MassIVE MSV000086867

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Protein profiles of extracellular vesicles from healthy preschool children born late preterm (LP) and full-term (T).

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Description

The isolated Evs from urine sample (from healthy preschool children born late preterm (LP) and full-term (T)) were resuspended in 0.1 M ammonium bicarbonate, pH 7.9 and were trypsinized according to procedures previously reported, using 50 microg of proteins from each sample. One microliter (about 1 micro g injected) of trypsin-digested mixtures was analyzed by nano-cromatography equipped with a cHiPLC-nanoflex system (Eksigent, AB SCIEX, USA) coupled to a Q-Exactive mass spectrometer (Thermo Fisher Scientific, USA), through a 65 min gradient of 5-45% of eluent B (eluent A, 0.1% formic acid in water; eluent B, 0.1% formic acid in acetonitrile), at a flow-rate of 300 nL/min. Full mass spectra were recorded in positive ion mode over a 400-1600 m/z range at a 70000 FWHM resolution, followed by 10 MS/MS spectra, at a resolution of 17,500 FWHM, generated in a data-dependent manner on the most abundant ions. All data generated were searched using Proteome Discoverer 2.1 platform (Thermoscientific) based on SEQUEST search engine and human protein database (70726 entries, downloaded on January 2017 from UNIPROT website, www.uniprot.gov). The obtained protein lists were aligned, normalized and then processed by means of Linear Discriminant Analysis (LDA). For assigning each subject to a specific group the alpha-value parameter was calculated according to the extracted marker proteins [doi:10.25345/C56796]

Keywords: Urine, extracellular vescicles, respiratory disease, children

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Principal Investigators:
(in alphabetical order) 		Dario Di Silvestre, Institute of Biomedical Technologies, Italy
Pierluigi Mauri, Institute of Biomedical Technologies, Italy
Submitting User: Dds100
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