MicroSPE-nanoLC-ESI-MS/MS using 10-microm-i.d. silica-based monolithic columns for proteomics.
Silica-based monolithic capillary columns (25 cm x 10 microm 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 approximately 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.
Luo
Quanzhou
Q
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Page
Jason S
JS
Tang
Keqi
K
Smith
Richard D
RD
D002853:Chromatography, Liquid / Q000295:instrumentation*; D010455:Peptides / Q000032:analysis; D040901:Proteomics / Q000295:instrumentation*; D015203:Reproducibility of Results; D020592:Shewanella / Q000737:chemistry / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*; D014357:Trypsin / Q000737:chemistry
D010455:Peptides; D012822:Silicon Dioxide; D014357:Trypsin