Small-Scale Serial Size Exclusion Chromatography (s³SEC) for High Sensitivity Top-Down Proteomics of Large Proteoforms.

ABSTRACT: Top-down-mass spectrometry (MS)-based proteomics has emerged as a premier technology to examine proteins at the proteoform level, enabling characterization of genetic mutations, alternative splicing, and post-translational modifications. However, significant challenges that remain in top-down proteomics include the analysis of large proteoforms and the sensitivity required to examine proteoforms from minimal amounts of sample. To address these challenges, we have developed a new method termed "small-scale serial Size Exclusion Chromatography" (s³SEC), which incorporates a small-scale protein extraction (1 mg of tissue) and serial SEC without postfractionation sample handling, coupled with online high sensitivity capillary reversed-phase

SUBMITTER: Rogers HT 

PROVIDER: S-EPMC11298573 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature