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Synthelis and ESRF collaborate to provide cutting-edge structural biology services. Synthelis offers development, production, and characterization of difficult-to-express proteins, while ESRF provides access to state-of-the-art synchrotron radiation facilities and expertise. This collaboration offers a comprehensive and streamlined workflow for protein structure determination.
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1/ Collaboration key players • Synthelis and ESRF SYNTHELIS / www.synthelis.com
Synthelis, a dynamic company • Transforming academic expertise into relevant services • Spin-off company from University Joseph Fourier Grenoble - Incorporated in January 2011 • Bringing service offer to industries : development, production & characterization of difficult to express proteins (membrane & cytotoxic proteins) Proprietary technology and know-how in cell-free protein expression SYNTHELIS / www.synthelis.com
Recognized expertise • Already validated expertise with well-established customers • 33 client portfolio • 93% of protein successfully expressed • 77% with proof of activity • > 80 proteins successfully produced to date SYNTHELIS / www.synthelis.com
Structural biology cluster – Grenoble area SYNTHELIS / www.synthelis.com
ESRF in brief • European Synchrotron Radiation Facility (ESRF) • Established in 1975 with the synchrotron project, capable of producing highly brilliant hard X-rays. Services • Beam time measurements • Experiment support by in-house experts • Industrial access • Simple and effective access to X-ray facilities and expertise • Specialized laboratory services • Technology transfer, instrument sales and licensing World reference for industrial and academic use of synchrotron radiation SYNTHELIS / www.synthelis.com
2/ Integrated offer • From risk analysis to MAD/SAD structure solving SYNTHELIS / www.synthelis.com
Crystal optimization • Overall process* 10 to 14 months 1 week 1 week 5 - 6 months 1 month 1–2 month(s) 1-2 month(s) 1 week 2 weeks If bad diffraction If no crystals 1-2 month(s) * Approximated timelines SYNTHELIS / www.synthelis.com
Risk analysis • Service offer • Sequence analysis and structure prediction tools • Recommendation on which or how many construct(s) to be tested • Reporting and scientific meeting for GO/NO GO decision Sequence alignement Timeline 1 week Disorder prediction (Source: Thesis from E.Poudevigne) SYNTHELIS / www.synthelis.com
Se-Met expression & expression upscale • Service offer • Two genetic construction design and production • Screening of cell-free expression conditions using Se-Met amino acids • Pre-purification by affinity • Western-Blot for identification and Coomassie Blue Staining gel for quantification • Expression scale up to produce mgs for characterization, purification and crystallogenesis. Timeline 5 - 6 months SYNTHELIS / www.synthelis.com
Stability and activity assessment (TSA) • Service offer • For the 2 constructions in different conditions • Thermal shift assays to assess the activity and the thermo-stability of the proteins • Reporting and scientific meeting for GO/NO GO decision Timeline 1 month TSA measurements SYNTHELIS / www.synthelis.com
Purification set-up & optimization • Service offer • Purification with pre-determined protocol and concentration of sample up to 10mg/ml if possible • Concentration and solubility tests • Reporting (Source: Thèse E.Poudevigne) Timeline 1-2 month(s) SYNTHELIS / www.synthelis.com
Crystallogenesis • Service offer • Crystallization set-up • Image Inspection/annotation • Screening of 1 protein samples up to 3 concentrations (one screening consisting of 5-6 crystallization plates +1 refinement plate). Preparation of cryogenic solutions for up to best 5 hits per screen. • Reporting Timeline 1-2 month(s) SYNTHELIS / www.synthelis.com
Biophysics options • Service offer • Size and shape of macromolecules • Stoichiometry of macromolecular assemblies • Secondary and tertiary structure of macromolecules • Thermodynamic stability of macromolecules • Properties kinetic and thermodynamic interactions Timeline 2-6 weeks depending on options SYNTHELIS / www.synthelis.com
Diffraction test • Service offer • Test of crystal hits on synchrotron beamline • Feedback to crystallisation if furtherrefinementrequired Timeline 1 week SYNTHELIS / www.synthelis.com
Crystal optimization • Service offer • Manual production of crystals for data collection with optimized conditions from (5). Production of cryo-conditions, mounting and freezing crystals. Maximum duration of three months. • Manual crystal optimization • Consumables • Reporting Timeline 1 – 2 month(s) SYNTHELIS / www.synthelis.com
Structure solving and refinement • Service offer • Protein structure determination – via molecular replacement or use of anomalous scattering • Subsequent refinement of structure • Reporting to level equivalent to PDB structure deposition Timeline 2 weeks for standard resolution SYNTHELIS / www.synthelis.com
3/ Why work together ? SYNTHELIS / www.synthelis.com
Benefits • Complementary expert teams • Proximity between the production site and the structure determination site • Single point-of-contact to access the whole value chain • Industry-driven service offer SYNTHELIS / www.synthelis.com
Alex Abadie 5 Avenue du Grand Sablon 38700 Grenoble - FRANCE Phone : +33 4 76 54 95 35 Mobile : +33 6 46 23 12 60 alex.abadie@synthelis.com www.synthelis.com SYNTHELIS / www.synthelis.com
