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Plasma samples were depleted by a MARS Hu-14 column and subsequently concentrated by 3 kDa ultracentrifugation filters. Next, samples were reduced, cysteine blocked and trypsin digested before iTRAQ labeling. The iTRAQ labeled peptides were fractioned into 60 fractions using a mixed-mode reverse phase anion exchanger. Finally, fractions were analyzed on an LTQ-Orbitrap Velos Pro connected to a Dionex Ultimate NCR-3000RS LC system.
