title: Allergenicity of native and recombinant major allergen groups 1 and 2 of Dermatophagoides mites in mite sensitive Thai patients.
abstract: OBJECTIVE
  Natural allergenic extracts using for diagnosis and immunotherapy may have batch-to-batch variations and contaminations with unrefined allergens or non-allergenic components. Thus, recombinant allergen is believed to overcome these shortcomings. In this study, native and recombinant allergens of group 1 and 2 of Dermatophagoides mites were produced and their allergenicities were compared.
  
  
  METHODS
  Native allergens were prepared by MAb affinity chromatography. All recombinant allergens were produced in E. coli expression system. IgE reactivities of these allergens were determined by IgE-ELISA.
  
  
  RESULTS
  The native and recombinant Der p 1, Der p 2, Der f 1, Der f 2 had molecular weights of approximately 25, 15, 25 and 15 kDa, respectively. IgE reactivities of nDer p 1, nDer f 1, rDer p 1 and rDer f 1 were 96.67%, 90%, 43.33% and 46.67%, respectively. Allergenicities of nDer p 2, nDer f 2, rDer p 2 and rDer f 2 were 86.67%, 96.43%, 76.67% and 89.29%, respectively. The findings indicated that recombinant group-1 products were minor allergens which revealed no correlation with their native forms. In contrast, recombinant group-2 allergens were major allergens and showed a significant correlation to their native allergens.
  
  
  CONCLUSIONS
  We successfully produced native and recombinant group-1 and group-2 allergens. According to their allergenicities, recombinant Der p 2 and rDer f 2 have potential to replace native allergen in diagnostic and therapeutic extracts. Moreover, they can employ as a standard reagent to measure the amount of group 2 allergen in the environment by sandwich-ELISA and utilise this as an immunogen for MAb production.
authors.affiliation: Office for Research and Development, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Nitat
authors.initials: N
authors.lastname: Sookrung
authors.forename: Jintarat
authors.initials: J
authors.lastname: Choopong
authors.forename: Watee
authors.initials: W
authors.lastname: Seesuay
authors.forename: Nitaya
authors.initials: N
authors.lastname: Indrawattana
authors.forename: Wanpen
authors.initials: W
authors.lastname: Chaicumpa
authors.forename: Anchalee
authors.initials: A
authors.lastname: Tungtrongchitr
mesh_terms: D000328:Adult; D000818:Animals; D039741:Antigens, Dermatophagoides / Q000276:immunology*; D060829:Arthropod Proteins / Q000276:immunology; D003546:Cysteine Endopeptidases / Q000276:immunology; D005260:Female; D006801:Humans; D006967:Hypersensitivity / Q000209:etiology*; D007073:Immunoglobulin E / Q000276:immunology; D008297:Male; D008875:Middle Aged; D039981:Pyroglyphidae / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D039741:Antigens, Dermatophagoides; D060829:Arthropod Proteins; C451160:Dermatophagoides farinae antigen f 2; C451163:Dermatophagoides pteronyssinus antigen p 2; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D003546:Cysteine Endopeptidases; C451159:Dermatophagoides farinae antigen f 1; C451162:Dermatophagoides pteronyssinus antigen p 1