title: Characterization of recombinant cat albumin.
abstract: BACKGROUND
  Indoor allergens derived from animals and mites often contribute to exacerbation of skin manifestations in atopic dermatitis (AD) patients.
  
  
  OBJECTIVE
  To produce and characterize recombinant cat albumin, a cross-reactive animal allergen.
  
  
  METHODS
  A complete cDNA coding for cat albumin was obtained by RT-PCR amplification from cat liver RNA. Recombinant cat albumin was expressed in Escherichia coli as hexahistidine-tagged protein, purified by nickel affinity chromatography and studied for IgE reactivity with sera from cat-allergic patients by ELISA and immunoblotting. Furthermore, CD203c expression of basophils from cat-allergic patients upon exposure to recombinant cat albumin was analysed.
  
  
  RESULTS
  Recombinant cat albumin, a cross-reactive animal allergen sharing most IgE epitopes with its natural counterpart, was produced in E. coli. It was recognized preferentially by IgE from AD patients and elicited IgE-dependent basophil activation in sensitized patients.
  
  
  CONCLUSIONS
  Recombinant cat albumin may be used as a paradigmatic tool to analyse mechanisms of allergen-triggered exacerbation of AD, for diagnostic and, perhaps for therapeutic purposes.
authors.affiliation: Institute of Medical and Chemical Laboratory Diagnostics, University of Vienna, Vienna, Austria.
authors.forename: R
authors.initials: R
authors.lastname: Reininger
authors.forename: I
authors.initials: I
authors.lastname: Swoboda
authors.forename: B
authors.initials: B
authors.lastname: Bohle
authors.forename: A W
authors.initials: AW
authors.lastname: Hauswirth
authors.forename: P
authors.initials: P
authors.lastname: Valent
authors.forename: H
authors.initials: H
authors.lastname: Rumpold
authors.forename: R
authors.initials: R
authors.lastname: Valenta
authors.forename: S
authors.initials: S
authors.lastname: Spitzauer
mesh_terms: D000418:Albumins / Q000235:genetics* / Q000276:immunology; D000485:Allergens / Q000235:genetics* / Q000276:immunology; D000818:Animals; D015549:Basophil Degranulation Test; D002415:Cats / Q000276:immunology*; D003429:Cross Reactions; D018076:DNA, Complementary / Q000032:analysis; D003876:Dermatitis, Atopic / Q000175:diagnosis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases / Q000032:analysis; D011755:Pyrophosphatases / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000418:Albumins; D000485:Allergens; D018076:DNA, Complementary; C427944:ENPP3 protein, human; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases