Towards comprehensive plasma proteomics by orthogonal protease digestion

ABSTRACT: Rapid and consistent protein identification across large clinical cohorts is an important goal for clinical proteomics. With the development of data-independent technologies (DIA/SWATH-MS), it is now possible to analyze hundreds of samples with great reproducibility and quantitative accuracy. However, this technology benefits from empirically derived spectral libraries that define the detectable set of peptides and proteins. Here we apply a simple and accessible tip-based workflow for the generation of spectral libraries to provide a comprehensive overview on the plasma proteome in individuals with and without active tuberculosis (TB). To boost protein coverage, we utilized non-conventional proteases such as GluC and AspN together with the gold standard trypsin, identifying more than 30,000 peptides mapping to 3,309 proteins. Application of this library to quantify plasma proteome differences in TB infection recovered more than 400 proteins in 50 minutes of MS-acquisition, including diagnostic Mycobacterium Tuberculosis(Mtb) proteins that have previously been detectable primarily by antibody-based assays and intracellular proteins not previously described to be in plasma.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human) Mycobacterium Tuberculosis Tkk-01-0032

TISSUE(S): Blood Plasma

DISEASE(S): Pulmonary Tuberculosis

SUBMITTER: Andrea Fossati  

LAB HEAD: Danielle Swaney

PROVIDER: PXD025671 | Pride | 2021-07-29

REPOSITORIES: Pride