CNRS - AIX MARSEILLE UNIV: UMR7257

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Structural Biology

Recombinant protein production in eukaryotic systems

Heads Christine KELLENBERGER, Vincent DELAUZUN

Our service facility offers the latest technologies for production of soluble, membrane-linked or membrane-embedded eukaryotic proteins in mammalian (HEK293, CHO, COS-7) or insect cells (S2, Sf9), which are able to process most of the posttranslational modifications (disulfide bridge, N- and O-glycosylation, phosphorylation…) important for protein structure or function. The feasibility study associates the specification needs of the applicant (protein amounts, tags for purification and detection…) and the expertise of the facility staff. The selected protocol is assayed, validated and then amplified for a quantitative production assay. The produced proteins are purified and characterized by the applicants.

Accessibility

Academic and industrial parties

How to apply ?

Dedicated equipment

  • 4 CO2 incubators
  • 3 temperature-controlled incubators
  • 4 laminar flow hoods
  • 1 epifluorescence microscope
  • 1 inverted microscope

Cost

  • Phase 1 (evaluation): based on the system used.
  • Phase 2 (production): based on the expression yields and protein
    amounts required.

Publications linked to the techniques used

  • Durocher Y, Loignon M, Process, vectors and engineered cell lines for enhanced large-scale transfection, National Research Council of Canada, WO2009/137911.
  • Aricescu AR, Lu W, Jones EY (2006) A time- and cost-efficient system for high-level protein production in mammalian cells. Acta Cryst D62, 1243–1250.
  • Yao F, Svensjö T, Winkler T, Lu M, Eriksson C, Eriksson E (1998) Tetracycline repressor, tetR, rather than the tetR-mammalian cell transcription factor fusion derivatives, regulates inducible gene expression in mammalian cells. Hum Gene Ther 9, 1939-1950.
  • Liepman AH, Wilkerson CG, Keegstra K (2005) Expression of cellulose synthase-like (Csl) genes in insect cells reveals that CslA family members encode mannan synthases. Proc Natl Acad Sci USA 102:2221-2226.
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