Figure 1. Integrated proteomics and N-glycoproteome analysis workflow applied to urine samples from bladder cancer patients and healthy subjects.
Following filtration with 3kDA filters, urinary proteins were concentrated and digested with trypsin. Each sample was labeled with 10-plex Tandem Mass Tags (TMT) labeling kit and pooled. One-tenth of the pooled sample was used for global proteomics experiment and remaining was enriched for N-glycoproteomic analysis. Samples were analyzed in Q Exactive HF-X Hybrid Quadrupole-Orbitrap mass spectrometer and the files were searched against Mascot and Sequest HT search engines. Data analysis was carried out using Perseus. exe 1.6.5.0.