PICT
Plateforme Intégrée de Criblage de Toulouse
Integrated Screening Plateform of Toulouse
NAVIGATIOn
| CRYSTALLOGRAPHY | NMR | BIOPHYSICS | CHEMICAL SYNTHESIS |
| BIOINFORMATICS | ENZYME SCREENING |
Structural biology
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The macromolecular crystallography facility
The facility possesses all the necessary equipment to overcome the different steps leading to the crystal structure of any purified target. This includes:
- Nano-dispensing crystallization robot: Innovadyne Nanodrop ExtY.
- System for visualization and management of crystallization trials: TriTek CrystalPro XS.
- Fully automated storage and imaging system: Formulatrix Rock Imager 1000 equipped with visible and UV Optics, to be installed soon.
- Thermoregulated rooms at 4, 12 and 20 °C for the setting up of automated and manual crystallization trials.
- In-house X-ray data collection and processing equipment: a 4-circle diffractometer with CCD detector (XCalibur Nova, Oxford Diffraction/Agilent), a diffractometer specifically designed for the identification and analysis of protein crystals within multi-well crystallization plates (PX Scanner, Oxford Diffraction/Agilent).
- Binocular microscopes, cryogenic containers for storage and shipping of crystals.
The facility has access through individual or Block Allocation Group (BAG) proposals to several French and European synchrotrons (ESRF, Grenoble; SOLEIL, Gif/Yvette; ALBA, Barcelona; DESY, Hamburg).
The NMR facility
The facility contains three high-field state-of-the-art NMR spectrometers for data acquisition and computers for NMR analyses. The instrumentation presently available includes four-channel Bruker Avance 500, 600, and 700 MHz NMR spectrometers, devoted to high-resolution liquid-state and solid-state NMR. This equipment associated with various probe heads offers a large range of capabilities for biochemical analyses and structural biology.
- Bruker Avance 600MHz equipped with cryoprobe TCi (1H, 13C, 15N) is dedicated to SAR by NMR and structural biology, in solution exclusively. BEST is a liquid sample handler which enables to work with 96 deepwell plates. The CryoFIT is a Bruker accessory dedicated to cryoprobes which enables flux analysis by NMR. Together, these accessories allow analyzing wide series of samples in automated mode with the sensitivity provided by the cryoprobe. This equipment is therefore dedicated to the screening of ligands by NMR.
- Bruker Avance 700MHz equipped with QCi probe (1H, 13C, 15N, 31P) is dedicated to structural biology of macromolecules (protein and nucleic acid-protein interaction).
- Bruker Avance 500MHz equipped with TXi probe (1H, 13C, 15N).
The 500 and 700 MHz are versatile spectrometers that can be used for both liquid and solid samples (heterogeneous samples, amyloid fibrils, tissues, in-cell NMR, etc) depending on the probe installed. For that purpose, they are equipped with static and MAS probes with various spinning rates, from 12 KHz (4 mm O.D. rotor) to 60 KHz (1.3mm O.D. rotor) with two channels (X: broadband) to three channels (1H, 13C, 15N).
biophysical facility 
The equipment of the biophysical facility includes variety of instruments, which allow to probe the homogeneity and thermostability of the samples and to screen or characterize ligand-protein interactions using complementary approaches.
- Differential Scanning Fluorimetry (DSF): Biorad CFX96’s real time quantitative PCR detection for thermofluor.
- Circular Dischroism spectrometer (CD): Jobin-Yvon CD6 Circular Dichroism Spectrometer
- Micro-Isothermal Titration Calorimetry (ITC): Microcal/GE Healthcare iTC200 system
- Dynamic Light Scattering (DLS): Wyatt technology Dynapro Nanostar (DLS/SLS)
- FPLC with Size Exclusion Chromatography column (SEC) coupled with UV and RI detectors, DLS and Multi-Angle Light Scattering detectors (MALS): Wyatt technology DAWN HELEOS_8+ system
chemical facility 
Automated chemical synthesys
- Parallel synthetic robotic platform, SLT106 model – Chemspeed Technologies. Carries out automatic parallel chemical synthesis processes: reaction preparation, monitoring of the operating conditions and treatment of the reaction media. Our current implements allow us to perform up to 64 parallel reactions and to work between -20 and 145° C, under inert pressure and with reactive gas (up to 80 bars). (ITAV)
- Microwave assisted synthesis robotic platform, Swave model – Chemspeed Technologies. A fully automated platform to carry out any high throughput microwave synthesis (up to 144 reactions per sequence), from the preparation of reaction medium to tube setting and shipping to the microwave reactor. The Swave accessories and the SLT106 accessories are compatible, which makes the successive use of both platforms quick and easy. (ITAV)
Chemical library
