Proteomic analysis of dried blood from the Sepsis Characterization in Kilimanjaro (SICK) study

ABSTRACT: Venous-EDTA whole blood was captured on 20 microliter Mitra devices at days 0,1,3,28 after hospital admission with Sepsis-2 criteria. Mitra tips were processed in Matrix tubes using deoxycholate-assisted in solution trypsin digestion. Approximately 1 mg of digests was enriched for N-glycopeptidesusing spin tips packed with Polyhydroxyethyl A followed by serial enrichment of phosphopeptides from the flow through using Ti-IMAC. LC-MS/MS of unenriched and phosphopeptide-enriched samples used microflow LC (30 samples-per-day) and Evosep LC (60 SPD), respectively, with data-dependent acquisition using an Orbitrap Astral MS. LC-MS/MS of N-glycopeptide-enriched samples used an Evosep LC (60 SPD) and either stepped-collision energy data-dependent acquisition (Exploris 480) or separate DDA and DIA with a 35% NCE using the Orbitrap Astral. Data analysis used Spectronaut and Glyco-Decipher.

INSTRUMENT(S): Orbitrap Astral, Orbitrap Exploris 480

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Matthew W. Foster  

PROVIDER: MSV000097000 | MassIVE | Thu Jan 30 11:58:00 GMT 2025

SECONDARY ACCESSION(S): PXD060377

REPOSITORIES: MassIVE