Mass Spectrometry & Proteomics

The introduction of two new ionization methods in mass spectrometry, electrospray and matrix-assisted laser desorption ionization (Maldi), in the late 80's have revolutionized protein chemistry. They both allow the accurate molecular weight determination of proteins and peptides, and from other biomolecules like nucleotides, sugars, etc... The inventors were granted the Nobel prize in chemistry in 2002. Further refining and continuous improvement of the equipment in terms of sensitivity and resolution, paved the way for mass spectrometry based proteomics, a key pillar in the current efforts for understanding Systems Biology.

We were happy to introduce the methods in Belgium in the early 1990’s. Thanks to financial support from Ghent University, FWO-Vlaanderen and the Hercules foundation we could continuously achieve state-of-the art equipment which are used in both quantitative proteomics, general protein chemistry and structural biology projects.

1. Electrospray Ionisation Mass Spectrometers

- A novel Waters SYNAPT HDMS mass spectrometer has been introduced in the lab mid 2009. This instrument combines the successful Quadrupole-Time-of-Flight configuration with a traveling wave ion mobility analyzer. This is instrument can be used both for LC-MS analysis of complex peptide samples and for intact protein and protein complex measurements. It is equipped with a Waters nanoAcquity UPLC system.

- A Q-TRAP 4000 linear ion trap instrument (Applied Biosystems). This instrument is a remarkable hybrid between a triple quadrupole and an ion trap instrument. The triple quad allows the typical scan functions (parent ion scans, MRM,) that are unique for this configuration, while Q3 is functioning as a linear ion trap with larger ion storage capacity, with MS3 functionality). It has an on-line nano or micro-capillary LC-system (Ultimate 3000 - RSLC system, Dionex).

2. Matrix-assisted Laser Desorption Mass Spectrometer

Maldi uses laserlight to desorb proteins and peptides from a matrix (existing of aromatic acid compounds). It allows proteins up to 500 kDa to be analyzed with 0.1% accuracy.

We have introduced a novel 4800 Proteomics analyzer (Applied Biosystems, spring 2008). This is a novel MALDI TOF/TOF instrument capable of high throughput protein identification in proteomics approaches, based on both peptide fingerprinting and MS/MS.

3. LTQ-FTICR-MS

The research group is a partner of the Gent University consortium operating an LTQ-FTICR MS system (Thermo) equipped with nanoLC (Agilent), UPLC (Thermo) and an Advion Triversa inlet system. This instrument is installed at the VIB department of Plant Systems Biology in the FVMS building at Ardoyen Research Park.

4. Proteomics

Our laboratory is working on proteomic analysis by both gel and non-gel based multidimensional separation technology. For gelelectrophoresis, we are equiped with a Bio-Rad platform based on a Dodecacell and PDQuest imaging, and analysis software. For non-gel applications we are using ITRAQ, SILAC and other stable isotopic labeling techniques combined with 2D-LC based on the LC Packings dual gradient 2D-LC system.