Bringing structural biology services through collaboration

SYNTHELIS / 2 1/ Collaboration key players Synthelis and ESRF

SYNTHELIS / 3 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 / 4 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 / 5 Structural biology cluster – Grenoble area

SYNTHELIS / 6 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 / 7 2/ Integrated offer From risk analysis to MAD/SAD structure solving

SYNTHELIS / 8 Overall process* Risk analysis Se-Met expression Expression upscale Stability and activity assessment (TSA) Purification set-up & optimization Cristallo- genesis Biophysics options Diffraction test Data collection MAD/SAD structure solving If no crystals Crystal optimization If bad diffraction 1 week5 - 6 months1 month1–2 month(s)1-2 month(s)1 week 2 weeks * Approximated timelines 1-2 month(s ) 10 to 14 months

SYNTHELIS / 9 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 Disorder prediction (Source: Thesis from E.Poudevigne) Timeline 1 week Sequence alignement

SYNTHELIS / 10 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 months

SYNTHELIS / 11 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 TSA measurements Timeline 1 month

SYNTHELIS / 12 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 / 13 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 / 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 / 15 Diffraction test Service offer  Test of crystal hits on synchrotron beamline  Feedback to crystallisation if further refinement required Timeline 1 week

SYNTHELIS / 16 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 / 17 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 / / Why work together ?

SYNTHELIS / 19 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

Your contact at SYNTHELIS : SYNTHELIS / 20 Alex Abadie 5 Avenue du Grand Sablon Grenoble - FRANCE Phone : Mobile :