High-performance proteomics using nano-, capillary- and micro-flow chromatographic separations

In this study, we systematically evaluated the performance of μLC (50, 10 μL/min), capLC (5, and 1.5 μL/min), and nLC (0.3 μL/min ) flow-rates for proteome analysis using a Vanquish Neo HPLC coupled with a Q Exactive HF-X mass spectrometer. Serial dilutions of HeLa cell line digests were used for benchmarking the different setups and we intentionally fixed the total analysis time from injection-to-injection time of 60 minutes (24 samples per day). Showcase studies were also done on lable-free phospo-proteomics and kinobeads pull-down experiments.