Liquid Chromatography

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The Centre for Protein Research is equipped with different LC-systems suitable for the separation of peptides, proteins and various polar and non-polar compounds. We use a nanoflow/microflow Ultimate 3000 HPLC-instrument which is inline coupled to a Probot fraction collector for coupling RP-HPLC to MALDI tandem TOF-MS. Another Ultimate 3000 uHPLC system is inline coupled to the LTQ-Orbitrap MS. Both systems are equipped with dual pump systems for 1- and 2-dimensional liquid chromatography. In 2-D LC of peptides usually strong cation exchange liquid chromatography (SCX-LC) is connected upstream to nano-flow reversed phase liquid chromatography (RP-LC). An Eksigent ekspert 415 nanoflow uHPLC is inline coupled to 5600+ TripleTOF-MS for accurate retention time reproducibility in PRM/MRM and SWATH experiments.

Depending on the application and stationary phase we operate the nanoflow LC-systems at different flow rates between 200 and 800 nL/min. Different stationary phases are used for the separation of proteins (C4) as well as polar (HILIC, N2-HILIC, hypercarb) and non-polar compounds (C8, C18).