title: Characterization of the house dust mite allergen Der p 21 produced in Pichia pastoris.
abstract: BACKGROUND
  The development of recombinant house dust mite (HDM) allergens opened the way for the in-depth characterization of these molecules but also provided new opportunities to refine the diagnostic procedures of HDM allergy as well as the allergen-specific immunotherapy through tailor-made treatments.
  
  
  OBJECTIVE
  In the present study, the HDM allergen Der p 21 was expressed in Pichia pastoris under a secreted form. The physico-chemical as well as the allergenic characterizations of recombinant Der p 21 (rDer p 21) were performed.
  
  
  METHODS
  Purified rDer p 21, secreted from recombinant P. pastoris was characterized by CD and MS analysis and the frequency of IgE reactivity was determined by ELISA using 96 sera of HDM-allergic patients from Bangkok. The direct airway epithelial cell activation by rDer p 21 was also evaluated.
  
  
  RESULTS
  rDer p 21 was highly expressed under a secreted form in P. pastoris. The physico-chemical characterization of purified rDer p 21 showed that the allergen displayed appropriate α-helix secondary structure content although a two amino acids truncation at the N-terminus of the protein was evidenced by MS. The prevalence of IgE reactivity to rDer p 21 reached 25% in the cohort of the HDM-allergic patients. rDer p 21 could trigger IL-8 production in airway epithelial cells through TLR2-dependent signaling.
  
  
  CONCLUSIONS
  Properly folded rDer p 21 produced in P. pastoris is appropriate for HDM allergy diagnosis as well for future recombinant allergen-based specific immunotherapy.
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Pinya
authors.initials: P
authors.lastname: Pulsawat
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Montalee
authors.initials: M
authors.lastname: Theeraapisakkun
authors.affiliation: Stallergenes, Antony 92160, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Nony
authors.affiliation: Stallergenes, Antony 92160, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Le Mignon
authors.affiliation: Stallergenes, Antony 92160, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Jain
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Arun
authors.initials: A
authors.lastname: Buaklin
authors.affiliation: Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Jongkonnee
authors.initials: J
authors.lastname: Wongpiyabovorn
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Kiat
authors.initials: K
authors.lastname: Ruxrungtham
authors.affiliation: Department of Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. Electronic address: alain.j@chula.ac.th.
authors.forename: Alain
authors.initials: A
authors.lastname: Jacquet
mesh_terms: D000485:Allergens / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D060829:Arthropod Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D003001:Cloning, Molecular; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D016209:Interleukin-8 / Q000096:biosynthesis; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D017433:Protein Structure, Secondary; D039981:Pyroglyphidae / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D020545:Respiratory Mucosa / Q000276:immunology; D015398:Signal Transduction / Q000276:immunology; D051195:Toll-Like Receptor 2 / Q000276:immunology
chemical_list: D000485:Allergens; D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; C508606:CXCL8 protein, human; D016209:Interleukin-8; D011994:Recombinant Proteins; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; D007073:Immunoglobulin E
keywords: Der p 21; House dust mite; P. pastoris; Recombinant