CEEPC/IPM/CMSC - Abstrakt prezentace

(Česká konference hmotnostní spektrometrie 2019 - ThS-009)
Proteomic analysis of exhaled breath condensates

Jana Václavková 1 *, Tatiana Gvozdiaková 2, Jana Vrbková 1, Petr Džubák 1, Dušan Holub 1, František Kopřiva 2, Vendula Látalová 2, Juraj Kultan 3, Vítězslav Kolek 3, Marián Hajdúch 1

  1. Institute of Molecular a Translational Medicine Olomouc
  2. Department of Pediatrics, University Hospital Olomouc
  3. Department of Respiratory Medicine, University Hospital Olomouc

Abstrakt

Exhaled breath condensate (EBC) represents a source of biomarkers such as proteins, arachidonic acid metabolites, vasoactive peptides amines, DNA, RNA, microRNA and small molecules. These biomarkers can provide valuable information about respiratory as well as systemic diseases. Finding non-invasive methods for early detection of lung diseases would be highly beneficial. Proteomic analysis of EBC has the potential to detect early changes in the status of the respiratory system. It could complement some invasive sampling methods in future and provide non-invasive lung diseases screening technique. Our study is focused on children’s asthma. The aim of this study is to identify biomarkers of asthma bronchiale in children’s exhaled breath condensate.
Exhaled breath condensate proteins in the sample are solubilized and denatured, reduced by DTT, digested by trypsin and concentration of peptides is measured. Samples are purified and diluted for HPLC/MS analysis which is performed in three technical replicates using high resolution LTQ Orbitrap Elite mass spectrometer (Thermo Scientific). Measured spectra are analyzed by Proteome Discoverer software (Thermo Scientific). Data are further statistically evaluated by Statistica and Bioconductor R – package.
In our study we collected and analyzed samples from 62 pediatric asthma patients. We have successfully analyzed the exhaled breath condensate of diagnosed and treated asthma and compared them with the samples of 62 healthy controls. We have found proteomic biomarkers of asthma and asthma treatment prediction. Discovered biomarkers will be validated by MRM mass spectrometry analysis and immunochemical methods.

* Korespondující autor: vaclavek.jana@centrum.cz

Poděkování:

This work is supported by the grants of AZV 16-32302A, AZV 16-32318A and by the National Sustainability Program (LO1304).


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