University of Amsterdam.
authors.forename: G A
authors.initials: GA
authors.lastname: Hakkaart
authors.forename: M D
authors.initials: MD
authors.lastname: Chapman
authors.forename: R C
authors.initials: RC
authors.lastname: Aalberse
authors.forename: R
authors.initials: R
authors.lastname: van Ree
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D039741:Antigens, Dermatophagoides; D004391:Dust; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D014644:Genetic Variation; D006023:Glycoproteins / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D008925:Mites / Q000276:immunology*; D016297:Mutagenesis, Site-Directed / Q000276:immunology; D011852:Radioallergosorbent Test; D011994:Recombinant Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D039741:Antigens, Dermatophagoides; D004391:Dust; D000939:Epitopes; D006023:Glycoproteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E