Efficient separations of intact proteins using slip-flow with nano-liquid chromatography-mass spectrometry.

ABSTRACT: A capillary with a pulled tip, densely packed with silica particles of 0.47 μm in diameter, is shown to provide higher peak capacity and sensitivity in the separation of intact proteins by reversed-phase liquid chromatography-mass spectrometry (LC-MS). For a C18 bonded phase, slip flow gave a 10-fold flow enhancement to allow for stable nanospray with a 4-cm column length. Model proteins were studied: ribonuclease A, trypsin inhibitor, and carbonic anhydrase, where the latter had impurities of superoxide dismutase and ubiquitin. The proteins were well separated at room temperature with negligible peak tailing. The peak capacity for ubiquitin was 195 for a 10-min gradient and 315 for a 40-min gradient based on Gaussian fitting of the entire peak, rather than extrapolating the full-width at

SUBMITTER: Wu Z 

PROVIDER: S-EPMC3982985 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature