Influence of different sample preparation approaches on proteoform identification by top-down proteomics.

ABSTRACT: Top-down proteomics using mass spectrometry facilitates the identification of intact proteoforms, that is, all molecular forms of proteins. Multiple past advances have lead to the development of numerous sample preparation workflows. Here we systematically investigated the influence of different sample preparation steps on proteoform and protein identifications, including cell lysis, reduction and alkylation, proteoform enrichment, purification and fractionation. We found that all steps in sample preparation influence the subset of proteoforms identified (for example, their number, confidence, physicochemical properties and artificially generated modifications). The various sample preparation strategies resulted in complementary identifications, substantially increasing the proteome covera

SUBMITTER: Kaulich PT 

PROVIDER: S-EPMC11621018 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature