Web Release Date: December 15,
MicroSPE-nanoLC-ESI-MS/MS Using 10-
m-i.d.
Silica-Based Monolithic Columns for Proteomics
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352
Received for review August 28, 2006. Accepted October 31, 2006.
Abstract:
Silica-based monolithic capillary columns (25 cm × 10
m i.d.) with integrated nanoESI emitters have been
developed to provide high-quality and robust microSPE-nanoLC-ESI-MS analyses. The integrated nanoESI emitter
adds no dead volume to the LC separation, allowing stable
electrospray operation at flow rates of ~10 nL/min. In
an initial application with a linear ion trap MS, we
identified 5510 unique peptides that covered 1443
distinct Shewanella oneidensis proteins from a 300-ng
tryptic digest sample in a single 4-h LC-MS/MS analysis.
The use of an integrated monolithic ESI emitter provided
enhanced resistance to clogging and provided good run-to-run reproducibility.
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