On-line, Chip-Based Strategy for Increased Depth of Coverage in Proteomic Samples

摘要

Multi-dimensional chromatography is a key technology for working on complex proteomes and obtaining deeper depth of coverage of proteins. Using high pH reverse phase as the first dimension has been increasing in popularity because of its higher resolution and reduced need for salt. Here, the workflow was implemented using two cHiPLC systems for ease of use and fast switching between workflows. While further optimization can still be performed on the demonstrated workflow, substantial improvements in numbers of peptides identified was shown using the on-line 2D-LC RP-RP workflow compared to 1D separations.