- Liquid-handling robotic platform, Biomek NXP model - Beckman Coulter. It permits the management of the resulting chemical libraries. It is mainly operated through 96-Channel pipetting head and is compatible with DTX 880 microplate reader for performing fluorescence, luminescence and absorbance (visible-UV) measurements. (ITAV)
Chromatography equipments
- Acquity system (Waters): UHPLC reverse phase analyses. Analytical columns 5, 10 cm granulometry 1.7 µm available. (ICT, PICT)
- Pump 2545, injector/collector 2767, PDA2998, fractionlynx: analyses and purification of products under reverse or direct phase. Different analytical and preparative columns available. (ICT)
- Picsolution SFC: analyses and purification of chiral and non-chiral compounds. Analytical and semiprep columns, chiral (OD, AD, OJ, A, IC), and non-chiral (Silica and 2 Ethyl Pyridine) available. (ICT)
- Micro MX Masslynx (Waters): MALDI-TOF. Resolution Reflectron Masse/Width Signal 10000. (PICT)
- Pump 600, inj 717, PDA 996: reverse phase analyses. C18, Phenyl, CN (5, 10, 15, 25 cm, diameter 3 and 4.6 mm), hydrophilic, hydrophobic and large pH phases available. (LSPCMIB)
- Alliance 2695, PDA 2995, ELSD_2420: reverse phase analytical, C8 C18 CN Phenyl, Water Buffer MeOH, CH3CN, automated system. C18, Phenyl, CN (5, 10, 15, 25 cm, diameter 3 and 4.6 mm), hydrophilic, hydrophobic and large pH phases available. (LSPCMIB)
- Alliance 2695, PDA 2995, UV 486, ELSD_2420: direct phase analyses, silica, chiral columns. (LSPCMIB)
- Prochrom 50 (50 mm column to fill with 400 g of phase), UV Collector: Purification on silica gel or reverse phase, from 100 mg to 10 g/injection, HPLC quality separation flow 100 mL/min. Silica and C18 (10 and 12 µm) available. (LSPCMIB)
- Flash chromatography systems, CombiFlash model from Serlabo. Each implement has four programmable purifiers in direct or reversed phase. The self-priming pumps enable isocratic or binary gradient analysis. The implements are fitted out with a UV detection system. (ITAV)
chemo-informatics facility 
Equipment of the chemo-informatics facility includes two clusters for computational tasks as well as GPU servers.
Clusters
- A Dell cluster: 28 PowerEdge R410 servers, each equipped with two quadri-core 64-bit Intel Nehalem processors operating at 2.53 GHz, with 16 MHz cache memory and 12 Gb RAM. In total, there are 56 CPUs and 224 cores. The cluster's file server has a total capacity of 2.1 Tb, and is composed of 5 hard disks of 600 Gb each. The disks work in SATA2 mode, at 10000 rpm, and are secured by the RAID 5 protection system.
- A HP cluster, acquired in 2004, is composed of 20 Proliant DL140 servers, each equipped with two mono-core 32-bit Intel Xeon processors operating at 3.06 GHz, with 512 kHz cache memory and 2 Gb RAM. In total, there are 40 CPUs/cores.
- The cluster's "storage server" has a total capacity of 0.7 Tb, and is composed of 5 hard disks of 150 Gb each. The disks work in ATA mode, at 7200 rpm, and are secured by the RAID 5 protection system.
Softwares
- Softwares for molecular dynamics: Several different software packages have been installed, among which the most relevant are: Amber v.9, Gromacs v.4, Desmond v.2.4.2.1 and LAMMPS.
- Softwares for virtual screening: The docking programs that are used most often on the PICT platform are: AutoDock 3, 4 and Vina, Rosetta Suite v.3.1, DOCK v.6
The enzyme screening facility 
- QPixII (Genetix) : Colony picker (bacteria or yeasts) enabling the inoculation and re-arraying of mini-cultures in microtiterplates (3000 clones / hour)
- K2 (KBiosystems) : Multi-application robot allowing colony picking (contrast, color and fluorescence detection), plate replication, gridding on membrane/gelose, and colony re-arraying. Equipped with hotels for microtiterplate storage and a large worktable, the K2 robot provides high-picking and working speeds and excellent user autonomy.
- Genesis RSP200 (Tecan) : An integrated liquid handling station equipped with a robotic arm, various incubators and a spectrophotometer, and dedicated to the phenotypic screening of variant librairies (up to 10000 clones a week).
- Biomek 2000 (Beckman) : Transfer of liquids in microtiterplates under sterile conditions (2000 clones/ hour).
- Kühner : Microplate incubator-shakers for cell growth and protein expression in liquid medium (30°C, up to 264 microplates).
- Infors : Microplate incubator-shakers for cell growth and protein expression in liquid medium (10-65°C, up to 96 microplates).
- High-throughput HPLC (Dionex, Agilent) : Enzymatic activity screening via HPLC (3000 clones a week).
- LC/MS (Dionex) : Enzymatic activity screening, mixed molecule identification, direct injection or coupling to liquid chromatography.Eclipse
- Fluorimeter (Varian) : Fluorescence spectroscopy, protein-ligand ibnteractions, thermostability study, cuvette or 96 well microtiter plates
- Circular dichroism (Jasco) : Assessment of the structural integrity of the recombinant proteins
- Protein chromatography system ÄKTAxpress (GE Healthcare) : Multistep purification, 4 systems controlled in parallel : simultaneous purification of up to 4 proteins.