As a pooled donor blood product, cryoprecipitate carries higher risks of pathogen transmission. Pathogen inactivation (PI) improves the safety of cryoprecipitate, but its effects on hemostatic properties remain unclear. This study investigated protein expression in samples of pathogen inactivated cryoprecipitate (PI-cryo) using non-targeted quantitative proteomics. Specifically, cryoprecipitate samples were reduced and alkylated and digested with trypsin. Approximately 50 micrograms of tryptic digests were analyzed by microflowLC-MS/MS. The LC separation used a Waters ACQUITY UPLC and a 1 mm x 150 mm 1.7 micron CSH C18 column using a flow rate of 100 microliters/min, a column temperature of 55C and a gradient of 3-28% (v/v) MeCN:H2O containing 0.1% formic acid over 60 minutes. The LC was interfaced to a Thermo Q-Exactive HF-X using the HESI-II source. The MS analysis used data-independent acquisition (DIA)-MS with a 120,000 resolution precursor ion (MS1) scan from 375-1500 m/z, AGC target of 3E6 and maximum injection time (IT) of 20 ms. DIA-MS/MS was performed using 30,000 resolution, AGC target of 3e6 and maximum IT of 60 ms, and 27 V NCE. The DIA windows included 15 x 37 m/z windows spanning 400-941 m/z with 1 m/z overlaps; as well as 3 x 87 m/z windows spanning 941-1200 m/z. Data was analyzed using Spectronaut Pulsar X
[doi:10.25345/C5CH84]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: plasma ; apheresis ; cryoprecipiate ; microflow ; data-independent acquisition
Principal Investigators: (in alphabetical order) |
Matthew W. Foster, Duke University, USA Reed Kamyszek, University of Michigan, USA |
Submitting User: | mwfoster |
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