Spin column-based peptide fractionation alternatives for streamlined tandem mass tag (SL-TMT) sample processing.

ABSTRACT: Fractionation is essential to achieving deep proteome coverage for sample multiplexing experiments where currently up to 18 samples can be analyzed concurrently. However, peptide fractionation (i.e., upstream of LC-MS/MS analysis) with a liquid chromatography system constrains sample processing as only a single sample can be fractionated at once. Here, we highlight the use of spin column-based methods which permit multiple multiplexed samples to be fractionated simultaneously. These methods require only a centrifuge and eliminate the need for a dedicated liquid chromatography system. We investigate peptide fractionation with strong anion exchange (SAX) and high-pH reversed phase (HPRP) spin columns, as well as a combination of both. In two separate experiments, we acquired deep proteome co

SUBMITTER: Liu X 

PROVIDER: S-EPMC9990130 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature