Nanoflow low pressure high peak capacity single dimension LC-MS/MS platform for high-throughput, in-depth analysis of mammalian proteomes.

ABSTRACT: The use of narrow bore LC capillaries operated at ultralow flow rates coupled with mass spectrometry provides a desirable convergence of figures of merit to support high-performance LC-MS/MS analysis. This configuration provides a viable means to achieve in-depth protein sequence coverage while maintaining a high rate of data production. Here we explore potential performance improvements afforded by use of 25 ?m × 100 cm columns fabricated with 5 ?m diameter reversed phase particles and integrated electrospray emitter tips. These columns achieve a separation peak capacity of ?750 in a 600-min gradient, with average chromatographic peak widths of less than 1 min. At room temperature, a pressure drop of only ?1500 psi is sufficient to maintain an effluent flow rate of ?10 nL/min. Using mouse embryonic stem cells as a model for complex mammalian proteomes, we reproducibly identify over 4000 proteins across duplicate 600 min LC-MS/MS analyses.

SUBMITTER: Zhou F 

PROVIDER: S-EPMC3416051 | biostudies-literature | 2012 Jun

REPOSITORIES: biostudies-literature