It was hypothesized that the severe inflammatory liver injury caused by alcoholic hepatitis (AH) would yield a unique peptidome profile in human patient plasma, and degraded fragments (degradome) of hepatic and extracelluar proteomes would change between AH patient groups defined by MELD scores. Following addition of iRT standards (Biognosys), the low molecular weight proteome (peptidome) was isolated from 100uL patient K3EDTA plasma using TCA precipitation, cleaned up by solid phase extraction on Waters Oasis HLB 96-well plates, and peptide concentrations estimated by A205nm on a Nanodrop. Peptides were separated by nanoUPLC on Proxeon 1000 thermostated at 50oC prior to nanoelectrospray into an Orbitrap ELITE. High resolution MS1 (240,000) were collected in the FT and MS2 were collected by ITMS. Data were analyzed by PEAKS Studio for de novo database analysis and PTM imputation. TIC-normalized XIC data were used for statistical analyses.
[doi:10.25345/C56688W0F]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: de novo sequencing ; biomarker ; degradome ; alcoholic hepatitis
Principal Investigators: (in alphabetical order) |
Michael L. Merchant, PhD, University of Louisville, USA |
Submitting User: | mmerchant |
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Owner | Reanalyses | |
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