Online μSEC²-nRPLC-MS for Improved Sensitivity of Intact Protein Detection of IEF-Separated Nonhuman Primate Cerebrospinal Fluid Proteins.

ABSTRACT: Proteoform-resolved information, obtained by top-down (TD) "intact protein" proteomics, is expected to contribute substantially to the understanding of molecular pathogenic mechanisms and, in turn, identify novel therapeutic and diagnostic targets. However, the robustness of mass spectrometry (MS) analysis of intact proteins in complex biological samples is hindered by the high dynamic range in protein concentration and mass, protein instability, and buffer complexity. Here, we describe an evolutionary step for intact protein investigations through the online implementation of tandem microflow size-exclusion chromatography with nanoflow reversed-phase liquid chromatography and MS (μSEC²-nRPLC-MS). Online serial high-/low-pass SEC filtration overcomes the aforementioned hurdles t

SUBMITTER: Cline EN 

PROVIDER: S-EPMC10476446 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature