title: The influence of recombinant production on the immunologic behavior of birch pollen isoallergens.
abstract: BACKGROUND
  Allergic reactions towards the birch major pollen allergen Bet v 1 are among the most common causes of spring pollinosis in the temperate climate zone of the Northern hemisphere. Natural Bet v 1 is composed of a complex mixture of different isoforms. Detailed analysis of recombinant Bet v 1 isoforms revealed striking differences in immunologic as well as allergenic properties of the molecules, leading to a classification of Bet v 1 isoforms into high, medium, and low IgE binding proteins. Especially low IgE binding Bet v 1 isoforms have been described as ideal candidates for desensitizing allergic patients with allergen specific immunotherapy (SIT). Since diagnosis and therapy of allergic diseases are highly dependent on recombinant proteins, continuous improvement of protein production is an absolute necessity.
  
  
  METHODS
  Therefore, two different methods for recombinant production of a low IgE binding Bet v 1 isoform were applied; one based on published protocols, the other by implementing latest innovations in protein production. Both batches of Bet v 1.0401 were extensively characterized by an array of physicochemical as well as immunological methods to compare protein primary structure, purity, quantity, folding, aggregation state, thermal stability, and antibody binding capacity.
  
  
  CONCLUSIONS
  The experiments demonstrated that IgE antibody binding properties of recombinant isoallergens can be significantly influenced by the production method directly affecting possible clinical applications of the molecules.
authors.affiliation: Christian Doppler Laboratory for Allergy Diagnosis and Therapy, University of Salzburg, Salzburg, Austria. michael.wallner@sbg.ac.at
authors.forename: Michael
authors.initials: M
authors.lastname: Wallner
authors.forename: Martin
authors.initials: M
authors.lastname: Himly
authors.forename: Angela
authors.initials: A
authors.lastname: Neubauer
authors.forename: Anja
authors.initials: A
authors.lastname: Erler
authors.forename: Michael
authors.initials: M
authors.lastname: Hauser
authors.forename: Claudia
authors.initials: C
authors.lastname: Asam
authors.forename: Sonja
authors.initials: S
authors.lastname: Mutschlechner
authors.forename: Christof
authors.initials: C
authors.lastname: Ebner
authors.forename: Peter
authors.initials: P
authors.lastname: Briza
authors.forename: Fatima
authors.initials: F
authors.lastname: Ferreira
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D029662:Betula / Q000276:immunology*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D008024:Ligands; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification; D011058:Pollen / Q000276:immunology*; D011489:Protein Denaturation; D017510:Protein Folding; D020673:Protein Renaturation; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility
chemical_list: D000485:Allergens; D008024:Ligands; D010940:Plant Proteins; D011994:Recombinant Proteins; D007073:Immunoglobulin E