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HIGHLY SELECTIVE ISOLATION OF TRYPTIC PHOSPHOPEPTIDES FROM THE JURKAT T-CELL LYSATE BY NOVEL NANOMATERIAL, E.G. TIO2 NANOTUBES DECORATED BY FE3O4 NANOPARTICLES

Rok: 2017

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

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Jazyk	Název	Abstrakt	Klíčová slova
eng	HIGHLY SELECTIVE ISOLATION OF TRYPTIC PHOSPHOPEPTIDES FROM THE JURKAT T-CELL LYSATE BY NOVEL NANOMATERIAL, E.G. TIO2 NANOTUBES DECORATED BY FE3O4 NANOPARTICLES	One of the challenges in phosphoproteomics is the enrichment of (multi)phosphorylated peptides as well as reduction of number of contaminating non-phosphorylated peptides to the lowest level. Techniques employed to phosphopeptide isolation utilize either metal ions (e.g. Fe3+) or metal oxides (e.g. TiO2), (1). TiO2-based nanomaterials widely applied for phoshphopeptides enrichment are not enough selective and elution efficiency for multiphoshorylated peptides is challenged. The aim of our work is to demonstrate the advantages of magnetically active nanomaterial based on TiO2 with nanotubular structure (TiO2NTs@Fe3O4NPs) compared to common TiO2 materials available today on the market. A peptide mixture consisted of tryptically digested phosphorylated and nonphosphorylated proteins was used as sample. The phosphopeptide enrichment was performed either with TiO2 microparticles (Titansphere, GL Sciences) or with the novel carrier TiO2NTs@Fe3O4NPs. All samples were analyzed by MALDI LTQ Orbitrap XL mass spectrometer and by LC-MS/MS – UltiMate 3000 RSLC-nanoHPLC system and Q Exactive mass spectrometer (all Thermo Scientific). Number of identified monophosphorylated peptides was similar for all tested materials. However, elution fractions obtained from the new magnetic nanomaterial contained higher portion of multiphosphorylated peptides, as compared to commonly used commercial TiO2 NPs. These results indicate higher rate of eluted and by MS identified phosphorylated peptides. Above that the contribution of non-phosphorylated peptides is significantly reduced. The presented TiO2NTs@Fe3O4NPs showed the high enrichment efficiency for multiphosphorylated peptides and simultaneously the elution fractions are not ―contaminated‖ by non-phosphorylated peptides. The benefit of this material is its magnetism, through which the nanomaterial can be well manipulated and effectively separated from aqueous phase.	PHOSPHOPEPTIDES; TIO2 NANOTUBES; FE3O4 NANOPARTICLES

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