January 15, 2007
Journal Article

MicroSPE-nanoLC-ESI-MS/MS using 10-µm-i.d. silica-based monolithic columns for proteomics

Abstract

Silica-based monolithic narrow bore capillary columns (25 cm ? 10 µm i.d.) with an integrated ESI emitter has been developed to provide high quality and robust microSPE-nanoLC-ESI-MS analyses. The integrated ESI emitter adds no dead volume to the LC separation, allowing stable electrospray performance to be obtained at flow rates of ?10 nL/min. In an initial application we identified 5510 unique peptides covering 1443 distinct Shewanella oneidensis proteins from a 300 ng tryptic digest sample in a single 4-h LC-MS/MS analysis using a linear ion trap MS (LTQ). We found the use of an integrated monolithic ESI emitter provided enhanced resistance to clogging and good run-to-run reproducibility, with an average variation of ~25% for triplicate analyses.

Revised: October 1, 2015 | Published: January 15, 2007

Citation

Luo Q., J.S. Page, K. Tang, and R.D. Smith. 2007. MicroSPE-nanoLC-ESI-MS/MS using 10-µm-i.d. silica-based monolithic columns for proteomics. Analytical Chemistry 79, no. 2:540-545. PNNL-SA-49661.