Liquid Chromatography

Liquid Chromatography

$We have a variety of liquid chromatography instruments, both online to our mass spectrometers and offline for peptide and protein fractionation.

 

Online to the Mass Spectrometers

Waters Acquity UPLC

Waters Acquity

Up front and feeding the 5600 and the QStar Elite are Waters nanoAcquity UPLCs. These ultra high pressure liquid chromatography instruments allow high resolution (~15 second base peak width) separation of peptides.

Dionex U3000 RSLC Nano

In order to take advantage of the speed and sensitivity of the LTQ Orbitrap Velos (Thermo) the ultra-high pressure liquid chromatography (UPLC) system; the RSLC U3000 (Dionex) was purchased to fractionate complex peptide mixtures online. The U3000 RSLC Nano has the ability to fractionate peptides with a base peak width of less than 12 seconds. The LTQ Orbitrap Velos and U3000 RSLC Nano set up allows the identification of low abundance peptides. Increasing the separation power of the liquid chromatography increases the quantity and quality of the peptides identified.

Dionex Ultimate

Dionex ultimate online to QTrap

Feeding the 4000 QTRAP is a Dionex Ultimate with switchos and Famos autosampler. Suitable fractionation of peptides is achieved using this instrument to allow identification of peptides using MRM methodology in the femto molar range.

Dionex U3000 and probot plate spotter

Dionex U3000

The U3000 is connected to a plate spotter so that complex peptide samples can be fractionated and spotted onto a MALDI plate (1664 spots per plate).

Offline

Dionex Ultimate

Dionex ultimate offline

This HPLC is used for sample fractionation offline from any mass spectrometers. Generally the instrument is used to fractionate complex peptide mixtures using either strong cation exchange (SCX) or high pH reverse phase chromatography. Each of these fractions is then loaded onto one of the LCs upfront of the mass spectrometers. This instrument is also used for protein fractionation, currently using monolithic or mixed bed (WCX and WAX) columns.

Agilent 1260 Infinity HPLC

Agilent 1260

The Agilent HPLC is used for depletion of clinical material, principally plasma however it is also capable of depleting other samples such as cerebral spinal fluid. This system depletes the top 14 proteins identified in plasma including Albumin and immunoglobulin G. The identification and quantitation of lower abundance proteins are therefore more accessible from the depleted sample.