Purpose

The Preparative Biochemistry Unit provides equipment and methods to produce large quantities (several tens of mg) of pure and homogeneous proteins for functional and structural studies. Experiments can be performed by the staff members of the technical unit or by the users themselves. The facility is also equipped with a Liberty One CEM peptide synthesizer.

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Services

• Protein production projects by staff members (from a gene or recombinant plasmid): cloning into an expression plasmid, expression and solubility control, bacterial expression (and baculovirus system and mammalian cells in collaboration with the AniRA-AGC core facility), purification by chromatography or other system, quantification and quality control on circular dichroism biophysical equipment and diffusion light scattering.
• Antibody purification
• Peptide synthesis: up to 70 residues with a wide scale range between 0.05 and 5 mmol, with various post-translational modifications (biotinylation, chromophore binding 13C- and 15N-labeling, cyclization…). For small proteins, peptide synthesis can be a cost-effective solution to produce about 10 mg.
• Open access : Biochemistry/biophysics equipment and consumables for independent use, assistance and training in the use of the equipment.

Equipment

• 3 purification systems FPLC : AKTA PURE, AKTA Purifier (GE Healthcare), NGC (BioRad)
• 1 BioSprint 15 Robot (Qiagen) : mini purification of proteins on magnetic beads for expression test, solubility tests and purification tests (up to 15 tests in parallel).
• 1 Typhoon fluo-phosphoimager GE Healthcare : scanner laser, for detection and quantification of protein, nucleic acids and labelled molecules (radioactivity, fluorescence)
• 1 Caméra CCD Fusion Fx Vilber Lourmat : pictures of gels, blots
• Incubators Multitron Infors (from 15 to 40°C).
• 1 Nanodrop spectrophotometer for proteins and nucleic acids quantification
• 1 peptide synthesizer, Liberty One (CEM) with 2 HPLC (Agilent and Thermo).
• 1 DLS Diffusion Light Scattering Malvern NanoS, for homogeneity control of the purified proteins
• 1 circular dichroïsm spectrometer Chirascan (Applied Photophysics) for folding analysis of proteins

Training

1. Training before using the facility’s equipment is required, free of charge and must be validated before open access booking.

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fees

1. Information on user fees is available on intranet (contributors and academic team) or upon demand.

Documentation

1. Fiche préparation des échantillons
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Links

1. Réservation des équipements de biochime préparative (pour les utilisateurs déclarés – accès sécurisé)

2. How to find us  (intranet)