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

Result code in 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>

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

30304 - Public and environmental health

Project

<a href="/en/project/OC10019" target="_blank" >OC10019: Chemical biology with inhibotrs of digestive enzymes of mites: Searching of tools useful in suppression, detection and chemical biology of mites Acari: Acaridida</a><br>

Continuities

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

Publication year

2018

Confidentiality

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

Name of the periodical

Allergologia et immunopathologia

ISSN

0301-0546

e-ISSN

1578-1267

Volume of the periodical

46

Issue of the periodical within the volume

3

Country of publishing house

ES - SPAIN

Number of pages

8

Pages from-to

218-225

UT code for WoS article

000430786300003

EID of the result in the Scopus database

2-s2.0-85034587699