ToF instruments

Those instruments are characterized by a fast scan rate while providing good resolution and mass precision. They are mainly used in our facility:

– To measure the mass of entire proteins and other macromolecules such as nucleic acids or polymers

– For protein identification of samples coming from 2D-gel separation (MALDI-TOF/TOF)

– And for molecular imaging of peptides and proteins.

Leader instruments

ES-Synapt G2 HDMS (Waters): this spectrometer has Ion Mobility capabilities allowing separating conformational isomers and can perform ETD fragmentation.

MALDI UltrafleXtreme TOF/TOF (Bruker): with its 2 kHz laser and its MS/MS capabilities, this instrument is indicated for high throughput MALDI applications such has protein identifications and molecular imaging of various samples such as biological tissues.

Other instruments available

ESI-Synapt G1 HDMS (Waters)

ESI-Q-ToF Ultima Global (Waters)

ESI-Micro ToF (Bruker)

FT instruments

Thanks to their high mass precision and high resolution, these instruments are essential for applications requiring sub-ppm mass determination or for very complex mixtures containing compounds with very close masses such as digested protein extracts or lipids molecular imaging.

Instruments available

Q Exactive Plus and Q Exactive (Thermo scientific) : these spectrometers are mainly used for differential proteomic purpose, as it allows identifying several thousand proteins from biological samples such as cytoplasmic extracts.

ESI/MALDI-FT-ICR MS SolariX 9.4T (Bruker) : this instrument is dedicated to exact mass measurement of small compounds for raw formula determination and structural elucidation and to molecular imaging of lipids and small molecules.

Ion Trap instrument

Our Ion trap instrument is an ESI-Ion trap AmaZon Speed ETD (Bruker) and is dedicated to protein identification in simple mixtures. It has ETD capability that allows localizing labile post translational modifications such as phosphorylation and glycosylation. This instrument works under FAMHP agreement and is qualified for batch characterization of proteins used as raw material in biotechnological/pharmaceutical industries.

Triple Quadrupole instruments

TQ instruments are dedicated to targeted quantitative analyses of peptides and small molecules. An ESI-Xevo TQ-S (Waters) is available that can be hyphenated with any of the UPLC systems.

An older instrument is still available: ESI-Quattro Ultima Pt (Waters).

HPLC/UPLC instruments

Separation is an essential step in the study of complex mixtures. For proteomic applications, both 1D and 2D are used in routine in the facility. Several chromatographic systems ranging from mL/min scale to the nL/min flow rate scale are available in the facility and can be coupled to the different mass spectrometers, depending on the application needs.

Two 2D NanoAcquity UPLC (Waters)

2D-nano-HPLC Ultimate 3000 (Dionex)

Two Acquity UPLC I-Class (UV detector available, Waters)

microUPLC M-class (Waters)

TriVersa NanoMate (Advion)

Sample preparation

Janus liquid handling workstation (Perkin Elmer) is dedicated to automated gel spot digestion and MALDI spotting. The process is performed without any human intervention, avoiding common contamination sources such as keratins.

Ettan Dalt six, IPGphore , Spot picker and Typhoon scanner TM9400 (GE Healthcare): these instruments allow performing the whole workflow of 2D-PAGE/2D-DIGE analysis.

Probot fraction collector and MALDI spotter (Dionex) is dedicated to LC-MALDI applications and can be hyphenated with all the LC systems available in the facility.

Gelfree 8100 (Expedeon) allows separating proteins according to their apparent molecular weight and recovering them in solution. It represents therefore an interesting prefractionnation step before 1D/2D UPLC analyses but requires a few hundreds micrograms of starting material.

ImagePrep (Bruker) is used to spray MALDI matrix on various samples such as tissue slices for MS molecular Imaging.

SunCollect (SunChrom) allows spraying solutions on tissue slices with very homogeneous coverage. The solution applied on the slice can be enzyme (like trypsin) for on-slice digestion or MALDI matrix for MS molecular Imaging.

ASE 200 Accelerated Solvent Extractor by DIONEX Benelux.

PowerPrep I & II by F.M.S.

Solid Phase Extraction (SPE) : Flash Vac-10 Vacuum Manifold by I.S.T

Other

hreels VGS-EELS 400: High resolution electron energy loss spectrometer. 0 to 120° detection. 0.002 V accuracy.

ISEELS: 2000 V ion shell electron energy loss spectrometer with position sensitive detector.

Uvikon XS: UV-Vis spectrophotometer (190 – 1100 nm)

Jobin Yvon CD6: Circular dichroïsm spectrophotometer

Softwares

Proteomics: Proteolynx Global Server with expression, Mascot server, Decyder, ProteinScape

Molecular modeling: Gaussian 03, Gaussian 09, Hyperchem 8, Spartan 04, NWCHEM, pcgamess, amber 9.

MS Imaging : FlexImaging, SCiLS.

The instruments were financed by

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