Comparative Analysis of Proteoform Clean-Up Methods for In-Depth Top-Down Proteomics.

ABSTRACT: Proteoforms, the endogenous forms of proteins that take into account all sources of variation (genetic as well as co- and post-translational), may be more relevant for understanding complex biological mechanisms, including disease phenotypes, than generically defined "protein families". Mass spectrometry-based top-down proteomics often utilizes electrophoretic techniques that rely on sodium dodecyl sulfate (SDS) to fractionate proteoforms prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Methanol-chloroform-water (MCW) precipitation is commonly utilized for SDS removal due to its efficacy and low cost, yet it may lead to poor recovery of smaller proteoforms. As a technical contribution to top-down proteomics, four commercial SDS clean-up alternatives were benchma

SUBMITTER: Williams AA 

PROVIDER: S-EPMC12716119 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature