Fully automated four-column capillary LC-MS system for maximizing throughput in proteomic analyses.
We describe a four-column, high-pressure capillary liquid chromatography (LC) system for robust, high-throughput liquid chromatography-mass spectrometry (LC-MS(/MS)) analyses. This system performs multiple LC separations in parallel, but staggers each of them such that the data-rich region of each separation is sampled sequentially. By allowing nearly continuous data acquisition, this design maximizes the use of the mass spectrometer. Each analytical column is connected to a corresponding ESI emitter in order to avoid the use of postcolumn switching and associated dead volume issues. Encoding translation stages are employed to sequentially position the emitters at the MS inlet. The high reproducibility of this system is demonstrated using consecutive analyses of global tryptic digest of the microbe Shewanella oneidensis.
Livesay
Eric A
EA
Biological Sciences Division, Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99354, USA.
Tang
Keqi
K
Taylor
Beverley K
BK
Buschbach
Michael A
MA
Hopkins
Derek F
DF
LaMarche
Brian L
BL
Zhao
Rui
R
Shen
Yufeng
Y
Orton
Daniel J
DJ
Moore
Ronald J
RJ
Kelly
Ryan T
RT
Udseth
Harold R
HR
Smith
Richard D
RD
D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation*; D013058:Mass Spectrometry / Q000295:instrumentation*; D010455:Peptides / Q000737:chemistry; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D020592:Shewanella / Q000201:enzymology; D014357:Trypsin / Q000378:metabolism
D010455:Peptides; D014357:Trypsin