Wai Tuck SohMaxime Le MignonNarissara SuratannonPattraporn SatitsuksanoaPantipa ChatchateeJongkonnee WongpiyaboronMukda VangveravongTicha RerkpattanapipatAtik SangasapaviliyaEmmanuel NonySurapon PiboonpocanunKiat RuxrungthamAlain JacquetPattarawat ThantiworasitPinya PulsawatTassalalpa DaengsuwanWannada LaisuanMalinee TongdeeNizchapha DchapaphapeaktakTadech BoonpiyathadChulalongkorn UniversityDivision of Allergy and ImmunologyQueen Sirikit National Institute of Child HealthMahidol UniversityPhramongkutklao College of MedicineStallergenesKing Chulalongkorn Memorial Hospital, Faculty of Medicine Chulalongkorn UniversityChildren HospitalFaculty of Medicine, Ramathibodi Hospital, Mahidol University2018-12-112019-03-142018-12-112019-03-142016-03-01International Archives of Allergy and Immunology. Vol.168, No.3 (2016), 150-16014230097101824382-s2.0-84973503067https://repository.li.mahidol.ac.th/handle/20.500.14594/40867© 2016 S. Karger AG, Basel. Background: The in-depth characterization of the recently identified house dust mite (HDM) major allergen Der p 23 requires the production of its recombinant counterpart because the natural allergen is poorly extractable from fecal pellets. This study aimed to provide a detailed physico-chemical characterization of recombinant Der p 23 (rDer p 23) as well as to investigate its IgE reactivity in a cohort of HDM-allergic patients from Thailand. Methods: Purified rDer p 23, secreted from recombinant Pichia pastoris, was characterized by mass spectrometry and circular dichroism analyses as well as for its chitin-binding activity. The IgE-binding frequency and allergenicity of Der p 23 were determined by ELISA and RBL-SX38 degranulation assays, respectively. Results: Purified intact rDer p 23 carried O-mannosylation and mainly adopted a random coil structure. Polyclonal antibodies to rDer p 23 can detect the corresponding natural allergen (nDer p 23) in aqueous fecal pellet extracts, suggesting that both forms of Der p 23 share common B-cell epitopes. Despite its homologies with chitin-binding proteins, both natural Der p 23 and rDer p 23 were unable to interact in vitro with chitin matrices. Of 222 Thai HDM-allergic patients tested, 54% displayed Der p 23-specific IgE responses. Finally, the allergenicity of rDer p 23 was confirmed by the degranulation of rat basophil leukemia cells. Conclusion: Our findings highlighted important levels of Der p 23 sensitizations in Thailand. Our study clearly suggested that rDer p 23 is likely more appropriate for HDM allergy component-resolved diagnosis than HDM extracts.Mahidol UniversityImmunology and MicrobiologyArticleSCOPUS10.1159/000442176