Optimizing Proximity Proteomics on the EvoSep-timsTOF LC-MS System.

ABSTRACT: Proximity-dependent biotinylation (BioID) is a powerful means of exploring the cellular environments in which proteins reside. Expressing a protein of interest (bait) fused to a biotin ligase and adding biotin induces the covalent biotinylation of proximal partners (preys), which are recovered on streptavidin beads and identified by MS. However, a major technical limitation of BioID is peptide carryover into subsequent MS runs. This is typically mitigated via lengthy intersample wash cycles, which lowers throughput considerably. The aim of this study was to optimize BioID sample acquisition using an EvoSep LC system coupled to a timsTOF mass spectrometer, which has higher throughput and sensitivity than our current system, with less carryover. Our efforts resulted in an ∼15-fold increase i

SUBMITTER: Kitaygorodsky J 

PROVIDER: S-EPMC12329395 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature