8 Allergenic Composition of Polymerized Allergen Extracts of Betula verrucosa, Dermatophagoides Pteronyssinus and Phleum Pratense
| Autor: | Miguel Casanovas, José Luis Subiza, Barbara Cases, Eva Abel Fernandez, Jose Ignacio Tudela, Enrique Fernández-Caldas |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Pulmonary and Respiratory Medicine
biology business.industry Oral Abstract Session Immunology respiratory system medicine.disease_cause biology.organism_classification Ige binding Abstracts of the XXII World Allergy Congress Qualitative composition respiratory tract diseases Phleum chemistry.chemical_compound Allergen Biochemistry chemistry immune system diseases otorhinolaryngologic diseases medicine Immunology and Allergy Composition (visual arts) Glutaraldehyde business |
| Zdroj: | The World Allergy Organization Journal |
| ISSN: | 1939-4551 |
| Popis: | Background Allergoids have been successfully used in the treatment of respiratory allergic diseases. They are modified allergen extracts that allow the administration of high allergen doses, due to their reduced IgE binding capacity.They maintain allergen-specific T-cell recognition. Since they are native allergen extracts that have been polymerized with glutaraldehyde, identification of the allergenic molecules requires more complicated methods. The aim of the study was to determine the qualitative composition of different polymerized extracts and investigate the presence of defined allergenic molecules using Mass spectrometry. Methods Proteomic analysis was carried out at the Proteomics Facility of the Hospital Nacional de Parapléjicos (Toledo, Spain). After reduction and alkylation, proteins were digested with trypsin and the resulting peptides were cleaned using C18 SpinTips Sample Prep Kit; peptides were separated on an Ultimate nano-LC system using a Monolithic C18 column in combination with a precolumn for salt removal. Fractionation of the peptides was performed with a Probot microfraction collector and MS and MS/MS analysis of offline spotted peptide samples were performed using the Applied Biosystems 4800 plus MALDI TOF/TOF Analyzer mass spectrometer. ProteinPilot Software V 2.0.1 and the Paragon algorithm were used for the identification of the proteins. Each MS/MS spectrum was searched against the SwissProt 2010_10 database, Uniprot-Viridiplantae database and Uniprot_Betula database. Results Analysis of the peptides revealed the presence of native allergens in the polymerized extracts: Der p 1, Der p 2, Der p 3, Der p 8 and Der p 11 in D. pteronyssinus; Bet v 2, Bet v 6, Bet v 7 and several Bet v 1 isoforms in B. verrucosa and Phl p 1, Phl p 3, Phl p 5, Phl p 11 and Phl p 12 in P. pratense allergoids. In all cases, potential allergenic proteins were also identified, including ubiquitin, actin, Eenolase, fructose-bisphosphate aldolase, luminal-binding protein (Heat shock protein 70), calmodulin, among others. Conclusions The characterization of the allergenic composition of allergoids is possible using MS/MS analysis. The analysis confirms the presence of native allergens in the allergoids. Mayor allergens are preserved during polymerization. |
| Databáze: | OpenAIRE |
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