A newcomer's guide to nano-liquid-chromatography of peptides.
LC-MS/MS is one of the most powerful techniques in the field of proteomics allowing high throughput identification of proteins out of complex protein mixtures. Besides high sample throughput, the analytical sensitivity is one of the major benefits of this technology. A prerequisite for sensitive LC-MS/MS approaches is chromatography with very low flow rates in the nanoliter per minute range, usually referred to as nano-liquid chromatography (nano-LC). However, to perform this separation technology, an appropriate instrumental setup as well experienced operators are a prerequisite. The aim of this chapter is to help nano-LC newcomers to get introduced to this fascinating technology. Technical components of nano-LC systems like solvent delivery systems, sample injection systems, and nano-chromatography columns are described. Detailed procedures to mount, test, and operate the system are outlined, and advices for an effective troubleshooting are provided.
Fröhlich
Thomas
T
Laboratory for Functional Genome Analysis LAFUGA, Gene Center, Ludwig-Maximilians-University Munich, Feodor-Lynen-Str. 25, 81377 Munich, Germany.
Arnold
Georg J
GJ
D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D036103:Nanotechnology / Q000379:methods*; D010446:Peptide Fragments / Q000032:analysis* / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation* / Q000379:methods*
D010446:Peptide Fragments