Faculty of Biological Sciences

Instrumentation

The Facility has six electrospray mass spectrometers which are used in a variety of service and research applications. There is also an HDX manager as well as an excimer laser that is used for photochemical oxidation of proteins..

Synapt G2-Si

G2Si

The G2-Si is the most advaced instrument in the Facility and is used for a variety of applications. Electron transfer dissociation (ETD) technology enable top-down sequencing of proteins and large peptides. Coupled with an ACQUITY M-Class UPLC enables proteomics applications focussing on the structural chemical foot printing approaches. The IMS cell enable separation of analytes based on cross sectional area prior to mass analysis. Useful when dealing with extremely complex peptide mixtures adding an extra dimension of resolution. The IMS capabilties are also used for collision cross sectional are measurments of proteins and their complexes.

Xevo Q-ToF G2-XS

Xevo QTOF G2-XS This quadrupole orthogonal time of flight instrument has a segmented quadrupole collision cell. Coupled with the step wave ion guide makes this the most sensitive instrument in the Facility.In combination with a second ACQUITY M-Class UPLC this instrument is used for intact protein LC-MS as well as protein identification/mapping.

HDX Manager

HDX Using a refridgerated column compartment, this instrument enables online pepsin digestion of deuterium labelled proteins and subsequent reversed phase separation of proteolytic peptides. Coupled with mass spectrometry using the Synapt G2-Si instrument and Dynamx data processing software, the kinetics of solvent accessibiltiy of proteins under different conditions can be probed.

Excimer laser

Excminer laser The Coherent laser is used by the Facility to perform Fast Photochemical Oxidation of Proteins (FPOP). Proteins in solution with hydrogen peroxide are exposed to laser irradiation. This causes photolysis of the hydrogen peroxide. The resulting hydroxyl radicals react with the protein causing oxidation of solvent-exposed amino acid side chains. The sites of modification are mapped using LC-MS/MS.

Synapt G2-S

G2S This development instrument has been used for a variety of projects including IMS-MS analysis of oligonucleotide-ligand complexes and gas-phase hydrogen-deuterium exchange using deuterated ammonia for probing conformational change in proteins and their complexes.

Synapt HDMS

Synapt HDMS with 32k quadrupole The Synapt HDMS (Waters) system was purchased with funding from a BBSER REI grant in 2007. It is a quadrupole orthogonal time of flight mass spectrometer and also has a travelling wave ion mobility spectrometrydevice. The quadrupole has a mass range up to m/z 32 000 to enable MS/MS analysis of macromolecular biomolecules and non-covalent complexes. It has a NanoMate Triversa (Advion Biosciences) automated injection system and nanoESI interface.

LCT Premier

LCT Premier The LCT Premier (Waters) was purchased in 2004 with a Wellcome Trust grant and further financial support from the University of Leeds. It has been specially modified for the analysis of non-covalently bound macromolecular complexes and has the highest m/z range available commercially (m/z 60,000). It has a NanoMate (Advion Biosciences) automated injection system and a high field asymmetric waveform ion mobility spectrometry (FAIMS) device.

Synapt HDMS system

liquid chromatography-Synapt HDMS system The second Synapt HDMS (Waters) was purchased with SRIF4 funding in 2009. This instrument has a m/z 4000 quadrupole and is coupled online to an Ultimate 3000 HPLC system (Dionex). This instrument is used for the majority of the service analyses. The LC-MS/MS capabilities (in conjuction with ProteinLynx Global Server software purchased with HEFCE funds in 2010) allow protein identification and post translational modification site determination.