title: A proteomic style approach to characterize a grass mix product reveals potential immunotherapeutic benefit.
abstract: BACKGROUND
  Grass allergy immunotherapies often consist of a mix of different grass extracts, each containing several proteins of different physiochemical properties; however, the subtle contributions of each protein are difficult to elucidate. This study aimed to identify and characterize the group 1 and 5 allergens in a 13 grass extract and to standardize the extraction method.
  
  
  METHODS
  The grass pollens were extracted in isolation and pooled and also in combination and analyzed using a variety of techniques including enzyme-linked immunosorbent assay, liquid chromatog-raphy-mass spectrometry, and sodium dodecyl sulfate-polyacrylam-ide gel electrophoresis.
  
  
  RESULTS
  Gold-staining and IgE immunoblotting revealed a high degree of homology of protein bands between the 13 species and the presence of a densely stained doublet at 25-35 kD along with protein bands at approximately 12.5, 17, and 50 kD. The doublet from each grass species demonstrated a high level of group 1 and 5 interspecies homology. However, there were a number of bands unique to specific grasses consistent with evolutionary change and indicative that a grass mix immunotherapeutic could be considered broad spectrum.
  
  
  CONCLUSIONS
  Sodium dodecyl sulfate-polyacrylamide gel electro-phoresis and IgE immunoblotting showed all 13 grasses share a high degree of homology, particularly in terms of group 1 and 5 allergens. IgE and IgG enzyme-linked immunosorbent assay potencies were shown to be independent of extraction method.
authors.affiliation: Allergy Therapeutics, Dominion Way, Worthing, West Sussex, 8SA, UK.
authors.forename: Alan
authors.initials: A
authors.lastname: Bullimore
authors.forename: Nicola
authors.initials: N
authors.lastname: Swan
authors.forename: Wemimo
authors.initials: W
authors.lastname: Alawode
authors.forename: Murray
authors.initials: M
authors.lastname: Skinner