Non-protease native allergens partially purified from bodies of eight domestic mites using p-aminobenzamidine ligand

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027006%3A_____%2F18%3A00004456" target="_blank" >RIV/00027006:_____/18:00004456 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.aller.2017.07.004" target="_blank" >http://dx.doi.org/10.1016/j.aller.2017.07.004</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.aller.2017.07.004" target="_blank" >10.1016/j.aller.2017.07.004</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Non-protease native allergens partially purified from bodies of eight domestic mites using p-aminobenzamidine ligand

Popis výsledku v původním jazyce

Background: Optimised purification steps for concentrating trace target native antigens are needed. Combining the p-aminobenzamidine ligand with protease inactivation enables partial purification of mite non-protease allergens lacking proteases. Objective: We sought to analyse in detail proteins obtained using this method from eight species of synanthropic acaridid mites and tested IgE reactivity using pooled human sera. Materials and methods: Proteins affinity bound to p-aminobenzamidine as a ligand were identifled by MALDI TOF/TOF. After electroblotting, the proteins were visualised using the fluorescent SYPRO-Ruby protein blot stain, and IgE reactivity was further analysed using pooled human sera collected from patients allergic to house dust mites. Results: MS/MS identification confirmed previous results that no proteases were purified. Protein patterns corresponding to the allergens Der f 7, Der f 30 and actins indicated that these proteins are purified using p-aminobenzamidine and are present across a wide spectrum of acaridid mites. When using Dermatophagoides farinae, apolipophorins (Der f 14), chitinase-like Der f 15 and 18, 70-kDa heat shock protein, and a Der f Alt a10 allergen homolog (gi137958173) were also detected. The target antigens tropomyosins and paramyosins showed similar IgE binding among the mite species tested. IgE reactivity with miscellaneous D. farinae antigen was also observed. Conclusions: Partial purification of mite non-protease antigens using a strategy combining paminobenzamidine with protease inactivation was verified by 1D-E and 2D-E analyses. IgE binding to p-aminobenzamidine-purified native non-protease mite antigens was tested using pooled sera. This preliminary study allows for further work on individual serum samples, allowing confirmation of immunoreactivity.

Název v anglickém jazyce

Non-protease native allergens partially purified from bodies of eight domestic mites using p-aminobenzamidine ligand

Popis výsledku anglicky

Background: Optimised purification steps for concentrating trace target native antigens are needed. Combining the p-aminobenzamidine ligand with protease inactivation enables partial purification of mite non-protease allergens lacking proteases. Objective: We sought to analyse in detail proteins obtained using this method from eight species of synanthropic acaridid mites and tested IgE reactivity using pooled human sera. Materials and methods: Proteins affinity bound to p-aminobenzamidine as a ligand were identifled by MALDI TOF/TOF. After electroblotting, the proteins were visualised using the fluorescent SYPRO-Ruby protein blot stain, and IgE reactivity was further analysed using pooled human sera collected from patients allergic to house dust mites. Results: MS/MS identification confirmed previous results that no proteases were purified. Protein patterns corresponding to the allergens Der f 7, Der f 30 and actins indicated that these proteins are purified using p-aminobenzamidine and are present across a wide spectrum of acaridid mites. When using Dermatophagoides farinae, apolipophorins (Der f 14), chitinase-like Der f 15 and 18, 70-kDa heat shock protein, and a Der f Alt a10 allergen homolog (gi137958173) were also detected. The target antigens tropomyosins and paramyosins showed similar IgE binding among the mite species tested. IgE reactivity with miscellaneous D. farinae antigen was also observed. Conclusions: Partial purification of mite non-protease antigens using a strategy combining paminobenzamidine with protease inactivation was verified by 1D-E and 2D-E analyses. IgE binding to p-aminobenzamidine-purified native non-protease mite antigens was tested using pooled sera. This preliminary study allows for further work on individual serum samples, allowing confirmation of immunoreactivity.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

30304 - Public and environmental health

Projekt

<a href="/cs/project/OC10019" target="_blank" >OC10019: Chemická biologie s inhibitory trávicích enzymů roztočů: Hledání nástrojů využitelných v supresi, detekci a chemické biologii roztočů Acari: Acaridida</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Allergologia et immunopathologia

ISSN

0301-0546

e-ISSN

1578-1267

Svazek periodika

46

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

ES - Španělské království

Počet stran výsledku

8

Strana od-do

218-225

Kód UT WoS článku

000430786300003

EID výsledku v databázi Scopus

2-s2.0-85034587699