Depleted Prefractionated Human Plasma for Data Independent Acquisition Spectral Library

We established a spectral library for data-independent analysis of human plasma samples, using a pooled plasma sample from Danish adults. The samples were prepared for proteome analysis using an in-house developed protocol consuming one microliter of plasma per sample, and the tryptic digest and subsequent C18 purification was performed in 96-well plates. The spectral library consists of plasma which was analyzed neat (i.e. non-treated), and top 14 depleted. The samples were subjected to high pH reversed-phase peptide fractionated using C18 tips (8 fraction) and a HPLC system (20 fraction). The resulting peptides were analyzed on a Q Exactive with a capflow reversed-phase C18 LC-MS/MS setup in data-dependent mode. The final spectral library contains 21788 precursors and 1000 proteins (both at FDR<0.01).