*4.2. Sample Preparation*

The plasma samples were depleted from albumin and immunoglobulin using ProteoPrep depletion columns (Sigma-Aldrich, St. Louis, MO, USA) according to the manufacturer's protocol. Proteins from the urine samples were concentrated with Amicon Ultra-4 3 kDa centrifugal filter columns (Merck Millipore, County Cork, Ireland). The protein concentrations were determined by UV-1280 spectrophotometer (Shimadzu, Kyoto, Japan) and samples with 4 μg of protein were taken for mass spectrometry. The proteins were reduced by dithiothreitol, alkylated by iodoacetamide, and digested by trypsin according to the manufacturer's protocols (Thermo Scientific, Waltham, MA, USA). The samples were desalted by 10 μL C18 pipette tips from the same manufacturer in accordance with its protocol and dried before analysis.

#### *4.3. Liquid Chromatography and Mass Spectrometry*

The samples were dissolved in phase "A" (0.1% of formic acid) and 2 μg of each sample were injected into the Dionex UltiMate 3000 RSLC nano liquid chromatographer (Thermo Scientific, USA) with Acclaim RSLC PepMap C18 separation column (15 cm length, 75 μm inner diameter, 2 μm particles). The solvent gradient was increased from 0% to 40% of phase "B" (0.1% of formic acid in 80% acetonitrile) for 90 min, maintaining a constant flow rate of 200 nL/min. The Orbitrap Fusion mass spectrometer (Thermo Scientific, USA) was operated in data-dependent mode with scans of parent and fragment ions changing in cycle of 4 s. Full scans were made at a resolution of 60,000 by the Orbitrap mass detector, and fragments generated by high energy collision dissociation (HCD) were registered by ion trap at normal rate.
