Document Detail

Production and proteomic characterization of pharmaceutical-grade Dermatophagoides pteronyssinus and Dermatophagoides farinae extracts for allergy vaccines.
MedLine Citation:
PMID:  16741365     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: House dust mites (HDM) such as Dermatophagoides pteronyssinus and Dermatophagoides farinae represent a major cause of type 1 allergies worldwide. Hence large quantities of well-characterized HDM extracts are needed to prepare pharmaceutical-grade allergy vaccines. To this aim, the present study was undertaken to define optimal conditions for large-scale cultures. METHODS: D. pteronyssinus and D. farinae were grown on different media combining various proportions of wheat germ, yeast and synthetic amino acids (the latter resembling the composition of the human stratum corneum). Extracts thus obtained were analyzed for their total allergenic activity, as well as major allergen and protein contents, using immunosorbent assays, HPLC, immunoblotting, two-dimensional electrophoresis and peptide mass fingerprinting. RESULTS: An optimal culture medium (Stalmite APF) based on wheat germ, yeast and amino acids in defined proportion (42, 42 and 15% w/w, respectively) was selected to grow various HDM species with high yields. A detailed proteomic analysis revealed that D. pteronyssinus extracts generated under such conditions did not contain allergens originating from culture medium components and that major prevalent HDM allergens (i.e. groups 1, 2, 7, 10, 13 and 20) are found among the most abundant proteins in the D. pteronyssinus extract. Semiquantitative dot-blot assays confirmed the presence of Der p 3-10 as well as Der p 13 and 14 allergens within the extracts. CONCLUSIONS: We developed a well-defined medium allowing to grow various HDM species at an industrial scale in a highly reproducible manner. Extracts from mites produced under such pharmaceutical conditions contain all the relevant allergens for desensitization purposes and in vivo diagnosis.
Thierry Batard; Alexandre Hrabina; Xue Zhi Bi; Henri Chabre; Pierrick Lemoine; Marie-Noëlle Couret; Dominique Faccenda; Bertrand Villet; Philippe Harzic; Françoise André; Sock Yong Goh; Claude André; Fook Tim Chew; Philippe Moingeon
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Publication Detail:
Type:  Journal Article     Date:  2006-06-01
Journal Detail:
Title:  International archives of allergy and immunology     Volume:  140     ISSN:  1018-2438     ISO Abbreviation:  Int. Arch. Allergy Immunol.     Publication Date:  2006  
Date Detail:
Created Date:  2006-07-19     Completed Date:  2006-09-05     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9211652     Medline TA:  Int Arch Allergy Immunol     Country:  Switzerland    
Other Details:
Languages:  eng     Pagination:  295-305     Citation Subset:  IM    
Copyright Information:
Copyright 2006 S. Karger AG, Basel.
Recherche et Développement, Stallergènes, Antony, France.
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MeSH Terms
Antigens, Dermatophagoides / analysis,  isolation & purification*
Chromatography, High Pressure Liquid
Dermatophagoides farinae / chemistry*
Dermatophagoides pteronyssinus / chemistry*
Electrophoresis, Gel, Two-Dimensional
Hypersensitivity / drug therapy*,  immunology
Proteomics / methods
Radioallergosorbent Test
Triticum / immunology
Vaccines / immunology,  isolation & purification*,  therapeutic use
Yeasts / immunology
Reg. No./Substance:
0/Antigens, Dermatophagoides; 0/Dermatophagoides pteronyssinus antigen p 2; 0/Vaccines; EC 3.4.22.-/Dermatophagoides pteronyssinus antigen p 1

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